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CFTR (Fundulus heteroclitus) in pSGX4 (His and Smt3-tagged bacterial expression vector)

TOP

Explanation of Terms


Gene: Gene Symbol:  CFTR
Gene Name:  cystic fibrosis transmembrane conductance regulator
Sequence             
Original Clone ID: 5767b2BSt1p1
Keyword: Partial Protein, NBD1 domain
Species: Fundulus heteroclitus
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 389-645 on full length protein AAC41271.1.
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.
FhCD00287230

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AVAILABLE
No restriction
Special MTA: PSI/Smt3/SUMO

 

Insert sequence: 687nts         Open reading frame : 1 to 687

ACCACCGTTGGCCTGGAACTGATCAACGTGAGCTCCTCCTGGGACGAAGGCGGTACCCCG
GTTCTGAAAAACATCAACCTGCGTCTGGAGGAGGGTATGATGCTGGCAGTGGCGGGCTCC
ACCGGTTCTGGTAAAAGCTCTCTGCTGATGATGATCCTGGGCGAGCTGGTCCCATCCGAG
GGTAAAATCCGTCATAGCGGCCGTATTAGCTATAGCCCGCAGACTAGCTGGATTATGCCA
GGTACCATTCGCGATAACATCCTGTTTGGCCTGACCTACGATGAATTCCGTTACACGAGC
ATTATTCGCGCTTGCCAGCTGGAAGAGGACCTGGATCTGCTGCCGGAGAAAGACAAAACC
GCGATTGCGGAAGGTGGTGTCACCCTGTCTGGTGGCCAGCGTGCTCGCATTTGTCTGGCT
CGTGCAGTGTACAAGGATGCTGACCTGTATCTGCTGGACGCTCCGTTTACCCACCTGGAT
ATCGCTACTGAAAAAGAAATCTTCGACAAATGTGTCTGTAAACTGATTGCATCCAAAACC
CGTGTAGTCGTGACTTCTAAACTGGAACACCTGAAACGTGCCGATCGTATTCTGCTGCTG
CACAACGGCGACTGTTACTTCTATGGCACCTTCTCTGAGCTGCAAGCGCAGCGCCCGGAC
TTCAGCTCCCTGCTGCTGGGCTGATAA


Coding Sequence Details
Insert Sequence Verified?: Y Verification Method: End read sequencing of cDNA insert
Reference Sequence Annotations:
End on protein reference sequence 645
Start on protein reference sequence 389
Mutations:
del406-435, V436G

del@406,29

Discrepancy:
None
Others:
Domain NBD1
Termini 389-645
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: ProteinPurified: Protein_Purified
ProteinExpressed: Protein_Confirmed
SolubleProtein: Protein_Soluble
Find experimental protocols in 5767b
Recommended expression in: BL21(DE3)-Codon+RIL
       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
  • Protein
  • Clone
  • Experimental

Gene

Target GenBank: AAC41271.1

 

Protein

Domain : NBD1
Structural Annotation Wiki : TOPSAN

Clone

Cloning Information : 473981
HIP Master Clone ID : 395487
Original Clone ID : 5767b2BSt1p1

Experimental

TargetTrack - Experimental data : 5767b