LOCUS       Exported                3334 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD033, complete sequence.
ACCESSION   KM017908
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3334)
  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 3334)
  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 3334)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3334
                     /organism="Broad host range vector vector pCVD033"
                     /mol_type="other DNA"
                     /note="PconII-LTRBS-eCFP is a functional device where eCFP 
                     is driven by a PconII promoter; RBS and flanking sequences 
                     from the pENTR/SD/D-Topo vector"
                     /db_xref="taxon:1531891"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    389..394
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    392..1339
                     /note="CVD-cassette; device for the GC-adaptor assembly: 
                     expression/reporter cassette, promoter-reporter module, 
                     custom sequence"
     misc_feature    392..412
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    413..418
                     /label=AgeI
                     /note="AgeI"
     regulatory      425..459
                     /regulatory_class="promoter"
                     /note="PconII; -35 to +1 region derived from conII 
                     synthetic promoter; synthetic"
     promoter        425..453
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      466..506
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS_LT; RBS and flanking sequences found in Life 
                     Tech pENTR-SD/D Topo, start downstream of the att site and 
                     includes the T7 gene enhancer, RBS, TOPO binding site and 
                     directional TOPO overhang; Life Technologies; synthetic"
     RBS             477..497
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             507..1226
                     /codon_start=1
                     /product="eCFP"
                     /label=eCFP
                     /note="genetic mutant of green fluorescent protein, derived
                     from Aequorea victoria; its excitation peak is 440nm and 
                     its emission peak is 475nm; Aequorea victoria modified"
                     /protein_id="AII99730.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(1243..1267)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1277..1305
                     /regulatory_class="terminator"
                     /note="rrnB T2; rrnB T2; rrnB T2 terminator partial 
                     sequence"
     terminator      1277..1304
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    1313..1318
                     /label=XbaI
                     /note="XbaI"
     misc_feature    1319..1339
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    1337..1342
                     /label=EcoRV
                     /note="EcoRV"
     rep_origin      complement(1528..2201)
                     /direction=LEFT
                     /note="colE1_pMA-RQ; origin of replication of colE1 found 
                     inderived from pMA-RQ GeneART vector"
     rep_origin      complement(1587..2175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2346..3206)
                     /codon_start=1
                     /product="bla_pMA-RQ"
                     /label=bla_pMA-RQ
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pMA-RQ GeneART vector, has one point mutation 
                     compared to pUC19"
                     /protein_id="AII99731.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3207..3311)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3207..3276)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"