LOCUS       Exported                3771 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD027, complete sequence.
ACCESSION   KM017915
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3771)
  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 3771)
  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 3771)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3771
                     /organism="Broad host range vector vector pCVD027"
                     /mol_type="other DNA"
                     /note="oribom-ko is a replication device for E coli with an
                     origin and a bom site, for conjugal transfer, from pBR322 
                     to be assembled into a double recombination knock-out or 
                     knock-in vector"
                     /db_xref="taxon:1531885"
     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    404..409
                     /label=ZraI
                     /note="ZraI"
     misc_feature    407..1486
                     /note="CVD-ECKO; device for the GC-adaptor assembly: 
                     Escherichia coli origin for KO"
     misc_feature    407..427
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     Escherichia coli origins for KO; synthetic"
     misc_feature    428..433
                     /label=MfeI
                     /note="MfeI"
     rep_origin      444..1117
                     /note="ori-pBR322; origin of replication of pBR322"
     rep_origin      470..1058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1244..1384
                     /label=bom
                     /note="basis of mobility region from pBR322"
     oriT            1290..1403
                     /note="bom; bom site of pBR322, for comjugal transfer; 
                     requires a helper plasmid such as pRL623 (or pRK2013)"
     misc_feature    1460..1465
                     /label=SacI
                     /note="SacI"
     misc_feature    1466..1486
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     Escherichia coli origins for KO; synthetic"
     misc_feature    1484..1489
                     /label=ZraI
                     /note="ZraI"
     misc_feature    complement(1490..1554)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    1508..1513
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(1550..1566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(1555..1677)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(1572..1592)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    1574..1590
                     /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(1598..1628)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1643..1664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1893..2566)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(1952..2540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2711..3571)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99742.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3572..3676)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3572..3641)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    3702..3707
                     /label=ZraI
                     /note="ZraI"