LOCUS       40924_49097        4474 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD017, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4474)
  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 4474)
  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 4474)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4474)
  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..4474
                     /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="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    429..434
                     /label=AgeI
                     /note="AgeI"
     regulatory      454..482
                     /note="lpp T; lpp T; lpp terminator"
                     /regulatory_class="terminator"
     regulatory      513..613
                     /note="PnrsB; nickel inducible promoter; derived from 
                     Synechocystis sp. PCC6803 and used in the pSyn1 vector 
                     developed by Life Technologies"
                     /regulatory_class="promoter"
     promoter        665..695
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             749..1405
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     regulatory      1445..1494
                     /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             1750..2052
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     terminator      2127..2154
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    2163..2168
                     /label=XbaI
                     /note="XbaI"
     misc_feature    2169..2189
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    2187..2192
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    2211..2216
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(2253..2269)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2277..2293)
                     /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(2301..2331)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2346..2367)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2655..3243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3417..4274)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4275..4379)
                     /label=AmpR promoter
     misc_feature    4405..4410
                     /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 ccgccgccgc
      421 cgccggatac cggttaatag tacctgtgaa gtgaaaaatg gcgcacattg tgcgacattt
      481 ttttctcgag caggtagtcg gcaaataacc ctcggtctga tcttagcggg ggaaggagat
      541 tttcacctga atttcatacc ccctttggca gactgggaaa atcttggaca aattcccaat
      601 ttgaggtggt gtgaagcttc gaattcgaag gagcccttgt ttaaacgcat taggcacccc
      661 aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc
      721 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
      781 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
      841 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
      901 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
      961 ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     1021 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     1081 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     1141 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     1201 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     1261 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     1321 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     1381 ctgcgatgag tggcagggcg gggcgtaaac gcgtggatcc ggcttactaa aagccagata
     1441 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
     1501 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
     1561 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
     1621 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
     1681 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
     1741 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
     1801 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
     1861 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     1921 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     1981 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     2041 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgaccatag
     2101 gtttaaacac aaagtggtcc tgcaggagaa ggccatcctg acggatggcc tttttgaagc
     2161 tttctagaat cgggggcccc cggggggata tcctcggtac ccggggatcc tctagagtcg
     2221 acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2281 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2341 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2401 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2461 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2521 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2581 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2641 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2701 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2761 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2821 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2881 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2941 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3001 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3061 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3121 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3181 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3241 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3301 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3361 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3421 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3481 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3541 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3601 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3661 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3721 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3781 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3841 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3901 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3961 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4021 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4081 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4141 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     4201 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     4261 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     4321 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     4381 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     4441 tataaaaata ggcgtatcac gaggcccttt cgtc
//
