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CFTR (Homo sapiens) in pSGX4 (His and Smt3-tagged bacterial expression vector)

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Explanation of Terms


Gene: Gene Symbol:  CFTR
Gene Name:  cystic fibrosis transmembrane conductance regulator (ATP-binding cassette sub-family C, member 7)
Sequence             
Original Clone ID: 2935c471BSt2p1
Keyword: Partial Protein, NBD1 domain
Species: Homo sapiens
Type: cDNA
Vector Name: pSGX4               Format:  FUSION
Source: SGX Pharmaceuticals, Inc.
Description: This clone encodes the NBD1 domain with other intended mutations and corresponds to region 387-646 on full length protein NP_000483.3.
Comments: N-terminal 6-His and Smt3, cleavable, TOPO. Insert sequence of this clone was provided by clone submitter and was not fully sequence verified.
Mutation/
Discrepancy :
Yes / Unknown
Publications: None
Authors: SGX Pharmaceuticals, Inc.
HsCD00287382

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AVAILABLE
No restriction
Special MTA: PSI/Smt3/SUMO

 

Insert sequence: 783nts        
Open reading frame : 1 to 783
Reference Sequence Alignment

ACGACTACAGAAGTAGTGATGGAGAATGTAACAGCCTTCTGGGAGGAGGGATTTGGGGAA
TTATTAGAGAAAGCAAAACAAAACAATAACAATAGAAAAACTTCTAATGGTGATGACAGC
CTCTCCTTCAGTAATTTATCACTTCTTGGTACTCCTGTCCTGAAAGATATTAATTTCAAG
ATAGAAAGAGGACAGTTGTTGGCGGTTGCTGGATCCACTGGAGCAGGCAAGACTTCACTT
CTAATGATGATTATGGGAGAACTGGAGCCTTCAGAGGGTAAAATTAAGCACAGTGGAAGA
ATTTCATTCTGTTCTCAGTTTTCCTGGATTATGCCTGGCACCATTAAAGAAAATATCATC
TTTGGTGTTTCCTATGATGAATATAGATACAGAAGCGTCATCAAAGCATGCCAACTAGAA
GAAGACATCTCCAAGTTTGCAGAGAAAGACAATATAGTTCTTGGAGAAGGTGGAATCACA
CTGAGTGGAGGTCAACGAGCAAGAATTTCTTTAGCAAGAGCAGTATACAAAGATGCTGAT
TTGTATTTATTAGACTCTCCTTTTGGATACCTAGATGTTTTAACAGAAAAAGAAATATTT
GAAAGCTGTGTCTGTAAACTGATGGCTAACAAAACTAGGATTTTGGTCACTTCTAAAATG
GAACATTTAAAGAAAGCTGACAAAATATTAATTTTGCATGAAGGTAGCAGCTATTTTTAT
GGGACATTTTCAGAACTCCAAAATCTACAGCCAGACTTTAGCTCAAAACTCATGGGATGA
TAA


Coding Sequence Details
Insert Sequence Verified?: Y Verification Method: End read sequencing of cDNA insert
Reference Sequence Annotations:
Start on protein reference sequence 387
End on protein reference sequence 646
Mutations:
F409L, F429S, F433L

F409L; F429S; F433L

Discrepancy:
None
Others:
Termini 387-646
Domain NBD1
5' Linker Sequence:
ACAGAGAACAGATTGGTGGATCCCTT

3' Linker Sequence: None

 

 Recommended Growth Condition:

Distributed in bacterial strain : DH5-alpha T1 phage resistant
Antibiotic Selection: Host Type:  bacterial    Marker:  kanamycin
Bacterial Selection Condition:  50ug/mL kanamycin
Growth Condition:  Growth with the single antibiotic in LB at 37 degrees is recommended.
Comments:  Conditions for Gateway-type vectors in recombined (with insert) form and other kanamycin resistant vectors.
Protein Expression Results: ProteinExpressed: Protein_Confirmed
ProteinPurified: Protein_Purified
SolubleProtein: Tested_Not_Soluble
Find experimental protocols in 2935c
Recommended expression in: Not Applicable
       DyNA Vector Map

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Vector Name: pSGX4

pSGX4 in advanced viewed

Synonyms: pSGX-4, pSGX4_BS
Sequencing Primer: Forward:  T7
Reverse:  T7 terminator
Description: Bacterial (E. coli) expression vector with T7 promotor, adds N-terminal 6xHis and smt-3 tags, cleavage site; kanamycin resistance; TOPO cloning.
Comments: Modified from pET26b from Novagen. N-terminal 6xHis-tag followed by smt3 fusion protein. Smt3 fusion cloned into NheI and BamHI sites. TOPO adaptation at BamHI and XhoI sites.
Size (bp): 5611
Parent Vector: None
Empty Vector: None
Properties: TOPO Cloning, bacterial expression, in vitro transcription, with tag/fusion/marker
Author Name: New York SGX Research Center for Structural Genomics
Publications: None
Vector Map:         Vector Sequence:
Sequence:

 



Vector Features:

Type Name Description Start Position End Position
TOPO recognition site TOPO TOPO cloning site 5435 5444
bacterial origin ColE ColE1 bacterial origin of replication 1421 2103
gene fragment ROP ROP 2527 2703
promoter T7 T7 promoter 4984 5003
protease cleavage site ULP site ULP cleavage site (recognizes the tertiary structure of SUMO) 5140 5140
repressor protein gene LacI lacI cds 3515 4351
ribosome binding site RBS ribosome binding site (RBS) 5062 5068
selectable marker KanR kanamycin resistance gene 563 1375
tag Smt3 Smt3 tag 5140 5424
tag His 6xHis tag 5083 5100
trxn regulatory element lac operator lac operator 5003 5025
trxn termination sequence T7 term T7 termination sequence 5539 5585

 

  • Gene
  • Pathways
  • Protein
  • Clone
  • Experimental
  • Organism
  • Genome
  • Reagents

Gene

 

Gene Symbol : CFTR
Symbol Nomenclature : CFTR
SYNONYM : ABC35; ABCC7; CF; CFTR/MRP; MRP7; TNR-CFTR; dJ760C5.1
Designation : cAMP-dependent chloride channel|channel conductance-controlling ATPase|cystic fibrosis transmembrane conductance regulator
Full Nomenclature : cystic fibrosis transmembrane conductance regulator (ATP-binding cassette sub-family C, member 7)
GENEID : 1080
Locus Tag : tcag7.78
GenBank Accession : NP_000483.3
HGNC : 1884
MIM : 602421
Vega : OTTHUMG00000023076
Target GenBank: NP_000483.3

 

Reference Sequence Alignment

  

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATGCAGAGGTCGCCTCTGGAAAAGGCCAGCGTTGTCTCCAAACTTTTTTTCAGCTGGACC
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AGACCAATTTTGAGGAAAGGATACAGACAGCGCCTGGAATTGTCAGACATATACCAAATC
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CCTTCTGTTGATTCTGCTGACAATCTATCTGAAAAATTGGAAAGAGAATGGGATAGAGAG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CTGGCTTCAAAGAAAAATCCTAAACTCATTAATGCCCTTCGGCGATGTTTTTTCTGGAGA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TTTATGTTCTATGGAATCTTTTTATATTTAGGGGAAGTCACCAAAGCAGTACAGCCTCTC
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TTACTGGGAAGAATCATAGCTTCCTATGACCCGGATAACAAGGAGGAACGCTCTATCGCG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATTTATCTAGGCATAGGCTTATGCCTTCTCTTTATTGTGAGGACACTGCTCCTACACCCA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GCCATTTTTGGCCTTCATCACATTGGAATGCAGATGAGAATAGCTATGTTTAGTTTGATT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TATAAGAAGACTTTAAAGCTGTCAAGCCGTGTTCTAGATAAAATAAGTATTGGACAACTT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GTTAGTCTCCTTTCCAACAACCTGAACAAATTTGATGAAGGACTTGCATTGGCACATTTC
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GTGTGGATCGCTCCTTTGCAAGTGGCACTCCTCATGGGGCTAATCTGGGAGTTGTTACAG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GCGTCTGCCTTCTGTGGACTTGGTTTCCTGATAGTCCTTGCCCTTTTTCAGGCTGGGCTA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GGGAGAATGATGATGAAGTACAGAGATCAGAGAGCTGGGAAGATCAGTGAAAGACTTGTG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATTACCTCAGAAATGATTGAAAATATCCAATCTGTTAAGGCATACTGCTGGGAAGAAGCA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATGGAAAAAATGATTGAAAACTTAAGACAAACAGAACTGAAACTGACTCGGAAGGCAGCC
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TATGTGAGATACTTCAATAGCTCAGCCTTCTTCTTCTCAGGGTTCTTTGTGGTGTTTTTA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TCTGTGCTTCCCTATGCACTAATCAAAGGAATCATCCTCCGGAAAATATTCACCACCATC
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TCATTCTGCATTGTTCTGCGCATGGCGGTCACTCGGCAATTTCCCTGGGCTGTACAAACA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TGGTATGACTCTCTTGGAGCAATAAACAAAATACAGGATTTCTTACAAAAGCAAGAATAT
                                                                              

HsCD00287382      ---------------------ACGACTACAGAAGTAGTGATGGAGAATGTAACAGCCTTC
NM_000492.3       AAGACATTGGAATATAACTTAACGACTACAGAAGTAGTGATGGAGAATGTAACAGCCTTC
                                       ***************************************

HsCD00287382      TGGGAGGAGGGATTTGGGGAATTATTAGAGAAAGCAAAACAAAACAATAACAATAGAAAA
NM_000492.3       TGGGAGGAGGGATTTGGGGAATTATTTGAGAAAGCAAAACAAAACAATAACAATAGAAAA
                  **************************:*********************************

HsCD00287382      ACTTCTAATGGTGATGACAGCCTCTCCTTCAGTAATTTATCACTTCTTGGTACTCCTGTC
NM_000492.3       ACTTCTAATGGTGATGACAGCCTCTTCTTCAGTAATTTCTCACTTCTTGGTACTCCTGTC
                  ************************* ************.*********************

HsCD00287382      CTGAAAGATATTAATTTCAAGATAGAAAGAGGACAGTTGTTGGCGGTTGCTGGATCCACT
NM_000492.3       CTGAAAGATATTAATTTCAAGATAGAAAGAGGACAGTTGTTGGCGGTTGCTGGATCCACT
                  ************************************************************

HsCD00287382      GGAGCAGGCAAGACTTCACTTCTAATGATGATTATGGGAGAACTGGAGCCTTCAGAGGGT
NM_000492.3       GGAGCAGGCAAGACTTCACTTCTAATGGTGATTATGGGAGAACTGGAGCCTTCAGAGGGT
                  ***************************.********************************

HsCD00287382      AAAATTAAGCACAGTGGAAGAATTTCATTCTGTTCTCAGTTTTCCTGGATTATGCCTGGC
NM_000492.3       AAAATTAAGCACAGTGGAAGAATTTCATTCTGTTCTCAGTTTTCCTGGATTATGCCTGGC
                  ************************************************************

HsCD00287382      ACCATTAAAGAAAATATCATCTTTGGTGTTTCCTATGATGAATATAGATACAGAAGCGTC
NM_000492.3       ACCATTAAAGAAAATATCATCTTTGGTGTTTCCTATGATGAATATAGATACAGAAGCGTC
                  ************************************************************

HsCD00287382      ATCAAAGCATGCCAACTAGAAGAAGACATCTCCAAGTTTGCAGAGAAAGACAATATAGTT
NM_000492.3       ATCAAAGCATGCCAACTAGAAGAGGACATCTCCAAGTTTGCAGAGAAAGACAATATAGTT
                  ***********************.************************************

HsCD00287382      CTTGGAGAAGGTGGAATCACACTGAGTGGAGGTCAACGAGCAAGAATTTCTTTAGCAAGA
NM_000492.3       CTTGGAGAAGGTGGAATCACACTGAGTGGAGGTCAACGAGCAAGAATTTCTTTAGCAAGA
                  ************************************************************

HsCD00287382      GCAGTATACAAAGATGCTGATTTGTATTTATTAGACTCTCCTTTTGGATACCTAGATGTT
NM_000492.3       GCAGTATACAAAGATGCTGATTTGTATTTATTAGACTCTCCTTTTGGATACCTAGATGTT
                  ************************************************************

HsCD00287382      TTAACAGAAAAAGAAATATTTGAAAGCTGTGTCTGTAAACTGATGGCTAACAAAACTAGG
NM_000492.3       TTAACAGAAAAAGAAATATTTGAAAGCTGTGTCTGTAAACTGATGGCTAACAAAACTAGG
                  ************************************************************

HsCD00287382      ATTTTGGTCACTTCTAAAATGGAACATTTAAAGAAAGCTGACAAAATATTAATTTTGCAT
NM_000492.3       ATTTTGGTCACTTCTAAAATGGAACATTTAAAGAAAGCTGACAAAATATTAATTTTGCAT
                  ************************************************************

HsCD00287382      GAAGGTAGCAGCTATTTTTATGGGACATTTTCAGAACTCCAAAATCTACAGCCAGACTTT
NM_000492.3       GAAGGTAGCAGCTATTTTTATGGGACATTTTCAGAACTCCAAAATCTACAGCCAGACTTT
                  ************************************************************

HsCD00287382      AGCTCAAAACTCATGGGATGATAA------------------------------------
NM_000492.3       AGCTCAAAACTCATGGGATGTGATTCTTTCGACCAATTTAGTGCAGAAAGAAGAAATTCA
                  ********************: *:                                    

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATCCTAACTGAGACCTTACACCGTTTCTCATTAGAAGGAGATGCTCCTGTCTCCTGGACA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GAAACAAAAAAACAATCTTTTAAACAGACTGGAGAGTTTGGGGAAAAAAGGAAGAATTCT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATTCTCAATCCAATCAACTCTATACGAAAATTTTCCATTGTGCAAAAGACTCCCTTACAA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATGAATGGCATCGAAGAGGATTCTGATGAGCCTTTAGAGAGAAGGCTGTCCTTAGTACCA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GATTCTGAGCAGGGAGAGGCGATACTGCCTCGCATCAGCGTGATCAGCACTGGCCCCACG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CTTCAGGCACGAAGGAGGCAGTCTGTCCTGAACCTGATGACACACTCAGTTAACCAAGGT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CAGAACATTCACCGAAAGACAACAGCATCCACACGAAAAGTGTCACTGGCCCCTCAGGCA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AACTTGACTGAACTGGATATATATTCAAGAAGGTTATCTCAAGAAACTGGCTTGGAAATA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AGTGAAGAAATTAACGAAGAAGACTTAAAGGAGTGCTTTTTTGATGATATGGAGAGCATA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CCAGCAGTGACTACATGGAACACATACCTTCGATATATTACTGTCCACAAGAGCTTAATT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TTTGTGCTAATTTGGTGCTTAGTAATTTTTCTGGCAGAGGTGGCTGCTTCTTTGGTTGTG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CTGTGGCTCCTTGGAAACACTCCTCTTCAAGACAAAGGGAATAGTACTCATAGTAGAAAT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AACAGCTATGCAGTGATTATCACCAGCACCAGTTCGTATTATGTGTTTTACATTTACGTG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GGAGTAGCCGACACTTTGCTTGCTATGGGATTCTTCAGAGGTCTACCACTGGTGCATACT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CTAATCACAGTGTCGAAAATTTTACACCACAAAATGTTACATTCTGTTCTTCAAGCACCT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATGTCAACCCTCAACACGTTGAAAGCAGGTGGGATTCTTAATAGATTCTCCAAAGATATA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GCAATTTTGGATGACCTTCTGCCTCTTACCATATTTGACTTCATCCAGTTGTTATTAATT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GTGATTGGAGCTATAGCAGTTGTCGCAGTTTTACAACCCTACATCTTTGTTGCAACAGTG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CCAGTGATAGTGGCTTTTATTATGTTGAGAGCATATTTCCTCCAAACCTCACAGCAACTC
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AAACAACTGGAATCTGAAGGCAGGAGTCCAATTTTCACTCATCTTGTTACAAGCTTAAAA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GGACTATGGACACTTCGTGCCTTCGGACGGCAGCCTTACTTTGAAACTCTGTTCCACAAA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GCTCTGAATTTACATACTGCCAACTGGTTCTTGTACCTGTCAACACTGCGCTGGTTCCAA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATGAGAATAGAAATGATTTTTGTCATCTTCTTCATTGCTGTTACCTTCATTTCCATTTTA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ACAACAGGAGAAGGAGAAGGAAGAGTTGGTATTATCCTGACTTTAGCCATGAATATCATG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AGTACATTGCAGTGGGCTGTAAACTCCAGCATAGATGTGGATAGCTTGATGCGATCTGTG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AGCCGAGTCTTTAAGTTCATTGACATGCCAACAGAAGGTAAACCTACCAAGTCAACCAAA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CCATACAAGAATGGCCAACTCTCGAAAGTTATGATTATTGAGAATTCACACGTGAAGAAA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GATGACATCTGGCCCTCAGGGGGCCAAATGACTGTCAAAGATCTCACAGCAAAATACACA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GAAGGTGGAAATGCCATATTAGAGAACATTTCCTTCTCAATAAGTCCTGGCCAGAGGGTG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GGCCTCTTGGGAAGAACTGGATCAGGGAAGAGTACTTTGTTATCAGCTTTTTTGAGACTA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CTGAACACTGAAGGAGAAATCCAGATCGATGGTGTGTCTTGGGATTCAATAACTTTGCAA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CAGTGGAGGAAAGCCTTTGGAGTGATACCACAGAAAGTATTTATTTTTTCTGGAACATTT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AGAAAAAACTTGGATCCCTATGAACAGTGGAGTGATCAAGAAATATGGAAAGTTGCAGAT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GAGGTTGGGCTCAGATCTGTGATAGAACAGTTTCCTGGGAAGCTTGACTTTGTCCTTGTG
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GATGGGGGCTGTGTCCTAAGCCATGGCCACAAGCAGTTGATGTGCTTGGCTAGATCTGTT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       CTCAGTAAGGCGAAGATCTTGCTGCTTGATGAACCCAGTGCTCATTTGGATCCAGTAACA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       TACCAAATAATTAGAAGAACTCTAAAACAAGCATTTGCTGATTGCACAGTAATTCTCTGT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GAACACAGGATAGAAGCAATGCTGGAATGCCAACAATTTTTGGTCATAGAAGAGAACAAA
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       GTGCGGCAGTACGATTCCATCCAGAAACTGCTGAACGAGAGGAGCCTCTTCCGGCAAGCC
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       ATCAGCCCCTCCGACAGGGTGAAGCTCTTTCCCCACCGGAACTCAAGCAAGTGCAAGTCT
                                                                              

HsCD00287382      ------------------------------------------------------------
NM_000492.3       AAGCCCCAGATTGCTGCTCTGAAAGAGGAGACAGAAGAAGAGGTGCAAGATACAAGGCTT
                                                                              

HsCD00287382      ---
NM_000492.3       TAG
                     

Pathways

Reactome : ABC-family proteins mediated transport
Reactome : Transmembrane transport of small molecules

Protein

Domain : NBD1
Structural Annotation Wiki : TOPSAN
UniProt : P13569
HPRD : 03883

Clone

Cloning Information : 474133
HIP Master Clone ID : 395639
Original Clone ID : 2935c471BSt2p1

Experimental

TargetTrack - Experimental data : 2935c

Organism


TAX_ID : 9606
Species Specific ID: 1080

Genome

Chromosome : 7
Map Location : 7q31.2
Ensembl : ENSG00000001626

Reagents

Labome : CFTR-antibody