LOCUS       Exported                5124 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD023, complete sequence.
ACCESSION   KM017924
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5124)
  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 5124)
  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 5124)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5124
                     /organism="Broad host range vector vector pCVD023"
                     /mol_type="other DNA"
                     /note="S7942NS2-TC is a chromosome integration device that 
                     comprises the recombination site NS2 for Synechococcus 
                     elongatus PCC7942, an origin of replication for E coli and 
                     a bom site for conjugal transfer, from pBR322, and a 
                     Tetracycline resistance marker"
                     /db_xref="taxon:1531881"
     misc_feature    46..51
                     /label=ZraI
                     /note="ZraI"
     misc_feature    49..3940
                     /note="CVD-BHR-NS; device for the GC-adaptor assembly: BHR 
                     replicon and neutral sites or right arm of a double 
                     recombination KO"
     misc_feature    49..69
                     /note="G5C5; GC-adaptor for seamless assembly, flanking a 
                     BHR replicons, neutral sites, or cyanobacterial replicons; 
                     synthetic"
     misc_feature    70..75
                     /label=XbaI
                     /note="XbaI"
     misc_recomb     76..536
                     /note="S7942NS2-RA; right arm of the homologous 
                     recombination site NS2 into the chromosome of Synechococcus
                     elongatus PCC7942 derived from INSD accession NC_007604, 
                     U44761"
     misc_feature    445..450
                     /label=SacI
                     /note="SacI"
     rep_origin      547..1220
                     /note="ori-pBR322; origin of replication of pBR322"
     rep_origin      573..1161
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1347..1487
                     /label=bom
                     /note="basis of mobility region from pBR322"
     oriT            1393..1506
                     /note="bom; bom site of pBR322, for comjugal transfer; 
                     requires a helper plasmid such as pRL623 (or pRK2013)"
     misc_feature    complement(1567..1656)
                     /note="tet-3UTR; 3 UTR of the tetracyclin resistance ORF; 
                     from pACYC184 INSD accession X06403"
     CDS             complement(1657..2847)
                     /codon_start=1
                     /product="tet"
                     /label=tet
                     /note="confers tetracycline resistance; tetracycline efflux
                     protein, class C; derived from pACYC184 INSD accession 
                     X06403"
                     /protein_id="AII99756.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    2699..2704
                     /label=NheI
                     /note="NheI"
     misc_feature    2743..2748
                     /label=EcoRV
                     /note="EcoRV"
     regulatory      complement(2848..2967)
                     /regulatory_class="promoter"
                     /note="Ptet; tet upstream sequence inc; promoter and RBS; 
                     derived from pACYC184 INSD accession X06403"
     promoter        complement(2895..2923)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     misc_recomb     2972..3913
                     /note="S7942NS2-LA; left arm of the homologous 
                     recombination site NS2 into the chromosome of Synechococcus
                     elongatus PCC7942 derived from INSD accession NC_007604, 
                     U44761"
     misc_feature    3914..3919
                     /label=NheI
                     /note="NheI"
     misc_feature    3920..3940
                     /note="GC; GC-adaptor for seamless assembly, flanking BHR 
                     replicons or neutral sites; synthetic"
     misc_feature    3938..3943
                     /label=ZraI
                     /note="ZraI"
     CDS             complement(4064..4924)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99755.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4925..5029)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(4925..4994)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    5055..5060
                     /label=ZraI
                     /note="ZraI"