LOCUS       40924_49327        6207 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD076, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  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 6207)
  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 6207)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6207)
  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..6207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..6
                     /label=ZraI
                     /note="ZraI"
     misc_feature    4..24
                     /note="G5C5; GC-adaptor for seamless assembly, flanking a
                     BHR replicons, neutral sites, or cyanobacterial replicons; 
                     synthetic"
     misc_feature    25..30
                     /label=XbaI
                     /note="XbaI"
     misc_recomb     complement(31..1035)
                     /note="A7120NS2-LA; left arm of the homologous
                     recombination site NS2 (all1997_D43-all1697_D1) into the 
                     chromosome of Anabaena sp. PCC7120 derived from INSD 
                     accession NC_003272"
     rep_origin      1072..1660
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1846..1986)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    2149..2154
                     /label=EcoRV
                     /note="EcoRV"
     CDS             complement(2177..3595)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     regulatory      complement(3596..3901)
                     /note="PsacB; derived from Bacillus subtilis INSD accession
                     AAA72307"
                     /regulatory_class="promoter"
     misc_recomb     complement(3902..4951)
                     /note="A7120NS2-RA; right arm of the homologous
                     recombination site NS2 (all1997_D43-all1697_D1) into the 
                     chromosome of Anabaena sp. PCC7120 derived from INSD 
                     accession NC_003272"
     misc_feature    4952..4957
                     /label=NheI
                     /note="NheI"
     misc_feature    4958..4978
                     /note="GC; GC-adaptor for seamless assembly, flanking BHR 
                     replicons or neutral sites; synthetic"
     misc_feature    4976..4981
                     /label=ZraI
                     /note="ZraI"
     CDS             complement(5105..5962)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5963..6067)
                     /label=AmpR promoter
     misc_feature    6093..6098
                     /label=ZraI
                     /note="ZraI"
ORIGIN
        1 gacgtcgggg gcccccgggg ggattctaga gctttgctta ccctagtgag atgcaaacaa
       61 caataattta ggacttaccc atgaaaacga aaaatcaaga gtttagacaa gggtgtagga
      121 gtataaattt ttaagagcct tacacccagt ctcaacagac aacctgggtg agtaagacct
      181 gttcatatca tatttagtct gaacctgatc cagacttggt tagatttatc aattcttaat
      241 ttaatttttt gagtatttta actgattgac aaaatttact gcgcttttaa ttctcaccaa
      301 cgctcttgga atacactatt tttgtggaga ttacaaaatt ttttggtgtc aatgacattc
      361 ctgatttatg ttgttttgat taaaaaattc tttaaagaga caaaatttta gaaattactg
      421 agatgatctc ctaaagatat ttgttcccta tatagatttt aatttttgct tgactcgctt
      481 aggttcgcta aatccaattt tgcggagcct tttgatgtct ttaactgatg acctgctgca
      541 agctttaaag ttaacattac cggacaacaa cgtaaacatc gtagttgctt atcctggttc
      601 tggtggtgat ggaccaagct acgtagacct tacaatctcc aacggtaatt tcctcaatgg
      661 actgtatgac acctattgca ttgatgcgaa tcaaactatt gttccagggc aatcctacca
      721 agcgcaagtt tattctagct ataatgaccc tctgccaaca gggataattg ataaacccga
      781 aaacctagat gagattaact ggttaattaa ccagaaattc gtaggcaaac aatcttcagg
      841 gggtctcggt gcctatacct ccggtgatat acaggtagcc atctggactc tgcttgagaa
      901 cacagttcct tccaacatat taggtacagt aggcgcatac aaccaaaacc gagccaatca
      961 gattgttgaa gcagcctttc aaaatggtgg aggctttgta cctggtatca gccaagatgg
     1021 catacctcaa gatatagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1081 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1141 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1201 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1261 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1321 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1381 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1441 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1501 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1561 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1621 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1681 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1741 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1801 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     1861 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     1921 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     1981 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2041 gcatccgctt acagacaagc tccgcccggt tagccatttg cctgctttta tatagttcat
     2101 atgggattca cctttatgtt gataagaaat aaaagaaaat gccaatagga tatcggcatt
     2161 ttcttttgcg tttttatttg ttaactgtta attgtccttg ttcaaggatg ctgtctttga
     2221 caacagatgt tttcttgcct ttgatgttca gcaggaagct tggcgcaaac gttgattgtt
     2281 tgtctgcgta gaatcctctg tttgtcatat agcttgtaat cacgacattg tttcctttcg
     2341 cttgaggtac agcgaagtgt gagtaagtaa aggttacatc gttaggatca agatccattt
     2401 ttaacacaag gccagttttg ttcagcggct tgtatgggcc agttaaagaa ttagaaacat
     2461 aaccaagcat gtaaatatcg ttagacgtaa tgccgtcaat cgtcattttt gatccgcggg
     2521 agtcagtgaa caggtaccat ttgccgttca ttttaaagac gttcgcgcgt tcaatttcat
     2581 ctgttactgt gttagatgca atcagcggtt tcatcacttt tttcagtgtg taatcatcgt
     2641 ttagctcaat cataccgaga gcgccgtttg ctaactcagc cgtgcgtttt ttatcgcttt
     2701 gcagaagttt ttgactttct tgacggaaga atgatgtgct tttgccatag tatgctttgt
     2761 taaataaaga ttcttcgcct tggtagccat cttcagttcc agtgtttgct tcaaatacta
     2821 agtatttgtg gcctttatct tctacgtagt gaggatctct cagcgtatgg ttgtcgcctg
     2881 agctgtagtt gccttcatcg atgaactgct gtacattttg atacgttttt ccgtcaccgt
     2941 caaagattga tttataatcc tctacaccgt tgatgttcaa agagctgtct gatgctgata
     3001 cgttaacttg tgcagttgtc agtgtttgtt tgccgtaatg tttaccggag aaatcagtgt
     3061 agaataaacg gatttttccg tcagatgtaa atgtggctga acctgaccat tcttgtgttt
     3121 ggtcttttag gatagaatca tttgcatcga atttgtcgct gtctttaaag acgcggccag
     3181 cgtttttcca gctgtcaata gaagtttcgc cgactttttg atagaacatg taaatcgatg
     3241 tgtcatccgc atttttagga tctccggcta atgcaaagac gatgtggtag ccgtgatagt
     3301 ttgcgacagt gccgtcagcg ttttgtaatg gccagctgtc ccaaacgtcc aggccttttg
     3361 cagaagagat atttttaatt gtggacgaat cgaattcagg aacttgatat ttttcatttt
     3421 tttgctgttc agggatttgc agcatatcat ggcgtgtaat atgggaaatg ccgtatgttt
     3481 ccttatatgg cttttggttc gtttctttcg caaacgcttg agttgcgcct cctgccagca
     3541 gtgcggtagt aaaggttaat actgttgctt gttttgcaaa ctttttgatg ttcatcgttc
     3601 atgtctcctt ttttatgtac tgtgttagcg gtctgcttct tccagccctc ctgtttgaag
     3661 atggcaagtt agttacgcac aataaaaaaa gacctaaaat atgtaagggg tgacgccaaa
     3721 gtatacactt tgccctttac acattttagg tcttgcctgc tttatcagta acaaacccgc
     3781 gcgatttact tttcgacctc attctattag actctcgttt ggattgcaac tggtctattt
     3841 tcctcttttg tttgatagaa aatcataaaa ggatttgcag actacgggcc taaagaacta
     3901 gcaactgtat ctcctgcacc tgctccgcct gcacctcccg ctcccactcc tgaaagcacc
     3961 ccgaagcaat aaagaaaact ctaaccccca atttttaggg ggttattgat gagtaggata
     4021 agaattttaa atgttgaatt gttacactcc ttcaatagcg gcgacaatac caaaaatcaa
     4081 aaataggcat ccgccggcaa aggtaagctg acgttcagaa attttcccgg caatcatttt
     4141 gcccccaatg acagcgatcg ccgcacaaat ggcatgacct aaaatggctc ctatggtgac
     4201 accaataatg ttattacctg cggctaaggc tatggtggct atttgggtgc gatcgcccca
     4261 ctctgccata aaggtcaaga caaaggcttc taatataatt gataagggcg ttttttgttt
     4321 tggtagctgt aaatctgctt tttctacggc tgctttcgcc tcttccatct cttccatcac
     4381 ttcagcttta tccgccgccg ccgacatctt actagcgtca tacaacagtt tcagaccaaa
     4441 ggcaataaat aaggttattt cagcgtaata aatgtacact ggtggcaata gagatgcagc
     4501 tttgccaaat aataccgaga gtattgtcat tgcggctaaa gcgcccacca ctcctgtaaa
     4561 tacccatcgt cgggaatggt gcattgccaa gatcatggcg atgaagaaag ttttatctcc
     4621 cagttctgaa actgtaatta gtaataaacc tgcggtaaaa gctgttagca ctctctcaag
     4681 ctcctgaaaa atgtttaccg atgtgaagct tgatgaatgc caaaagaccc caccaagcca
     4741 catcttaaat gtgaacttag tgaaggtctc gcttccaaat attttctagt tgtcacctga
     4801 accaggctca tcgccagtat gttgattcag atgtactggc ttttgccagc agctactccc
     4861 cttcttatca gaataaaatt gtatagcaat tctcaatgaa ttgtaaatat atttttagct
     4921 gttgagtgca aatatcagga catatccacc agctagcatc gcgcgcgcgc gcgcggacgt
     4981 cctcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5041 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5101 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5161 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5221 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5281 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5341 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5401 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5461 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5521 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5581 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5641 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5701 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5761 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5821 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5881 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5941 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6001 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6061 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6121 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     6181 acggtgaaaa cctctgacac atgcagc
//
