LOCUS       Exported                5372 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD020, complete sequence.
ACCESSION   KM017921
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5372)
  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 5372)
  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 5372)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5372
                     /organism="Broad host range vector vector pCVD020"
                     /mol_type="other DNA"
                     /note="S7942NS1-TC is a chromosome integration device that 
                     comprises the recombination site NS1 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:1531878"
     misc_feature    46..51
                     /label=ZraI
                     /note="ZraI"
     misc_feature    49..4188
                     /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..855
                     /note="S7942NS1-RA; right arm of the homologous 
                     recombination site NS1 into the chromosome of Synechococcus
                     elongatus PCC7942 derived from INSD accession NC_007604, 
                     U30252"
     rep_origin      866..1539
                     /note="ori-pBR322; origin of replication of pBR322"
     rep_origin      892..1480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1666..1806
                     /label=bom
                     /note="basis of mobility region from pBR322"
     oriT            1712..1825
                     /note="bom; bom site of pBR322, for comjugal transfer; 
                     requires a helper plasmid such as pRL623 (or pRK2013)"
     regulatory      1886..2005
                     /regulatory_class="promoter"
                     /note="Ptet; tet upstream sequence inc; promoter and RBS; 
                     derived from pACYC184 INSD accession X06403"
     promoter        1930..1958
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2006..3196
                     /codon_start=1
                     /product="tet"
                     /label=tet
                     /note="confers tetracycline resistance; tetracycline efflux
                     protein, class C; derived from pACYC184 INSD accession 
                     X06403"
                     /protein_id="AII99752.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    2105..2110
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    2149..2154
                     /label=NheI
                     /note="NheI"
     misc_feature    3197..3286
                     /note="tet-3UTR; 3 UTR of the tetracyclin resistance ORF; 
                     from pACYC184 INSD accession X06403"
     misc_recomb     3291..4161
                     /note="S7942NS1-LA; left arm of the homologous 
                     recombination site NS1 into the chromosome of Synechococcus
                     elongatus PCC7942 derived from INSD accession NC_007604, 
                     U30252"
     misc_feature    3880..3885
                     /label=XbaI
                     /note="XbaI"
     misc_feature    4067..4072
                     /label=NheI
                     /note="NheI"
     misc_feature    4127..4132
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    4162..4167
                     /label=NheI
                     /note="NheI"
     misc_feature    4168..4188
                     /note="GC; GC-adaptor for seamless assembly, flanking BHR 
                     replicons or neutral sites; synthetic"
     misc_feature    4186..4191
                     /label=ZraI
                     /note="ZraI"
     CDS             complement(4312..5172)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99751.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5173..5277)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(5173..5242)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    5303..5308
                     /label=ZraI
                     /note="ZraI"