LOCUS       40924_33882        6398 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Chromosomal integration vector pOSIP-CH, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6398)
  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 6398)
  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 6398)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6398)
  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..6398
                     /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)"
     CDS             2270..3340
                     /label=HK022 integrase
                     /note="integrase from phage HK022"
     promoter        3442..3544
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             3545..4201
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      4225..4319
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4376..4764
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     misc_feature    4775..4822
                     /note="FRT; Flp recombinase target site"
     protein_bind    4775..4822
                     /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(4852..5078)
                     /label=HK022 attP
                     /note="attachment site of phage HK022"
     terminator      complement(5113..5359)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     terminator      5679..5765
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5860..5946
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6041..6127
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(6225..6268)
                     /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 cgcgtgtagc cttttgaaga ggatcagaaa tgggaagaag
     2281 gcgaagtcat gagcgccggg atttaccccc taacctttat ataagaaaca atggatatta
     2341 ctgctacagg gacccaagga cgggtaaaga gtttggatta ggccgagaca ggcgaatcgc
     2401 aatcactgaa gctatacagg ccaacattga gttgctatcc gggaacaggc gtgagtcact
     2461 gatagacaga attaaaggcg ctgacgcaat cactcttcat gcgtggcttg accgatatga
     2521 aacaatcctc agcgagaggg gtatcaggcc gaaaactcta ctcgactacg ccagcaaaat
     2581 cagagcaatc cgaagaaaat tgccggacaa accgctcgct gacatatcaa cgaaagaggt
     2641 ggcagcaatg ctaaacacct acgtagcaga aggcaaatcg gcttccgcaa aattgatcag
     2701 gtcaaccctt gttgacgttt ttcgtgaggc aatagccgag gggcatgtgg ctacgaatcc
     2761 ggtaacagca acccgcacag caaaatcaga agtaaggcgt tcaagactga cagctaatga
     2821 atatgtcgcg atataccatg cagccgaacc gctcccaatc tggctgagac tggcaatgga
     2881 tttggctgtc gttacagggc agagagtagg cgatttgtgc agaatgaaat ggtcagacat
     2941 aaacgacaac catcttcaca ttgaacagag taaaacaggg gctaaactcg ccattccact
     3001 aacgctaacg attgacgcgc tcaatatctc attggctgat acactacagc aatgcaggga
     3061 ggccagcagc agtgaaacta taatcgcatc aaagcatcac gatccgcttt ccccgaaaac
     3121 agtatcaaag tattttacaa aggcgagaaa tgcatctgga ctctcatttg atggaaaccc
     3181 gccaacattc catgaactgc gtagcctgtc agcgaggcta taccggaacc agattggcga
     3241 taagtttgct caacgtcttc tcgggcataa atcagattca atggcggcgc ggtataggga
     3301 cagccgtgga cgggaatggg acaaaattga aatcgacaaa tgattttatt ttgactaata
     3361 atgacctact taccatgggt atggacagtt ttccctttga tatgtaacgc acgttgtgga
     3421 tatctggcga aaatgagacg ttgatcggca cgtaagaggt tccaactttc accataatga
     3481 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     3541 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     3601 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     3661 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     3721 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     3781 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     3841 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     3901 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     3961 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     4021 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     4081 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgtct gtgatggctt
     4141 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     4201 gtaatttgat atcgagctcg cttggactcc tgttgataga tccagtaatg acctcagaac
     4261 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
     4321 caagcgtcga cctagtgcgg ccgcaagatc cggccacgat gcgtccggcg tagaggatct
     4381 gaagatcagc agttcaacct gttgatagta cgtactaagc tctcatgttt cacgtactaa
     4441 gctctcatgt ttaacgtact aagctctcat gtttaacgaa ctaaaccctc atggctaacg
     4501 tactaagctc tcatggctaa cgtactaagc tctcatgttt cacgtactaa gctctcatgt
     4561 ttgaacaata aaattaatat aaatcagcaa cttaaatagc ctctaaggtt ttaagtttta
     4621 taagaaaaaa aagaatatat aaggctttta aagcttttaa ggtttaacgg ttgtggacaa
     4681 caagccaggg atgtaacgca ctgagaagcc cttagagcct ctcaaagcaa ttttgagtga
     4741 cacaggaaca cttaacggct gacatgggaa ttaggaagtt cctattccga agttcctatt
     4801 ctctagaaag tataggaact tccatatgcc atgggacaaa attgaaatcg acaaatgatt
     4861 ttattttgac taataatgac ctacttacat taatttactg ataattaaag agattttaaa
     4921 tatacaactt attcacctaa aggatgacaa aataacatta atcacttaaa aatcatcgca
     4981 ttacactaat ctgtggttaa atgatagact acataatgcg acaaaacgca acatatccag
     5041 tcactatgaa tcaactactt agatagtatt agtgacctga gacagagcat tagctagctt
     5101 cttcgtctgt ttctactggt attggcacaa acctgattcc aatttgagca aggctatgtg
     5161 ccatctcgat actcgttctt aactcaacag aagatgcttt gtgcatacag cccctcgttt
     5221 attatttatc tcctcagcca gccgctgtgc tttcagtgga tttcggataa cagaaaggcc
     5281 gggaaatacc cagcctcgct ttgtaacgga gtagagacga aagtgattgc gcctacccgg
     5341 atattatcgt gaggatgcgt catcgccatt aattcactga tcagtgatag ctgtcaaaca
     5401 tgagaattga tccggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5461 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5521 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     5581 cgatagcgga gtgtatgctg cacatgacat taacctataa aaataggcgt atcacgaggc
     5641 cctttcgtct tcaagaatta attcccaatt ccaggcatca aataaaacga aaggctcagt
     5701 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     5761 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     5821 gacgcccgcc ataaactgcc aggaattaat tccaggcatc aaataaaacg aaaggctcag
     5881 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     5941 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     6001 ggacgcccgc cataaactgc caggaattaa ttccaggcat caaataaaac gaaaggctca
     6061 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     6121 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     6181 aggacgcccg ccataaactg ccaggaatta attccaggca tcaaataaaa cgaaaggctc
     6241 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     6301 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     6361 caggacgccc gccataaact gccaggaatt ggggatcg
//
