LOCUS       Exported                8491 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWSTK2, complete sequence.
ACCESSION   EU530623
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8491)
  AUTHORS   Wu S, Ying G, Wu Q, Capecchi MR.
  TITLE     A protocol for constructing gene targeting vectors: generating 
            knockout mice for the cadherin family and beyond
  JOURNAL   Nat Protoc 3 (6), 1056-1076 (2008)
  PUBMED    18546598
REFERENCE   2  (bases 1 to 8491)
  AUTHORS   Wu S, Capecchi MR.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-FEB-2008) Howard Hughes Medical Institute, University 
            of Utah, 15N 2030E, Salt Lake City, UT 84112, USA
REFERENCE   3  (bases 1 to 8491)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8491
                     /organism="Cloning vector pWSTK2"
                     /mol_type="other DNA"
                     /db_xref="taxon:512281"
     CDS             1..861
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin resistance protein"
                     /label=bla
                     /protein_id="ACC91757.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1..861
                     /gene="bla"
                     /label=bla
     rep_origin      1006..1679
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"
     rep_origin      1032..1620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        1878..1896
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2361..3491
                     /codon_start=1
                     /product="thymidine kinase"
                     /label=thymidine kinase
                     /note="derived from Herpes simplex virus; HSV-tk"
                     /protein_id="ACC91758.1"
                     /translation="MASHAGQQHAPAFGQAARASGPTDGRAASRPSHRQGASEARGDPE
                     LPTLLRVYIDGPHGVGKTTTSAQLMEALGPRDNIVYVPEPMTYWQVLGASETLTNIYNT
                     QHRLDRGEISAGEAAVVMTSAQITMSTPYAATDAVLAPHIGGEAVGPQAPPPALTLVFD
                     RHPIASLLCYPAARYLMGSMTPQAVLAFVALMPPTAPGTNLVLGVLPEAEHADRLARRQ
                     RPGERLDLAMLSAIRRVYDLLANTVRYLQRGGRWREDWGRLTGVAAATPRPDPEDGAGS
                     LPRIEDTLFALFRVPELLAPNGDLYHIFAWVLDVLADRLLPMHLFVLDYDQSPVGCRDA
                     LLRLTAGMIPTRVTTAGSIAEIRDLARTFAREVGGV"
     misc_recomb     4253..4377
                     /note="attR1; target DNA sequence for site-specific Gateway
                     recombinases"
     protein_bind    4253..4377
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4402..4432
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4486..5145
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol resistance protein"
                     /label=cat
                     /protein_id="ACC91759.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            4486..5145
                     /gene="cat"
                     /label=cat
     CDS             5487..5792
                     /codon_start=1
                     /gene="ccdB"
                     /product="F-plasmid toxin"
                     /label=ccdB
                     /protein_id="ACC91760.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            5487..5792
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     5833..5957
                     /note="attR2; target DNA sequence for site-specific Gateway
                     recombinases"
     protein_bind    complement(5833..5957)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6214..6276
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     5UTR           6298..6337
                     /label=HSV TK 5-UTR
                     /note="5 untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             6355..7485
                     /codon_start=1
                     /product="thymidine kinase"
                     /label=thymidine kinase
                     /note="derived from Herpes simplex virus; HSV-tk"
                     /protein_id="ACC91761.1"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    7527..7575
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(7897..7913)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8387..8491
                     /gene="bla"
                     /label=AmpR promoter