LOCUS       40924_5929        4494 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Integrative vector pBaSysBioII, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4494)
  AUTHORS   Botella E, Fogg M, Jules M, Piersma S, Doherty G, Hansen A, Denham 
            EL, Le Chat L, Veiga P, Bailey K, Lewis PJ, van Dijl JM, Aymerich S,
            Wilkinson AJ, Devine KM.
  TITLE     pBaSysBioII: an integrative plasmid generating gfp transcriptional 
            fusions for high-throughput analysis of gene expression in Bacillus 
            subtilis
  JOURNAL   Microbiology (Reading, Engl.) 156 (PT 6), 1600-1608 (2010)
  PUBMED    20150235
REFERENCE   2  (bases 1 to 4494)
  AUTHORS   Botella E.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-OCT-2009) Smurfit Institute of Genetics, Trinity 
            College Dublin, Lincoln Place, Dublin 2, Ireland
REFERENCE   3  (bases 1 to 4494)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4494)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microbiology (Reading, Engl.) 156 (PT 6), 1600-1608 (2010)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-OCT-2009) Smurfit Institute of Genetics, Trinity College Dublin,
            Lincoln Place, Dublin 2, Ireland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     145..161
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    184..212
                     /label=LIC site
                     /note="LIC site"
     CDS             216..929
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      952..1045
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     gene            1082..2233
                     /gene="aad9"
                     /label=aad9
     CDS             1352..2119
                     /codon_start=1
                     /gene="aad9"
                     /product="spectinomycin adenyltransferase"
                     /label=aad9
                     /note="confers resistance to spectinomycin"
                     /protein_id="ACY68260.1"
                     /translation="MRRIYLNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNS
                     DLDFLVVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQ
                     EFIYGEWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVR
                     RAIMDSSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERI
                     LLAVRSYLGENIEWTNENVNLTINYLNNRLKKL"
     primer_bind     complement(2276..2292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2300..2316)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2324..2354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2369..2390)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2678..3266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3440..4297)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4298..4402)
                     /label=AmpR promoter
ORIGIN
        1 agcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
       61 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      121 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgcca cgcgtcgact
      181 tttaccgcgg gctttcccgg gaaggaggaa ctactatgcg taaaggagaa gaacttttca
      241 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
      301 tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca
      361 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtgttc
      421 aatgctttgc gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc
      481 ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac
      541 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
      601 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aactcacaca
      661 atgtatacat catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac
      721 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
      781 gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga
      841 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
      901 ttacacatgg catggatgaa ctatacaaat aagcttaatt agctgagctt ggactcctgt
      961 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     1021 gccgggcgtt ttttattggt gagaagtcga ctacagaacc gaattcctgc agctggcgaa
     1081 tggcgatttt cgttcgtgaa tacatgttat aataactata actaataacg taacgtgact
     1141 ggcaagagat atttttaaaa caatgaatag gtttacactt actttagttt tatggaaatg
     1201 aaagatcata tcatatataa tctagaataa aattaactaa aataattatt atctagataa
     1261 aaaatttaga agccaatgaa atctataaat aaactaaatt aagtttattt aattaacaac
     1321 tatggatata aaataggtac taatcaaaat agtgaggagg atatatttga atacatacga
     1381 acaaattaat aaagtgaaaa aaatacttcg gaaacattta aaaaataacc ttattggtac
     1441 ttacatgttt ggatcaggag ttgagagtgg actaaaacca aatagtgatc ttgacttttt
     1501 agtcgtcgta tctgaaccat tgacagatca aagtaaagaa atacttatac aaaaaattag
     1561 acctatttca aaaaaaatag gagataaaag caacttacga tatattgaat taacaattat
     1621 tattcagcaa gaaatggtac cgtggaatca tcctcccaaa caagaattta tttatggaga
     1681 atggttacaa gagctttatg aacaaggata cattcctcag aaggaattaa attcagattt
     1741 aaccataatg ctttaccaag caaaacgaaa aaataaaaga atatacggaa attatgactt
     1801 agaggaatta ctacctgata ttccattttc tgatgtgaga agagccatta tggattcgtc
     1861 agaggaatta atagataatt atcaggatga tgaaaccaac tctatattaa ctttatgccg
     1921 tatgatttta actatggaca cgggtaaaat cataccaaaa gatattgcgg gaaatgcagt
     1981 ggctgaatct tctccattag aacataggga gagaattttg ttagcagttc gtagttatct
     2041 tggagagaat attgaatgga ctaatgaaaa tgtaaattta actataaact atttaaataa
     2101 cagattaaaa aaattataaa aaaattgaaa aaatggtgga aacacttttt tcaatttttt
     2161 tgttttatta tttaatattt gggaaatatt cattctaatt ggtaatcaga ttttagaaaa
     2221 caataaaccc ttgcataggg ggatctcgag gcctcgcgag cttggcgtaa tcatggtcat
     2281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2341 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2461 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2521 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2581 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2641 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2701 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2761 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2821 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2881 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2941 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3001 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3061 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3121 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3181 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3241 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3301 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3361 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3421 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3481 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3541 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3601 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3661 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3721 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3781 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3841 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3901 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3961 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4021 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4081 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4141 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4201 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4261 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4321 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4381 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac tgccgggcct
     4441 cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgct
//
