LOCUS       40924_41335        3144 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pStartK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3144)
  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 3144)
  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 3144)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3144)
  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..3144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..101
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             256..558
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRIVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(590..689)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     rep_origin      822..1367
                     /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."
     CDS             1962..2774
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      2891..2977
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3069..3096
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca
       61 atgctttttt ataatgccaa ctttgtacaa aaaagcaggc tttaaaggaa ccaattcagt
      121 cgactggatc cggtaccgaa ttcgcttact aaaagccaga taacagtatg cgtatttgcg
      181 cgctgatttt tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag
      241 aggtgtgctt ctagaatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt
      301 ctgtttgtgg atgtacagag tgatattatt gacacgcccg ggcgacggat agtgatcccc
      361 ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat
      421 atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt
      481 atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac
      541 ctgatgttct ggggaatata gaattcgcgg ccgcactcga gatatctaga cccagctttc
      601 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
      661 actatcagtc aaaataaaat cattatttgc catccagctg cagctgtcaa acatgagaat
      721 tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
      781 gaaatctaga aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
      841 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
      901 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa
      961 cttgtccttt cagtttagcc ttaaccggcg catgacttca agactaactc ctctaaatca
     1021 attaccagtg gctgctgcca gtggtgcttt tgcatgtctt tccgggttgg actcaagacg
     1081 atagttaccg gataaggcgc agcggtcgga ctgaacgggg ggttcgtgca tacagtccag
     1141 cttggagcga actgcctacc cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc
     1201 ataacagcgg aatgacaccg gtaaaccgaa aggcaggaac aggagagcgc acgagggagc
     1261 cgccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac cactgatttg
     1321 agcgtcagat ttcgtgatgc ttgtcagggg ggcggagcct atggaaaaac ggctttgccg
     1381 cggccctctc acttccctgt taagtatctt cctggcatct tccaggaaat ctccgccccg
     1441 ttcgtaagcc atttccgctc gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga
     1501 ggaagcggaa tatatcctgt atcacatatt ctgctgacgc accggtgcag ccttttttct
     1561 cctgccacat gaagcacttc actgacaccc tcatcagtgc caacatagta agccagtata
     1621 cactccgcta gcgctgaggt ctgcctcgtg aagaaggtgt tgctgactca taccaggcct
     1681 gaatcgcccc atcatccagc cagaaagtga gggagccacg gttgatgaga gctttgttgt
     1741 aggtggacca gttggtgatt ttgaactttt gctttgccac ggaacggtct gcgttgtcgg
     1801 gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa caaagccacg
     1861 ttgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     1921 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     1981 aaacgtcttg ctcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     2041 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc
     2101 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2161 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2221 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat
     2281 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2341 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     2401 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     2461 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc
     2521 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     2581 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     2641 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     2701 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     2761 tcgatgagtt tttctaatca gacatgttct ttcctgcgtt atcccctgat tctgtggata
     2821 accgtattac cgctagcatg gatctcgggg acgtctaact actaagcgag agtagggaac
     2881 tgccaggcat caaataaaac gaaaggctca gtcggaagac tgggcctttc gttttatctg
     2941 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     3001 tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     3061 aactaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttcctgt
     3121 tagttagtta cttaagctcg ggcc
//
