LOCUS       V002121                 8491 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002121
VERSION     V002121
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8491)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Protoc"; date: "2008"; volume: "3"; issue: "6"; pages: "1056-1076"
            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"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..858
                     /label="AmpR"
                     /note="beta-lactamase"
     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"
     CDS             2361..3488
                     /gene="TK"
                     /label="Thymidine kinase"
                     /note="Thymidine kinase from Human herpesvirus 2 (strain
                     333). Accession#: P04407"
     protein_bind    4253..4377
                     /label="attR1"
                     /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..5142
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             5487..5789
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(5833..5957)
                     /label="attR2"
                     /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"
     5'UTR           6298..6337
                     /label="HSV TK 5'-UTR"
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             6355..7482
                     /label="HSV TK"
                     /note="herpes simplex virus thymidine kinase"
     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
                     /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 cgcgtttaaa
     1981 cggccctcga ctctagtcga gcagtgtggt tttcaagagg aagcaaaaag cctctccacc
     2041 caggcctgga atgtttccac ccaatgtcga gcagtgtggt tttgcaagag gaagcaaaaa
     2101 gcctctccac ccaggcctgg aatgtttcca cccaatgtcg agcaaacccc gcccagcgtc
     2161 ttgtcattgg cgaattcgaa cacgcagatg cagtctgggc ggcgcggccc gaggtccact
     2221 tcgcatatta aggtgacgcg cgtggcctcg aacagcgagc gaccctgcag cgacccgctc
     2281 atcagcgtca gcagcgttcc acaaatcctg gtggcgttga actcccgcac ctctcgggcg
     2341 aacgccttgt agaagcgggt atggcttctc acgccggcca acagcacgcg cctgcgttcg
     2401 gtcaggctgc tcgtgcgagc gggcctaccg acggccgcgc ggcgtcccgt cctagccatc
     2461 gccagggggc ctccgaagcc cgcggggatc cggagctgcc cacgctgctg cgggtttata
     2521 tagacggacc ccacggggtg gggaagacca ccacctccgc gcagctgatg gaggccctgg
     2581 ggccgcgcga caatatcgtc tacgtccccg agccgatgac ttactggcag gtgctggggg
     2641 cctccgagac cctgacgaac atctacaaca cgcagcaccg tctggaccgc ggcgagatat
     2701 cggccgggga ggcggcggtg gtaatgacca gcgcccagat aacaatgagc acgccttatg
     2761 cggcgacgga cgccgttttg gctcctcata tcggggggga ggctgtgggc ccgcaagccc
     2821 cgcccccggc cctcaccctt gttttcgacc ggcaccctat cgcctccctg ctgtgctacc
     2881 cggccgcgcg gtacctcatg ggaagcatga ccccccaggc cgtgttggcg ttcgtggccc
     2941 tcatgccccc gaccgcgccc ggcacgaacc tggtcctggg tgtccttccg gaggccgaac
     3001 acgccgaccg cctggccaga cgccaacgcc cgggcgagcg gcttgacctg gccatgctgt
     3061 ccgccattcg ccgtgtctac gatctactcg ccaacacggt gcggtacctg cagcgcggcg
     3121 ggaggtggcg ggaggactgg ggccggctga cgggggtcgc cgcggcgacc ccgcgccccg
     3181 accccgagga cggcgcgggg tctctgcccc gcatcgagga cacgctgttt gccctgttcc
     3241 gcgttcccga gctgctggcc cccaacgggg acttgtacca catttttgcc tgggtcttgg
     3301 acgtcttggc cgaccgcctc cttccgatgc atctatttgt cctggattac gatcagtcgc
     3361 ccgtcgggtg tcgagacgcc ctgttgcgcc tcaccgccgg gatgatccca acccgcgtca
     3421 caaccgccgg gtccatcgcc gagatacgcg acctggcgcg cacgtttgcc cgcgaggtgg
     3481 ggggagttta gttcaaacac ggaagcccga acggaaggcc tcccggcgat gacggcaata
     3541 aaagaacaga ataaaaggca ttgttgtcgt gtggtgtgtc cataagcgcg ggggttcggg
     3601 gccagggctg gcaccgtatc agcaccccac cgaaaaacgg agcgggccga tccgtccttg
     3661 ttttcggtct ggtactccct ttgtgctttt accctcaccc caccccatcc tttggcccgc
     3721 gcttacggca acaaagggcc tccgatagcc tccgaggtgc ggagcctctt tgggccgtgg
     3781 gtacggacac ccccccatct gcggactggc agccgggacg gacgaccatg ggccccggtc
     3841 tgtgggtggt gatgggggtc ctggtgggcg ttgccggggg ccatgacacg tactggacgg
     3901 agcaaatcga cccgtggttt ttgcacggtc tggggttggc ccgcacgtac tggcgcgaca
     3961 caaacaccgg gcgtctgtgg ttgcccaaca cccccgacgc cagcgacccc cagcgcggac
     4021 gcttggcgcc cccgggcgaa ctcaacctga ctacggcatc cgtgcccatg cttcggtggt
     4081 acgccgagcg cttttgtttc gtgttggtca ccacggccga gtttcctcgg gaccccgggc
     4141 agctgcttta catcccaaag acctatctgc tcggccggcc tcggaacgcg agcctgcccg
     4201 gaagatcccc gggtaccgag ctcgaattca tcgtcaccat cacctcgaat caacaagttt
     4261 gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
     4321 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggcg
     4381 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     4441 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     4501 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     4561 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     4621 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     4681 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     4741 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     4801 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     4861 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     4921 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     4981 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     5041 ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt
     5101 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaagatc tggatccggc
     5161 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     5221 tatactgata tgtatacccg aagtatgtca aaaagaggtg tgctatgaag cagcgtatta
     5281 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     5341 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     5401 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     5461 tgctgacgag aacagggact ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     5521 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     5581 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     5641 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     5701 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     5761 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     5821 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     5881 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     5941 ttcttgtaca aagtggttga ttcgaggctg ctaacaaatc gagtcgagca tcgagcagtg
     6001 tggttttcaa gaggaagcaa aaagcctctc cacccaggcc tggaatgttt ccacccaatg
     6061 tcgagcagtg tggttttgca agaggaagca aaaagcctct ccacccaggc ctggaatgtt
     6121 tccacccaat gtcgagcaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca
     6181 gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc
     6241 ctcgaacacc gagcgaccct gcagcgaccc gcttaacagc gtcaacagcg tgccgcagat
     6301 cttggtggcg tgaaactccc gcacctcttc ggccagcgcc ttgtagaagc gcgtatggct
     6361 tcgtaccccg gccatcaaca cgcgtctgcg ttcgaccagg ctgcgcgttc tcgcggccat
     6421 agcaaccgac gtacggcgtt gcgccctcgc cggcagcaag aagccacgga agtccgcccg
     6481 gagcagaaaa tgcccacgct actgcgggtt tatatagacg gtccccacgg gatggggaaa
     6541 accaccacca cgcaactgct ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc
     6601 gagccgatga cttactggcg ggtgctgggg gcttccgaga caatcgcgaa catctacacc
     6661 acacaacacc gcctcgacca gggtgagata tcggccgggg acgcggcggt ggtaatgaca
     6721 agcgcccaga taacaatggg catgccttat gccgtgaccg acgccgttct ggctcctcat
     6781 atcggggggg aggctgggag ctcacatgcc ccgcccccgg ccctcaccct catcttcgac
     6841 cgccatccca tcgccgccct cctgtgctac ccggccgcgc ggtaccttat gggcagcatg
     6901 accccccagg ccgtgctggc gttcgtggcc ctcatcccgc cgaccttgcc cggcaccaac
     6961 atcgtgcttg gggcccttcc ggaggacaga cacatcgacc gcctggccaa acgccagcgc
     7021 cccggcgagc ggctggacct ggctatgctg gctgcgattc gccgcgttta cgggctactt
     7081 gccaatacgg tgcggtatct gcagtgcggc gggtcgtggc gggaggactg gggacagctt
     7141 tcggggacgg ccgtgccgcc ccagggtgcc gagccccaga gcaacgcggg cccacgaccc
     7201 catatcgggg acacgttatt taccctgttt cgggcccccg agttgctggc ccccaacggc
     7261 gacctgtata acgtgtttgc ctgggccttg gacgtcttgg ccaaacgcct ccgttccatg
     7321 cacgtcttta tcctggatta cgaccaatcg cccgccggct gccgggacgc cctgctgcaa
     7381 cttacctccg ggatggtcca gacccacgtc accacccccg gctccatacc gacgatatgc
     7441 gacctggcgc gcacgtttgc ccgggagatg ggggaggcta actgaaacac ggaaggagac
     7501 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg
     7561 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     7621 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     7681 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     7741 ccactggccc cgtgggttag ggacggggtc ccccatgggg aatggtttat ggttcgtggg
     7801 ggttattatt ttgggcgttg cgtggggtca ggtccacgac ccaagcttgg ctgcaggtcg
     7861 agctcgcgaa agcttggcac tggccgtcgt tttggcactg gccgtcgttt tacaacgtcg
     7921 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     7981 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     8041 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     8101 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     8161 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     8221 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     8281 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     8341 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     8401 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     8461 aatgcttcaa taatattgaa aaaggaagag t
//
