LOCUS       40924_33887        6392 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Chromosomal integration vector pOSIP-CO, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6392)
  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 6392)
  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 6392)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6392)
  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..6392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             142..444
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     rep_origin      477..1065
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1125..1145
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1200..1227
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1276..1307)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     protein_bind    1384..1417
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(1427..2137)
                     /label=lambda repressor (ts)
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     RBS             2226..2234
                     /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)"
     RBS             2246..2268
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2276..3283
                     /label=phage 186 integrase
                     /note="integrase from phage 186"
     promoter        3387..3489
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             3490..4146
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      4170..4264
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4321..4709
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     misc_feature    4720..4767
                     /note="FRT; Flp recombinase target site"
     protein_bind    4720..4767
                     /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)."
     protein_bind    complement(4780..4994)
                     /label=phage 186 attP
                     /note="attachment site of phage 186"
     terminator      complement(5107..5353)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     terminator      5673..5759
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5854..5940
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6035..6121
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(6219..6262)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gaattcgagc tcggtacccg gggatcctct agagggctta ctaaaagcca gataacagta
       61 tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag
      121 tatgtcaaaa agaggtatgc tatgcagttt aaggtttaca cctataaaag agagagccgt
      181 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
      241 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
      301 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
      361 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
      421 attaacctga tgttctgggg aatataataa cccgtagaaa agatcaaaag atcttcttga
      481 gatccttttt ttctgcgcgt aatctgctac ttgcaaacaa aaaaaccacc gctaccagcg
      541 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggctttagc
      601 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
      661 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
      721 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
      781 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
      841 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
      901 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
      961 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1021 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1081 gcctttttac ggttcctggc cttttgctgg ccttttgctc acattactag tagcggccgc
     1141 tgcaggcatg cctcgagatg catggcgcct aacctaaact gacaggcatc aaattaagca
     1201 gaaggccatc ctgacggatg gcctttttgc gtttcgaaca attgaaaaaa cctcgcgcct
     1261 tacctgttga gtaatagtca aaagcctccg gtcggaggct tttgactttc tgcttactga
     1321 atttcggtgg tgccgttaat taaccggtgg gccctcatga taataatggt ttcttagacg
     1381 tccgaagttc ctattctcta gaaagtatag gaacttccct aggtcagcca aacgtctctt
     1441 caggccactg actagcgata actttcccca caacggaaca actctcattg catgggatca
     1501 ttgggtactg tgggtttagt ggttgtaaaa acacctgacc gctatccctg atcagtttct
     1561 tgaaggtaaa ctcatcaccc ccaagtctgg ctatgcagaa atcacctggc tcaacagcct
     1621 gctcagggtc aacgagaatt aacattccgt caggaaagct tggcttggag cctgttggtg
     1681 cggtcatgga attaccttca acctcaagcc agaatgcaga atcactggct tttttggttg
     1741 tgcttaccca tctctccgca tcacctttgg taaaggttct aagcttaggt gagaacatcc
     1801 ctgcctgaac atgagaaaaa acagggtact catactcact tctaagtgac ggctgcatac
     1861 taaccgcttc atacatctcg tagatttctc tggcgattga agggctaaat tcttcaacgc
     1921 taactttgag aatttttgta agcaatgcgg cgttataagc atttaatgca ttgatgccat
     1981 taaataaagc accaacgcct gactgcccca tccccatctt gtctgcgaca gattcctggg
     2041 ataagccaag ttcatttttc tttttttcat aaattgcttt aaggcgacgt gcgtcctcaa
     2101 gctgctcttg tgttaatggt ttcttttttg tgctcatacg ttaaatctat caccgcaagg
     2161 gataaatatc taacaccgtg cgtgttgact attttacctc tggcggtgat aatggttgca
     2221 tgtactaagg aggttgtata cgcgttttgt ttaactttaa gaaggagata tacatatgac
     2281 cgtccgtaaa aatccggctg gcggttggat ttgtgaactc tacccaaacg gtgcaaaagg
     2341 caaacgtatc agaaagaaat tcgctactaa aggcgaggct ctggcgtttg agcagtacac
     2401 cgttcaaaac ccgtggcagg aagaaaagga agacaggcgc acgttaaaag agctggttga
     2461 ttcatggtat agcgctcatg gcattacact gaaagatggt ttgaaacgcc agttagccat
     2521 gcaccatgct tttgagtgta tgggcgaacc actcgcacgc gatttcgatg cgcagatgtt
     2581 ttcccgctac cgagaaaaac ggttaaaagg tgagtatgcc cgttcaaaca gagtgaaaga
     2641 ggtatcgcct cgcacgctta atcttgagct ggcctacttc cgggcggtgt tcaatgagct
     2701 aaaccgcctc ggagaatgga agggtgaaaa cccactaaaa aatatgcgcc cattccgcac
     2761 agaagaaatg gaaatgactt ggctaactca cgaccaaatt tcgcaactgc tcggagagtg
     2821 taaccgacat gaccaccctg atttagaaac cgtggtaaga atctgtctcg caactggcgc
     2881 acggtggtct gaggccgaga gtctgagaaa aagccaactc gcgaaataca aaatcacata
     2941 caccaacacg aaaggtagaa aaaatcgcac cgtcccaatc agcaaagagc tctatgagtc
     3001 tctgcctgat gataaaaagg gccggttgtt cagtgattgt tatggcgcgt tccggtcagc
     3061 tttggaaaga acaggtatcg aactaccggc aggacaactt acccacgttt tacgtcacac
     3121 cttcgctagt cactttatga tgaatggcgg taatattttg gtgttgcagc gcgtactcgg
     3181 ccatacagat ataaaaatga ccatgcgata tgcgcacttt gcacccgacc atttagagga
     3241 tgcagttaaa ctaaatcctt tagttcacat tactaacagc aaataagtta tggaggttgt
     3301 atgtcagttc ggctagccca tgggtatgga cagttttccc tttgatatgt aacgcacgtt
     3361 gtggatatct ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa ctttcaccat
     3421 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     3481 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     3541 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     3601 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     3661 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg tatggcaatg
     3721 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     3781 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     3841 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     3901 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     3961 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     4021 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtctgtgat
     4081 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     4141 ggggcgtaat ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc
     4201 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
     4261 gaatccaagc gtcgacctag tgcggccgca agatccggcc acgatgcgtc cggcgtagag
     4321 gatctgaaga tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt
     4381 actaagctct catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc
     4441 taacgtacta agctctcatg gctaacgtac taagctctca tgtttcacgt actaagctct
     4501 catgtttgaa caataaaatt aatataaatc agcaacttaa atagcctcta aggttttaag
     4561 ttttataaga aaaaaaagaa tatataaggc ttttaaagct tttaaggttt aacggttgtg
     4621 gacaacaagc cagggatgta acgcactgag aagcccttag agcctctcaa agcaattttg
     4681 agtgacacag gaacacttaa cggctgacat gggaattagg aagttcctat tccgaagttc
     4741 ctattctcta gaaagtatag gaacttccat atgccatggg cgatggttct gagtaacaga
     4801 taatagaatg gcgataaagt ggcggtagaa atggcgaata ataggtaatt attggcaaac
     4861 aatggcaatc tatgtcaatg ataaataacg catactattg attttcggtt gttcttgtag
     4921 gaactcataa tcgcttggtc gctggttcaa gtccagcagg ggccaccaaa tttaagtagt
     4981 aaatacatac agttaggcca ctctctcgag tggccttttt gttgcctcgt tttcgagtgg
     5041 cgatagaatg gcggtggatt ttttaccgcc actttcctga gtgcatgcta gcttcttcgt
     5101 ctgtttctac tggtattggc acaaacctga ttccaatttg agcaaggcta tgtgccatct
     5161 cgatactcgt tcttaactca acagaagatg ctttgtgcat acagcccctc gtttattatt
     5221 tatctcctca gccagccgct gtgctttcag tggatttcgg ataacagaaa ggccgggaaa
     5281 tacccagcct cgctttgtaa cggagtagag acgaaagtga ttgcgcctac ccggatatta
     5341 tcgtgaggat gcgtcatcgc cattaattca ctgatcagtg atagctgtca aacatgagaa
     5401 ttgatccggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5461 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5521 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     5581 cggagtgtat gctgcacatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5641 gtcttcaaga attaattccc aattccaggc atcaaataaa acgaaaggct cagtcgaaag
     5701 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     5761 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
     5821 cgccataaac tgccaggaat taattccagg catcaaataa aacgaaaggc tcagtcgaaa
     5881 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     5941 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     6001 ccgccataaa ctgccaggaa ttaattccag gcatcaaata aaacgaaagg ctcagtcgaa
     6061 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     6121 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     6181 cccgccataa actgccagga attaattcca ggcatcaaat aaaacgaaag gctcagtcga
     6241 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     6301 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     6361 gcccgccata aactgccagg aattggggat cg
//
