LOCUS       40924_49232        4989 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD050, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  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 4989)
  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 4989)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4989)
  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..4989
                     /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=EcoRV
                     /note="EcoRV"
     misc_feature    408..428
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    429..434
                     /label=XbaI
                     /note="XbaI"
     CDS             complement(600..1634)
                     /codon_start=1
                     /product="repA"
                     /label=repA
                     /note="repA-pDC1; derived from pSUN202; Nostoc sp. PCC8009"
                     /protein_id="AII99670.1"
                     /translation="MANQNRAATQVEGLNHCPLTSQEDRSALSSNVPLPLVLILVKAEI
                     ILQPFMVRLHSGISRQPWARLWYLLRGFDDGSGRVKEIPLVLVEEILGVSQATVYRWLD
                     QGKAVGAFLSGYKVRIGLLTVYLGGLTKVAWHLNLKDWGVVAECPLWEVNANIRALTTG
                     IVTQRLQQRSRYAANRKLKPEYRKSYGAPHPNELLGDEGQSSFKLGAGEVPFVLHVSPS
                     RVFVSKSFIHYGTSQNAVSCKLGIHTRTVRRHQRTLGMEKRQLCQAKYEYAQLDRAVSN
                     EASECWAWSSEGTQSDIGYQSLGISHWVKGRSRFLMASCQEPESSSRTLTSCQRANCLG
                     VSSK"
     misc_feature    1221..1228
                     /label=SwaI
                     /note="SwaI"
     regulatory      complement(1635..2316)
                     /note="PrepA; 682 nts upstream of repA-pDC1; derived from 
                     pSUN202; Nostoc sp. PCC8009"
                     /regulatory_class="promoter"
     misc_feature    2113..2118
                     /label=MfeI
                     /note="MfeI"
     misc_feature    2317..2322
                     /label=SacI
                     /note="SacI"
     protein_bind    2336..2357
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2372..2402
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2410..2426
                     /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             2447..2620
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     misc_feature    2678..2683
                     /label=SacI
                     /note="SacI"
     misc_feature    2684..2704
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons or Escherichia coli origins to be 
                     assembled with a cyanobacterial replicon; synthetic"
     misc_feature    2702..2707
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    2726..2731
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2768..2784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2792..2808)
                     /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(2816..2846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2861..2882)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3170..3758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3932..4789)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4790..4894)
                     /label=AmpR promoter
     misc_feature    4920..4925
                     /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 cgaggatatc gggggccccc
      421 ggggggattc tagaggcttg tgatgattgc tttgcaatcg tctgcctgcg ggacggctcg
      481 attttcgtga aactggcact gtgccagttt acaacgccac ttccgccgct ttgcgcgcat
      541 ggttgttagc ttgaattgct cgcggtaaat gttgcagcga gccatccagg ttttaccctt
      601 catttggaag aaacgcccag acagttcgcc cgctggcagc tggtaagtgt tcggctgctg
      661 ctttctggct cctggcagga tgccatcaga aaacgcgatc gccccttcac ccaatgactg
      721 atacccaatg actgataccc aatgtctgac tgagtgccct cgctactcca agcccagcac
      781 tcagacgctt cattgcttac agcccggtca agttgagcgt attcatactt cgcttggcac
      841 agttgccttt tctccatgcc gagcgttcgc tgatgccgcc gcactgtgcg cgtgtggatg
      901 cccaatttac agctgaccgc attttgagaa gtcccgtaat gaatgaagct tttagaaacg
      961 aacacccgcg acggactgac atgaagaaca aaaggtactt cccccgctcc caatttaaac
     1021 gaggattgcc cctcatcccc caacagttcg tttggatgtg gcgctccgta agacttgcga
     1081 tactccggtt tcagtttgcg gttagccgcg tatctacttc tttgttgcaa acgctgtgtg
     1141 actatgcccg tggtcagcgc tcgaatgttg gcgttgacct cccataacgg acattctgct
     1201 actacccccc aatcttttaa atttaaatgc caagccacct tagtcagtcc gcccaggtaa
     1261 accgtgagca gtccaattct taccttgtat cccgacaaaa atgcccctac tgcttttccc
     1321 tggtcaagcc atctgtacac agtggcttga ctaactccca atatctcctc taccagcacg
     1381 agcggaattt ctttaacccg tccgctccca tcatcaaaac cgcgcaacaa ataccacagc
     1441 cgcgcccaag gttgacggct aattccactg tgtaaccgca ccatgaacgg ctgcaaaatt
     1501 atctcggctt tgactagtat caaaaccaac gggagtggta catttgaact taaggctgaa
     1561 cgatcctcct gtgaagtcag ggggcaatgg ttcagcccct caacttgggt agctgcgcga
     1621 ttttgattgg ccactggttt tcactcaaac ctggttcaat gtgccaatcg cttcactaaa
     1681 tttaagtttt cgcttccgct cgactgaata ctgtgctgat cctcgaaggg tggtaatatg
     1741 tttgaagttc attgcttctg ggtgattaag gaatgattgc ccagtagttt ttccaaatct
     1801 tcaccctatc tctacactgt ttaccgatcc acgcagttgt ataccaaccc caaaacttga
     1861 taaattcaag ttaacttttg ttattgtctg gtttttggat tacggacttg atgggcctgg
     1921 tgaacgactg ctgcatggaa ctctgtaaac atgtgtagac gactgggtaa tcaatttctc
     1981 cactaccgga ctttttctta tataagtttt cttgtgtaag atttgcactt ttagaagtat
     2041 gcgtatccta ttaggaaaac tcacagtttt aagaaaaact gtatttagca tacatcggtt
     2101 agagccgtgt atcaattgac aacatcctat acaggatcta ggttttgcac gtgccataag
     2161 gtgacgaaat atttatttcg tcacataaag cttgaacagt aatcactgca agcagttgaa
     2221 attctacccg tgtagtatgg gtatagaatt ttcctttttc tgtttttttg tagtcaaaat
     2281 ttaggcaaaa catgctactc atttgctact caaaatgagc tcagcgcaac gcaattaatg
     2341 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2401 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctaatga ccatgattac
     2461 ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     2521 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     2581 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc
     2641 ggcaccagaa gcggtgccgg aaagctggct ggagtaggag ctcatccccg ggcccgggcc
     2701 cgatatcctc ggtacccggg gatcctctag agtcgacctg caggcatgca agcttggcgt
     2761 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2821 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2881 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2941 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3001 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3061 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3121 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3361 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3601 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3721 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3781 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4321 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4921 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4981 cctttcgtc
//
