LOCUS       40924_33917        6938 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Chromosomal integration vector pOSIP-KP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6938)
  AUTHORS   St-Pierre F, Cui L, Priest DG, Endy D, Dodd IB, Shearwin KE.
  TITLE     One-step cloning and chromosomal integration of DNA
  JOURNAL   ACS Synth Biol 2 (9), 537-541 (2013)
  PUBMED    24050148
REFERENCE   2  (bases 1 to 6938)
  AUTHORS   St-Pierre F, Cui L, Priest DG, Endy D, Dodd IB, Shearwin KE.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-MAY-2013) Molecular and Biomedical Science, The 
            University of Adelaide, Molecular Life Sciences Building, Adelaide, 
            South Australia 5005, Australia
REFERENCE   3  (bases 1 to 6938)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6938)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol"; date: "2013"; volume: "2"; issue: "9"; pages: "537-541"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-MAY-2013) Molecular and Biomedical Science, The University of 
            Adelaide, Molecular Life Sciences Building, Adelaide, South 
            Australia 5005, Australia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             151..453
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     rep_origin      486..1074
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1134..1154
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1203..1230
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1279..1310)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     protein_bind    1387..1420
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(1430..2140)
                     /label=lambda repressor (ts)
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     RBS             2229..2237
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2372..3577
                     /label=phage phi-80 integrase
                     /note="integrase from phage phi-80"
     CDS             3757..4569
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4694..5082
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     protein_bind    5093..5140
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    5107..5140
                     /note="FRT; Flp recombinase target site"
     protein_bind    complement(5247..5564)
                     /label=phage Phi-80 attP
                     /note="attachment site of phage phi-80"
     terminator      complement(5653..5899)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     terminator      6219..6305
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6400..6486
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6581..6667
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(6765..6808)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gaattcgagc tcggtacccg gggatccgcg gccgcttcta gagggcttac taaaagccag
       61 ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata ctgatatgta
      121 tacccgaagt atgtcaaaaa gaggtatgct atgcagttta aggtttacac ctataaaaga
      181 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga
      241 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
      301 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
      361 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
      421 aaaaacgcca ttaacctgat gttctgggga atataataac ccgtagaaaa gatcaaaaga
      481 tcttcttgag atcctttttt tctgcgcgta atctgctact tgcaaacaaa aaaaccaccg
      541 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      601 ggctttagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
      661 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      721 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      781 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      841 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      901 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      961 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1021 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1081 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca cattactagt
     1141 agcggccgct gcagctcgag atgcatggcg cctaacctaa actgacaggc atcaaattaa
     1201 gcagaaggcc atcctgacgg atggcctttt tgcgtttcga acaattgaaa aaacctcgcg
     1261 ccttacctgt tgagtaatag tcaaaagcct ccggtcggag gcttttgact ttctgcttac
     1321 tgaatttcgg tggtgccgtt aattaaccgg tgggccctca tgataataat ggtttcttag
     1381 acgtccgaag ttcctattct ctagaaagta taggaacttc cctaggtcag ccaaacgtct
     1441 cttcaggcca ctgactagcg ataactttcc ccacaacgga acaactctca ttgcatggga
     1501 tcattgggta ctgtgggttt agtggttgta aaaacacctg accgctatcc ctgatcagtt
     1561 tcttgaaggt aaactcatca cccccaagtc tggctatgca gaaatcacct ggctcaacag
     1621 cctgctcagg gtcaacgaga attaacattc cgtcaggaaa gcttggcttg gagcctgttg
     1681 gtgcggtcat ggaattacct tcaacctcaa gccagaatgc agaatcactg gcttttttgg
     1741 ttgtgcttac ccatctctcc gcatcacctt tggtaaaggt tctaagctta ggtgagaaca
     1801 tccctgcctg aacatgagaa aaaacagggt actcatactc acttctaagt gacggctgca
     1861 tactaaccgc ttcatacatc tcgtagattt ctctggcgat tgaagggcta aattcttcaa
     1921 cgctaacttt gagaattttt gtaagcaatg cggcgttata agcatttaat gcattgatgc
     1981 cattaaataa agcaccaacg cctgactgcc ccatccccat cttgtctgcg acagattcct
     2041 gggataagcc aagttcattt ttcttttttt cataaattgc tttaaggcga cgtgcgtcct
     2101 caagctgctc ttgtgttaat ggtttctttt ttgtgctcat acgttaaatc tatcaccgca
     2161 agggataaat atctaacacc gtgcgtgttg actattttac ctctggcggt gataatggtt
     2221 gcatgtacta aggaggttgt atacgcgtta gcaaccctta tccagcaggc tttcaaggat
     2281 gtaaaccata acactctgcg aactagtgtt acattgcgtg tagctttgag tgggcaactt
     2341 tgtgtacact tttgtgtacc caaaaacaaa aatgtgtacc cattcaatga tcaccgacac
     2401 aaagctcagg aaggcgctcg gcaagaaaag agatgatatc gagattattt ctgattcgca
     2461 cgggctcaac gccagaatca gccaggccgg aaaaatatca tttttctatc ggtatcgctg
     2521 ggccggtaaa gcggtaaaac tcaatgttgg tgattatcct gcaatgagta tcacccaggc
     2581 aagagagcgt cgccaacaat tcagaaactg gttaactgag ggactggatc cgcgagagca
     2641 ggtgaagctg gataagcaga cccgacagga agcgatgtcc gttgccgaag cgttcaatta
     2701 ctggattgaa aggcactgta tcgctaacgg gctagttaaa gtcgattact atcgccaggt
     2761 gtttgagaaa catatcgccg aaccgatgaa gaatgtcaaa gtcgataaca cagcgaaaat
     2821 gcactggatc aacgtcttcg attctataga aagcagggtg atggctcatt acatgctttc
     2881 gctgtgcaaa cgggcgttta ggttctgcgt taacagaagt gtgatcgcct caaacccgct
     2941 cgagggatta ctgccatctg atgtcgggca aaagcctaaa aagagaactc gcaggatgga
     3001 cgatgacgat ctgcgcaaaa tctatcagtg gttgaaaagc catatgtcga tagagtccgt
     3061 tttcctggtg aaatttatta tgcttaccgg atgccgtacg gctgagattc gacttagtga
     3121 gagatcatgg tttcgattgg atgataatga gtgggtcgtg cctgcgggca gttataaaac
     3181 tcgggtacat attagaaggg gactctcaga cgccgccgtt aacctggtca gaaatcacct
     3241 caagaaaata aacaccaatc acctggtgac ttcacaacgt aaaattgatg gcgggatcaa
     3301 agattcgccc gttcattcac ctgtggcatc caattacgcc cgttctattt ggaatggaac
     3361 aggtatggca gagtggtcgc ttcatgatat gaggcggacg atagccacaa atctctctga
     3421 gttaggttgc ccgccgcacg taattgaaaa gctgctcggg catcagatgg tgggggttat
     3481 ggcgcattac aaccttcatg actatatcga tgatcagaaa cactggctcc gcgtttggca
     3541 gagccatctt gaagagatca tcggagagcc cttcagttaa tttatcttct ttttatcctc
     3601 ccactctttg attgccatgg gtatggacag ttttcccttt gatatgtaac gcacgttgtg
     3661 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     3721 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     3781 tcttgctccc gtccgcgctt aaactccaac atggacgctg atttatatgg gtataaatgg
     3841 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     3901 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     3961 atggtccgtc tcaactggct gacggagttt atgcctctcc cgaccatcaa gcattttatc
     4021 cgtactcctg atgatgcgtg gttactcacc accgcgattc ctgggaaaac agccttccag
     4081 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc cgtgttcctg
     4141 cgccggttac attcgattcc tgtttgtaat tgtcctttta acagcgatcg tgtatttcgt
     4201 cttgctcagg cgcaatcacg catgaataac ggtttggttg atgcgagtga ttttgatgac
     4261 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcacaagct cttgccattc
     4321 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     4381 gggaaattaa taggttgtat tgatgttgga cgggtcggaa tcgcagaccg ttaccaggac
     4441 cttgccattc tttggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     4501 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     4561 gagtttttct aataatacta gcagaaatca tccttagcga aagctaagga ttttttttat
     4621 ctgattaccg cctttgagtg agcgtcgacc tagtgcggcc gcaagatccg gccacgatgc
     4681 gtccggcgta gaggatctga agatcagcag ttcaacctgt tgatagtacg tactaagctc
     4741 tcatgtttca cgtactaagc tctcatgttt aacgtactaa gctctcatgt ttaacgaact
     4801 aaaccctcat ggctaacgta ctaagctctc atggctaacg tactaagctc tcatgtttca
     4861 cgtactaagc tctcatgttt gaacaataaa attaatataa atcagcaact taaatagcct
     4921 ctaaggtttt aagttttata agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg
     4981 tttaacggtt gtggacaaca agccagggat gtaacgcact gagaagccct tagagcctct
     5041 caaagcaatt ttgagtgaca caggaacact taacggctga catgggaatt aggaagttcc
     5101 tattccgaag ttcctattct ctagaaagta taggaacttc catatgccat gggcccgtgc
     5161 gaatcagaaa taatctcgat atcatctctt ttcttgccga gcgccttcct gagctttgtg
     5221 tcggtgatca ttgaatgggt acacattttt gtttttgggt acacaaaagt gtacacaaag
     5281 ttgcccactc aaagctacac gcaatgtaac actagttcgc agagtgttat ggtttacatc
     5341 cttgaaagcc tgctggataa gggtttagcg taacagaacg tttttacgcg gaattgttcg
     5401 taatatgcca aatgacaatt taagaaagtg ttctaattta ttagaaattt gcacttaaat
     5461 caaaaagtta cggacaattc aaccaccaat caataaatta aagggcacat taaagtacac
     5521 aatatttgtg cccttctctg tttctttccc gttatcagct agctggaaac tttttataca
     5581 gagtttagag ccctactcca tacgtttttg atacgctttg tcgtgattca cctctagctt
     5641 cttcgtctgt ttctactggt attggcacaa acctgattcc aatttgagca aggctatgtg
     5701 ccatctcgat actcgttctt aactcaacag aagatgcttt gtgcatacag cccctcgttt
     5761 attatttatc tcctcagcca gccgctgtgc tttcagtgga tttcggataa cagaaaggcc
     5821 gggaaatacc cagcctcgct ttgtaacgga gtagagacga aagtgattgc gcctacccgg
     5881 atattatcgt gaggatgcgt catcgccatt aattcactga tcagtgatag ctgtcaaaca
     5941 tgagaattga tccggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6001 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6061 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     6121 cgatagcgga gtgtatgctg cacatgacat taacctataa aaataggcgt atcacgaggc
     6181 cctttcgtct tcaagaatta attcccaatt ccaggcatca aataaaacga aaggctcagt
     6241 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     6301 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     6361 gacgcccgcc ataaactgcc aggaattaat tccaggcatc aaataaaacg aaaggctcag
     6421 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     6481 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     6541 ggacgcccgc cataaactgc caggaattaa ttccaggcat caaataaaac gaaaggctca
     6601 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     6661 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     6721 aggacgcccg ccataaactg ccaggaatta attccaggca tcaaataaaa cgaaaggctc
     6781 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     6841 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     6901 caggacgccc gccataaact gccaggaatt ggggatcg
//
