LOCUS       Exported                4004 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD065, complete sequence.
ACCESSION   KM017881
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4004)
  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 4004)
  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 4004)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4004
                     /organism="Broad host range vector vector pCVD065"
                     /mol_type="other DNA"
                     /note="aadA_nt is a spectinomycin and streptomycin (SpSm) 
                     resistance device About hundred nts of the omega cassette 
                     follow the CDS but there is not a synthetic terminator This
                     device works well"
                     /db_xref="taxon:1531923"
     misc_feature    complement(146..404)
                     /note="lacZa-part2; lacZalpha fragment upstream of EcoICRI"
     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..1719
                     /note="CVD-ANTR; device for the GC-adaptor assembly: 
                     antibiotic resistance markers"
     misc_feature    408..428
                     /note="GC; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers or Escherichia coli origins 
                     to be assembled with a cyanobacterial replicon; synthetic"
     misc_feature    429..434
                     /label=NheI
                     /note="NheI"
     regulatory      435..818
                     /regulatory_class="promoter"
                     /note="PaadA; upstream region of aadA ORF including 
                     promoter and RBS; derived from pRL1383a in INSD accession 
                     AF403426, originally derived from the omega interposon INSD
                     accession M60473"
     CDS             819..1610
                     /codon_start=1
                     /product="aadA"
                     /label=aadA
                     /note="confers spectinomycin and/or streptomycin 
                     resistance; aminoglycoside-3:adenyltransferase fusion; 
                     derived from pRL1383a in INSD accession AF403426, 
                     originally derived from the omega interposon INSD accession
                     M60473"
                     /protein_id="AII99650.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    1693..1698
                     /label=AgeI
                     /note="AgeI"
     misc_feature    1699..1719
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    1717..1722
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    complement(1723..1787)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    1741..1746
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(1783..1799)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(1788..1910)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(1805..1825)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    1807..1823
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(1831..1861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1876..1897
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2126..2799)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(2185..2773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2944..3804)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99651.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3805..3909)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3805..3874)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    3935..3940
                     /label=ZraI
                     /note="ZraI"