LOCUS       Exported                3493 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD035, complete sequence.
ACCESSION   KM017910
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3493)
  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 3493)
  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 3493)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3493
                     /organism="Broad host range vector vector pCVD035"
                     /mol_type="other DNA"
                     /note="PpsbAI-LTRBS-YFP is a functional device where YFP is
                     driven by the PpsbAI promoter isolated from Synechococcus 
                     elongatus PCC7942; RBS and flanking sequences from the 
                     pENTR/SD/D-Topo vector"
                     /db_xref="taxon:1531893"
     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..1498
                     /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      428..618
                     /regulatory_class="promoter"
                     /note="PpsbAI_S7942; upstream region of psbA1 up to 25nt 
                     upstream of the start codon in Synechococcus elongatus 
                     PCC7942"
     regulatory      625..665
                     /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             636..656
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             666..1385
                     /codon_start=1
                     /product="yfp"
                     /label=yfp
                     /note="genetic mutant of green fluorescent protein, derived
                     from Aequorea victoria; its excitation peak is 514nm and 
                     its emission peak is 527nm"
                     /protein_id="AII99734.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVRFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(1402..1426)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1436..1464
                     /regulatory_class="terminator"
                     /note="rrnB T2; rrnB T2; rrnB T2 terminator partial 
                     sequence"
     terminator      1436..1463
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    1472..1477
                     /label=XbaI
                     /note="XbaI"
     misc_feature    1478..1498
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    1496..1501
                     /label=EcoRV
                     /note="EcoRV"
     rep_origin      complement(1687..2360)
                     /direction=LEFT
                     /note="colE1_pMA-RQ; origin of replication of colE1 found 
                     inderived from pMA-RQ GeneART vector"
     rep_origin      complement(1746..2334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2505..3365)
                     /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="AII99735.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3366..3470)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3366..3435)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"