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vector pCVD028 vector (V001553#)

Basic Vector Information

Basic Vector Information
Vector Name vector pCVD028 Antibiotic Resistance Ampicillin
Length 5612 bp Type Broad host range vector
Source Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.

vector pCVD028 vector Vector Map

lacZa-part2 ZraI ZraI lacZa-part1 PlacZa ori-pUC19 bla AmpR promoter ZraI

vector pCVD028 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5612 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD028, complete sequence.
ACCESSION   KM017916
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5612)
  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 5612)
  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 5612)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5612
                     /organism="Broad host range vector vector pCVD028"
                     /mol_type="other DNA"
                     /note="oribom-sacB-ko is a replication device for E coli 
                     with an origin and a bom site, for conjugal transfer, from 
                     pBR322, carrying a sacB gene for selection of double 
                     recombinants grown with sucrose and that can be assembled 
                     into a double recombination knock-out or knock-in plasmid A
                     point mutation was identified (G249C) in the ORF of sacB 
                     leading to the following amino acid change: E83D (Ryan 
                     Simkovsky) This device appears however functional in 
                     Anabaena sp PCC7120 and Synechococcus elongatus PCC7942"
                     /db_xref="taxon:1531886"
     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=ZraI
                     /note="ZraI"
     misc_feature    complement(408..3327)
                     /note="CVD-ECKO; device for the GC-adaptor assembly: 
                     Escherichia coli origin for KO"
     misc_feature    complement(408..428)
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     Escherichia coli origins for KO; synthetic"
     misc_feature    429..434
                     /label=SacI
                     /note="SacI"
     misc_feature    517..522
                     /label=EcoRV
                     /note="EcoRV"
     CDS             complement(542..1963)
                     /codon_start=1
                     /product="sacB"
                     /label=sacB
                     /note="sacB derived from Bacillus subtilis INSD accession 
                     AAA72307; 1; a point mutation was identified G249C in the 
                     ORF of sacB leading to the following amino acid change: 
                     E83D Ryan Simkovsky; this device appears however functional
                     in Anabaena sp. PCC7120 and Synechococcus elongatus PCC7942
                     Bacillus subtilis"
                     /protein_id="AII99744.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     regulatory      complement(1964..2269)
                     /regulatory_class="promoter"
                     /note="PsacB; derived from Bacillus subtilis INSD accession
                     AAA72307"
     misc_feature    2269..2274
                     /label=SacI
                     /note="SacI"
     oriT            complement(2331..2444)
                     /direction=LEFT
                     /note="bom; bom site of pBR322, for comjugal transfer; 
                     requires a helper plasmid such as pRL623 (or pRK2013)"
     misc_feature    2350..2490
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2617..3290)
                     /direction=LEFT
                     /note="ori-pBR322; origin of replication of pBR322"
     rep_origin      complement(2676..3264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3301..3306
                     /label=MfeI
                     /note="MfeI"
     misc_feature    complement(3307..3327)
                     /note="C3G3; GC-adaptor for seamless assembly, flanking 
                     Escherichia coli origins for KO; synthetic"
     misc_feature    3325..3330
                     /label=ZraI
                     /note="ZraI"
     misc_feature    complement(3331..3395)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    3349..3354
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(3391..3407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3396..3518)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(3413..3433)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    3415..3431
                     /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(3439..3469)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3484..3505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3734..4407)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(3793..4381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4552..5412)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99743.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5413..5517)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(5413..5482)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    5543..5548
                     /label=ZraI
                     /note="ZraI"

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