LOCUS       40924_46643        6379 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWSTK3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6379)
  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 6379)
  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 6379)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6379)
  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..6379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..858
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1032..1620
                     /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"
     protein_bind    2141..2265
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2290..2320
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2374..3030
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             3375..3677
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(3721..3845)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4102..4164
                     /label=Mini-TK promoter
                     /note="minimal herpes simplex virus (HSV) thymidine kinase 
                     promoter"
     5'UTR           4186..4225
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             4243..5370
                     /codon_start=1
                     /label=HSV TK
                     /note="herpes simplex virus thymidine kinase"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    5415..5463
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(5785..5801)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6275..6379
                     /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 cgtattgacg 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 cctgtagcaa 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 cagaccccgt agaaaagatc
     1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1141 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1681 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     1741 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     1801 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcaggtta
     1861 acctggctta tcgaaattaa tacgactcac tatagggaga ccggcagatc cggcgcgcct
     1921 gcaggcgcgc cactagttaa ttaatttaaa tcgatgcgat cgctagcggc cgcgtttgct
     1981 tttgtttcgt gttggtcacc acggccgagt ttcctcggga ccccgggcag ctgctttaca
     2041 tcccaaagac ctatctgctc ggccggcctc ggaacgcgag cctgcccgga agatccccgg
     2101 gtaccgagct cgaattcatc gtcaccatca cctcgaatca acaagtttgt acaaaaaagc
     2161 tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt tgcataaaaa
     2221 acagactaca taatactgta aaacacaaca tatccagtca ctatggcggc cgcattaggc
     2281 accccaggct ttacacttta tgcttccggc tcgtataatg tgtggatttt gagttaggat
     2341 ccgtcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
     2401 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
     2461 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
     2521 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
     2581 catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
     2641 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
     2701 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
     2761 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
     2821 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
     2881 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
     2941 caggttcatc atgccgtctg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
     3001 cagtactgcg atgagtggca gggcggggcg taaagatctg gatccggctt actaaaagcc
     3061 agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg
     3121 tatacccgaa gtatgtcaaa aagaggtgtg ctatgaagca gcgtattaca gtgacagttg
     3181 acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag
     3241 cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca
     3301 ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa
     3361 cagggactgg tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc
     3421 tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc
     3481 tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata
     3541 tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta
     3601 tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc
     3661 tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac
     3721 catagtgact ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
     3781 taatttaata tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa
     3841 gtggttgatt cgaggctgct aacaaatcga gtcgagcatc gagcagtgtg gttttcaaga
     3901 ggaagcaaaa agcctctcca cccaggcctg gaatgtttcc acccaatgtc gagcagtgtg
     3961 gttttgcaag aggaagcaaa aagcctctcc acccaggcct ggaatgtttc cacccaatgt
     4021 cgagcaaacc ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg
     4081 gcggcgcggt cccaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga
     4141 gcgaccctgc agcgacccgc ttaacagcgt caacagcgtg ccgcagatct tggtggcgtg
     4201 aaactcccgc acctcttcgg ccagcgcctt gtagaagcgc gtatggcttc gtaccccggc
     4261 catcaacacg cgtctgcgtt cgaccaggct gcgcgttctc gcggccatag caaccgacgt
     4321 acggcgttgc gccctcgccg gcagcaagaa gccacggaag tccgcccgga gcagaaaatg
     4381 cccacgctac tgcgggttta tatagacggt ccccacggga tggggaaaac caccaccacg
     4441 caactgctgg tggccctggg ttcgcgcgac gatatcgtct acgtacccga gccgatgact
     4501 tactggcggg tgctgggggc ttccgagaca atcgcgaaca tctacaccac acaacaccgc
     4561 ctcgaccagg gtgagatatc ggccggggac gcggcggtgg taatgacaag cgcccagata
     4621 acaatgggca tgccttatgc cgtgaccgac gccgttctgg ctcctcatat cgggggggag
     4681 gctgggagct cacatgcccc gcccccggcc ctcaccctca tcttcgaccg ccatcccatc
     4741 gccgccctcc tgtgctaccc ggccgcgcgg taccttatgg gcagcatgac cccccaggcc
     4801 gtgctggcgt tcgtggccct catcccgccg accttgcccg gcaccaacat cgtgcttggg
     4861 gcccttccgg aggacagaca catcgaccgc ctggccaaac gccagcgccc cggcgagcgg
     4921 ctggacctgg ctatgctggc tgcgattcgc cgcgtttacg ggctacttgc caatacggtg
     4981 cggtatctgc agtgcggcgg gtcgtggcgg gaggactggg gacagctttc ggggacggcc
     5041 gtgccgcccc agggtgccga gccccagagc aacgcgggcc cacgacccca tatcggggac
     5101 acgttattta ccctgtttcg ggcccccgag ttgctggccc ccaacggcga cctgtataac
     5161 gtgtttgcct gggccttgga cgtcttggcc aaacgcctcc gttccatgca cgtctttatc
     5221 ctggattacg accaatcgcc cgccggctgc cgggacgccc tgctgcaact tacctccggg
     5281 atggtccaga cccacgtcac cacccccggc tccataccga cgatatgcga cctggcgcgc
     5341 acgtttgccc gggagatggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5401 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     5461 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5521 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5581 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc actggccccg
     5641 tgggttaggg acggggtccc ccatggggaa tggtttatgg ttcgtggggg ttattatttt
     5701 gggcgttgcg tggggtcagg tccacgaccc aagcttggct gcaggtcgag ctcgcgaaag
     5761 cttggcactg gccgtcgttt tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5821 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5881 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5941 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     6001 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     6061 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6121 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     6181 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     6241 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6301 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6361 atattgaaaa aggaagagt
//
