LOCUS       Exported                3772 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD026, complete sequence.
ACCESSION   KM017914
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3772)
  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 3772)
  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 3772)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3772
                     /organism="Broad host range vector vector pCVD026"
                     /mol_type="other DNA"
                     /note="oribom is a replication device for E coli with an 
                     origin and a bom site, for conjugal transfer, from pBR322 
                     to assembled with a cyanobacterial replicon"
                     /db_xref="taxon:1531884"
     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..1487
                     /note="CVD-ECcya; device for the GC-adaptor assembly: 
                     Escherichia coli origin to be assembled with a 
                     cyanobacterial replicon"
     misc_feature    408..428
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     cyanobacterial replicons or Escherichia coli origins to be 
                     assembled with a cyanobacterial replicon; synthetic"
     misc_feature    429..434
                     /label=MfeI
                     /note="MfeI"
     rep_origin      445..1118
                     /note="ori-pBR322; origin of replication of pBR322"
     rep_origin      471..1059
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1245..1385
                     /label=bom
                     /note="basis of mobility region from pBR322"
     oriT            1291..1404
                     /note="bom; bom site of pBR322, for comjugal transfer; 
                     requires a helper plasmid such as pRL623 (or pRK2013)"
     misc_feature    1461..1466
                     /label=NheI
                     /note="NheI"
     misc_feature    1467..1487
                     /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    1485..1490
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    complement(1491..1555)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    1509..1514
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(1551..1567)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(1556..1678)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(1573..1593)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    1575..1591
                     /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(1599..1629)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1644..1665
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1894..2567)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(1953..2541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2712..3572)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99741.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3573..3677)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3573..3642)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    3703..3708
                     /label=ZraI
                     /note="ZraI"