LOCUS       40924_49277        4989 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD059, 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    complement(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"
     terminator      complement(442..469)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    478..502
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(522..1238)
                     /label=mEGFP
                     /note="enhanced GFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
     CDS             complement(1285..1587)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     regulatory      complement(1843..1892)
                     /note="PccdB; upstream region of the ccdB ORF including
                     promoter and RBS; derived from the destination cassette 
                     used in the Gateway Technology developed by Life 
                     Technologies"
                     /regulatory_class="promoter"
     CDS             complement(1932..2588)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(2642..2672)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     misc_feature    complement(2684..2704)
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; 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 cccccggggg
      421 cccccgattc tagaagcttc aaaaaggcca tccgtcagga tggccttctc ctgcaggacc
      481 actttgtaca agaaagctgg gtcggcgcgc ccaccctttt atttgtacaa ttcatccata
      541 ccatgggtaa taccagcagc agtaacaaat tctaacaaga ccatgtggtc tctcttttcg
      601 tttggatctt tggataattt agattgagtg gataagtaat ggttgtctgg taacaagact
      661 ggaccatcac caattggagt attttgttga taatggtcag ctaattgaac agaaccatct
      721 tcaatgttgt gtctaatttt gaagttaact ttgataccat tcttttgttt gtcagccatg
      781 atgtaaacat tgtgagagtt atagttgtat tccaatttgt gacctaaaat gttaccatct
      841 tctttaaaat caataccttt taattcgatt ctattaacta aggtatcacc ttcaaacttg
      901 acttcagctc tggtcttgta gttaccgtca tctttgaaaa aaatagttct ttcttgaaca
      961 taaccttctg gcatggcaga cttgaaaaag tcatgttgtt tcatatgatc tgggtatcta
     1021 gaaaaacatt gaacaccata agttaaagta gtgactaagg ttggccatgg aactggcaat
     1081 ttaccagtag tacaaataaa ttttaaggtc aatttaccgt aagtagcatc accttcacct
     1141 tcaccggaga cagaaaattt gtgaccatta acatcaccat ctaattcaac caaaattggg
     1201 acaacaccag tgaataactc ctcacccttc gacatcatgt cgtcgacctg cagactggct
     1261 gtgtataagg gagcctgaca tttatattcc ccagaacatc aggttaatgg cgtttttgat
     1321 gtcattttcg cggtggctga gatcagccac ttcttccccg ataacggaga ccggcacact
     1381 ggccatatcg gtggtcatca tgcgccagct ttcatccccg atatgcacca ccgggtaaag
     1441 ttcacgggag actttatctg acagcagacg tgcactggcc agggggatca ccatccgtcg
     1501 cccgggcgtg tcaataatat cactctgtac atccacaaac agacgataac ggctctctct
     1561 tttataggtg taaaccttaa actgcatttc accagcccct gttctcgtca gcaaaagagc
     1621 cgttcatttc aataaaccgg gcgacctcag ccatcccttc ctgattttcc gctttccagc
     1681 gttcggcacg cagacgacgg gcttcattct gcatggttgt gcttaccaga ccggagatat
     1741 tgacatcata tatgccttga gcaactgata gctgtcgctg tcaactgtca ctgtaatacg
     1801 ctgcttcata gcatacctct ttttgacata cttcgggtat acatatcagt atatattctt
     1861 ataccgcaaa aatcagcgcg caaatacgca tactgttatc tggcttttag taagccggat
     1921 ccacgcgttt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     1981 ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc
     2041 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     2101 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     2161 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     2221 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     2281 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
     2341 accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca
     2401 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
     2461 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
     2521 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
     2581 ttctccattt tagcttcctt agctcctgaa aatctcgacg gatcctaact caaaatccac
     2641 acattatacg agccggaagc ataaagtgta aagacatcat gtcatccggc ggcggcggcg
     2701 ggatatcctc 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
//
