LOCUS       40924_49267        5227 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD057, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5227)
  AUTHORS   Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, 
            Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
  TITLE     Broad-host-range vector system for synthetic biology and 
            biotechnology in cyanobacteria
  JOURNAL   Nucleic Acids Res. (2014) In press
  PUBMED    25074377
REFERENCE   2  (bases 1 to 5227)
  AUTHORS   Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B, 
            Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2014) Division of Biological Sciences, University 
            of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE   3  (bases 1 to 5227)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5227)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. (2014) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-JUN-2014) Division of Biological Sciences, University of 
            California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    405..410
                     /label=ZraI
                     /note="ZraI"
     misc_feature    408..428
                     /note="G5C5; GC-adaptor for seamless assembly, flanking a
                     BHR replicons, neutral sites, or cyanobacterial replicons; 
                     synthetic"
     misc_feature    429..434
                     /label=XbaI
                     /note="XbaI"
     CDS             complement(585..1892)
                     /codon_start=1
                     /product="repA-pFDA"
                     /label=repA-pFDA
                     /note="repA-pFDA; derived from pPL2; 7; Fremyella
                     diplosiphon PCC7601"
                     /protein_id="AII99656.1"
                     /translation="MKHTTFPHSNIIAQSTATEGENFVLQTRLHASILLLTPASKLWYL
                     CRALDLSGRGYVVLSPQNLELLQEKKSTIYRWLKDGKDLGLFRCYSWAGNTLKISLGSL
                     WRACKKARIKNWGTVANNIPLEEILKNNGRRTVATAMTIQDWQERSRYAAKNQLNPLER
                     KCFEIPATADVLTPQTSPKLTSGGTTGVLHVGEKRIFVGRSFIPFGVSQKRISNELNSQ
                     PASCGVSVRTVQRHVERLEVKHRQLMQARREYREIAHRIKQGATSWQCKSDADISFTWG
                     NQPDEIVLHERNGKSSARREGGHTIKLDQLCNYSNTHWRYRCNLYLLSYELTSMRTTRY
                     QWKKRGLNVKIAPVENCPSEISPQTPEMLESPKLHAEGRGLGGQNKSVKNENPQEVDTV
                     SNAFAPGGSEWAKTKAMLLAKQAERRQKRWDSLNSI"
     misc_feature    1152..1157
                     /label=MfeI
                     /note="MfeI"
     regulatory      complement(1893..2554)
                     /note="PrepA; 662 nts upstream of repA-pFDA; derived from 
                     pPL2.7; Fremyella diplosiphon PCC7601"
                     /regulatory_class="promoter"
     misc_feature    2417..2422
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    2555..2560
                     /label=SacI
                     /note="SacI"
     protein_bind    2574..2595
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2610..2640
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2648..2664
                     /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)."
     CDS             2685..2858
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     misc_feature    2916..2921
                     /label=SacI
                     /note="SacI"
     misc_feature    2922..2942
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons; synthetic"
     misc_feature    2940..2945
                     /label=ZraI
                     /note="ZraI"
     misc_feature    2964..2969
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(3006..3022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3030..3046)
                     /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(3054..3084)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3099..3120)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3408..3996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4170..5027)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5028..5132)
                     /label=AmpR promoter
     misc_feature    5158..5163
                     /label=ZraI
                     /note="ZraI"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgaggacgtc gggggccccc
      421 ggggggattc tagattccac tgctactagt ggcgctagtg gcgctcacag gattaaataa
      481 gggcataaaa atacctttgt tttaagtaaa aatcacttcg cccagcataa tctcatcgct
      541 tcaaaaaagc gatcgcctgt gaataaatag ctgaaatgct tacatcatat agagttcaga
      601 ctatcccatc gtttttgcct tctctcggct tgcttggcca gtagcatagc cttagttttt
      661 gcccactcag aaccccctgg agcgaacgca ttgcttactg tgtctacttc ctggggattc
      721 tcatttttga cactcttatt ctgacccccc aaaccacgcc cttcggcgtg aagtttgggg
      781 gattcaagca tttcgggggt ttggggggaa atctcacttg ggcagttttc cacaggtgca
      841 attttgacgt ttaacccgcg tttcttccac tgatagcgag ttgtcctcat cgaggtcaac
      901 tcgtagctga gaaggtacaa attacagcga tagcgccaat gtgtattgct gtagttgcac
      961 agctgatcca gtttgattgt gtgccccccc tcacgcctag ctgatgattt cccgtttctc
     1021 tcatggagta caatttcatc gggctgattg ccccaggtga agctgatgtc agcatcactc
     1081 ttgcactgcc agctagtagc cccctgcttg attctgtgag cgatttcccg gtattcccgg
     1141 cgggcttgca tcaattgcct gtgttttacc tccaaacgct caacgtgtct ttggactgtg
     1201 cgaactgaaa caccacagct ggccggctgg gagtttagct cattgctgat ccgcttctga
     1261 gaaaccccaa aggggatgaa cgagcggcca acaaaaatcc gcttctcgcc aacgtgtaac
     1321 actccagttg ttcccccact agtcagcttc ggtgaggttt ggggtgttag cacgtcggca
     1381 gtagccggaa tctcaaaaca tttacgctca agcgggttaa gttgattttt tgccgcgtat
     1441 cgtgagcgct cttgccaatc ttgaattgtc atcgctgtag cgacagtgcg acgaccatta
     1501 ttttttaaaa tttcttctag tgggatgttg ttggccactg tgccccaatt ttttattctt
     1561 gcttttttgc aagcgcgcca caggctaccc aggctaattt ttaaagtgtt ccccgcccag
     1621 gagtaacaac gaaatagccc caaatcctta ccatccttga gccaacggta aattgttgat
     1681 tttttttcct gcaagagttc cagattttga ggagacaaaa ctacataacc acgaccactc
     1741 agatccaacg ctcgacacaa ataccacaat ttagaagccg gagtgagcag gagaattgaa
     1801 gcgtgtaatc ttgtttgtag tacaaaattc tccccctctg tagcggtgga ttgtgctata
     1861 atattggaat gtggaaatgt agtatgtttc acaaatcgcc ccttttatag cgacttgctg
     1921 acacatcacc ccaagctttc caacagcttt atattttcac gctcaatatt tgttaatttt
     1981 tagttagaga tttatttttg ttggcttgtt ggcgcgtctg ggttttttga tgtgtctgca
     2041 agtcattttt ttatgtgtcg cttcatgtca ggttgtcatg gtgttgtagc gtttattgca
     2101 tttattgctt gcaatttcct cctaattgaa ttaaggaatt aatggggcga tttaaaaatc
     2161 gccctggtgt ctacattcca gtagtttgcg gctactggga aataccgctc tctagcggca
     2221 cttcaaattt atgataagcg aaaatgaagg gttttcgctt gatagtaccg tgtcggcttt
     2281 tttacaaatt cggaaagccc ttggactgtc acgtagccag tttgggctgg aactgggtat
     2341 tgatgccgat acaataggga aatgggagaa ttcaaaagtt aacgtagcgc taactatcgc
     2401 ccaaatcaag cgattagata tctatttaaa agaattaggg ttatcctggc aggacttgcc
     2461 agatcatgtt ggcaacccag caatcgcccc gccagcatac aagcgcatca gaaaaaaaat
     2521 taaataacca tgtatatacg tggctagtca ttttgagctc agcgcaacgc aattaatgtg
     2581 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     2641 tgtggaattg tgagcggata acaatttcac acaggaaaca gctaatgacc atgattacgg
     2701 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2761 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     2821 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgctttgcc tggtttccgg
     2881 caccagaagc ggtgccggaa agctggctgg agtaggagct catccccggg cccgggcccg
     2941 acgtcctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa
     3001 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3061 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3121 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3181 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3241 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3301 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3361 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3421 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3481 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3541 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3601 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3841 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4021 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4081 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4141 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4201 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4261 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4321 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4381 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4441 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4501 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4561 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4621 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4681 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4741 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4801 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4861 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4921 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4981 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5101 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5161 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     5221 tttcgtc
//
