LOCUS       40924_49282        3982 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD060, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3982)
  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 3982)
  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 3982)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3982)
  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..3982
                     /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"
     misc_feature    516..523
                     /label=SwaI
                     /note="SwaI"
     regulatory      560..609
                     /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             865..1167
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     misc_feature    1211..1218
                     /label=SwaI
                     /note="SwaI"
     misc_feature    1223..1228
                     /label=SacI
                     /note="SacI"
     CDS             1238..1555
                     /label=miniSOG
                     /note="mini singlet oxygen generator, a fluorescent
                     flavoprotein suitable for correlated light and electron 
                     microscopy (Shu et al., 2011)"
     CDS             1586..1609
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1620..1625
                     /label=SacI
                     /note="SacI"
     terminator      1635..1662
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     misc_feature    1671..1676
                     /label=XbaI
                     /note="XbaI"
     misc_feature    1677..1697
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    1695..1700
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    1719..1724
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(1761..1777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1785..1801)
                     /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(1809..1839)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1854..1875)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2163..2751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2925..3782)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3783..3887)
                     /label=AmpR promoter
     misc_feature    3913..3918
                     /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 gccaataacc cagggattta aatacgcgtg gatccggctt
      541 actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata
      601 tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca
      661 gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg
      721 gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag
      781 cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg
      841 ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa aagagagagc
      901 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg
      961 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
     1021 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
     1081 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
     1141 gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca cagccagtct
     1201 gcaggtcgac atttaaatga aggagctctg cagcatgatc gaaaagaact ttgtgatcac
     1261 cgatcctcgc ctgcccgata accccatcat ctttgccagt gatggctttc tggaactgac
     1321 cgaatacagt cgcgaagaaa tcctgggtcg caacgctcgc tttctgcaag gccctgaaac
     1381 cgatcaggcc accgtgcaga aaatccgcga cgccatccgc gatcagcgcg aaaccaccgt
     1441 gcagctgatc aactacacca aaagcggcaa aaagttttgg aacctgctgc atctgcaacc
     1501 cgtgcgcgat cagaaaggcg aactgcaata ctttatcggc gtgcagctgg atggcagcga
     1561 tcacgtgagt ggcggtggcg gtagcgatta caaagatgat gatgataagt agaaatacag
     1621 agctccccgg caggagaagg ccatcctgac ggatggcctt tttgaagctt tctagaatcg
     1681 ggggcccccg gggggatatc ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat
     1741 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1801 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     1861 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1921 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1981 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2041 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2101 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2161 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2221 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2281 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2341 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2401 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2461 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2521 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2581 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2641 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2701 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2761 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2821 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2881 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2941 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3001 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3061 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3121 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3181 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3241 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3301 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3361 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3421 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3481 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3541 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3601 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3661 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3721 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3781 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3841 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3901 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3961 cgtatcacga ggccctttcg tc
//
