LOCUS       40924_46648        6355 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWSTK6, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6355)
  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 6355)
  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 6355)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6355)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Protoc"; date: "2008"; volume: "3"; issue: "6"; pages: "1056-1076"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-FEB-2008) Howard Hughes Medical Institute, University of Utah, 
            15N 2030E, Salt Lake City, UT 84112, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..858
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1026..1570
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1854..1872
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2117..2241
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2266..2296
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2350..3006
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3351..3653
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3697..3821)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4078..4140
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     5'UTR           4162..4201
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             4219..5346
                     /codon_start=1
                     /label=HSV TK
                     /note="herpes simplex virus thymidine kinase"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    5391..5439
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(5761..5777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6251..6355
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagacccctt aataagatga
     1021 tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc
     1081 ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg aggtaactgg
     1141 cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc ggcgcatgac
     1201 ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg cttttgcatg
     1261 tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cggactgaac
     1321 ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac tgagtgtcag
     1381 gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac cgaaaggcag
     1441 gaacaggaga gcgcacgagg gagccgccag gggaaacgcc tggtatcttt atagtcctgt
     1501 cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag
     1561 cctatggaaa aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca
     1621 tcttccagga aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc
     1681 gagcgtagcg agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga
     1741 cgcaccggtg cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag
     1801 tgccaacata gtaagccagt atacactccg ctagcaacct ggcttatcga aattaatacg
     1861 actcactata gggagaccgg cagatccggc gcgcctgcag gcgcgccact agttaattaa
     1921 tttaaatcga tgcgatcgct agcggccgcg tttgcttttg tttcgtgttg gtcaccacgg
     1981 ccgagtttcc tcgggacccc gggcagctgc tttacatccc aaagacctat ctgctcggcc
     2041 ggcctcggaa cgcgagcctg cccggaagat ccccgggtac cgagctcgaa ttcatcgtca
     2101 ccatcacctc gaatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat
     2161 aaatatcaat atattaaatt agattttgca taaaaaacag actacataat actgtaaaac
     2221 acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac actttatgct
     2281 tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc aggagctaag
     2341 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     2401 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     2461 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     2521 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     2581 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     2641 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     2701 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     2761 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     2821 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     2881 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat
     2941 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     3001 ggggcgtaaa gatctggatc cggcttacta aaagccagat aacagtatgc gtatttgcgc
     3061 gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat gtcaaaaaga
     3121 ggtgtgctat gaagcagcgt attacagtga cagttgacag cgacagctat cagttgctca
     3181 aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga atgaagcccg
     3241 tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg
     3301 gtttattgaa atgaacggct cttttgctga cgagaacagg gactggtgaa atgcagttta
     3361 aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta cagagtgata
     3421 ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt ctgctgtcag
     3481 ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga
     3541 tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg gctgatctca
     3601 gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga atataaatgt
     3661 caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat atgttgtgtt
     3721 ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt gatatttata
     3781 tcattttacg tttctcgttc agctttcttg tacaaagtgg ttgattcgag gctgctaaca
     3841 aatcgagtcg agcatcgagc agtgtggttt tcaagaggaa gcaaaaagcc tctccaccca
     3901 ggcctggaat gtttccaccc aatgtcgagc agtgtggttt tgcaagagga agcaaaaagc
     3961 ctctccaccc aggcctggaa tgtttccacc caatgtcgag caaaccccgc ccagcgtctt
     4021 gtcattggcg aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc
     4081 gcatattaag gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa
     4141 cagcgtcaac agcgtgccgc agatcttggt ggcgtgaaac tcccgcacct cttcggccag
     4201 cgccttgtag aagcgcgtat ggcttcgtac cccggccatc aacacgcgtc tgcgttcgac
     4261 caggctgcgc gttctcgcgg ccatagcaac cgacgtacgg cgttgcgccc tcgccggcag
     4321 caagaagcca cggaagtccg cccggagcag aaaatgccca cgctactgcg ggtttatata
     4381 gacggtcccc acgggatggg gaaaaccacc accacgcaac tgctggtggc cctgggttcg
     4441 cgcgacgata tcgtctacgt acccgagccg atgacttact ggcgggtgct gggggcttcc
     4501 gagacaatcg cgaacatcta caccacacaa caccgcctcg accagggtga gatatcggcc
     4561 ggggacgcgg cggtggtaat gacaagcgcc cagataacaa tgggcatgcc ttatgccgtg
     4621 accgacgccg ttctggctcc tcatatcggg ggggaggctg ggagctcaca tgccccgccc
     4681 ccggccctca ccctcatctt cgaccgccat cccatcgccg ccctcctgtg ctacccggcc
     4741 gcgcggtacc ttatgggcag catgaccccc caggccgtgc tggcgttcgt ggccctcatc
     4801 ccgccgacct tgcccggcac caacatcgtg cttggggccc ttccggagga cagacacatc
     4861 gaccgcctgg ccaaacgcca gcgccccggc gagcggctgg acctggctat gctggctgcg
     4921 attcgccgcg tttacgggct acttgccaat acggtgcggt atctgcagtg cggcgggtcg
     4981 tggcgggagg actggggaca gctttcgggg acggccgtgc cgccccaggg tgccgagccc
     5041 cagagcaacg cgggcccacg accccatatc ggggacacgt tatttaccct gtttcgggcc
     5101 cccgagttgc tggcccccaa cggcgacctg tataacgtgt ttgcctgggc cttggacgtc
     5161 ttggccaaac gcctccgttc catgcacgtc tttatcctgg attacgacca atcgcccgcc
     5221 ggctgccggg acgccctgct gcaacttacc tccgggatgg tccagaccca cgtcaccacc
     5281 cccggctcca taccgacgat atgcgacctg gcgcgcacgt ttgcccggga gatgggggag
     5341 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5401 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5461 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5521 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5581 tcggggcggc aggccctgcc atagccactg gccccgtggg ttagggacgg ggtcccccat
     5641 ggggaatggt ttatggttcg tgggggttat tattttgggc gttgcgtggg gtcaggtcca
     5701 cgacccaagc ttggctgcag gtcgagctcg cgaaagcttg gcactggccg tcgttttggc
     5761 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5821 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5881 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     5941 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     6001 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     6061 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     6121 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     6181 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     6241 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     6301 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagt
//
