LOCUS       Exported                3982 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD060, complete sequence.
ACCESSION   KM017887
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3982)
  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 3982)
  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 3982)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3982
                     /organism="Broad host range vector vector pCVD060"
                     /mol_type="other DNA"
                     /note="ccdB-Swai-iLOV_S7942 is a reporter device where iLOV
                     is downstream of the ccdB gene and a chloramphenicol 
                     resistance marker, flanked by AhdI restriction sites"
                     /db_xref="taxon:1531918"
     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..1697
                     /note="CVD-cassette; device for the GC-adaptor assembly: 
                     expression/reporter cassette, promoter-reporter module, 
                     custom sequence"
     misc_feature    408..428
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    429..434
                     /label=AgeI
                     /note="AgeI"
     regulatory      454..482
                     /regulatory_class="terminator"
                     /note="lpp T; lpp T; lpp terminator"
     misc_feature    516..523
                     /label=SwaI
                     /note="SwaI"
     regulatory      560..609
                     /regulatory_class="promoter"
                     /note="PccdB; upstream region of the ccdB ORF including 
                     promoter and RBS; derived from the destination cassette 
                     used in the Gateway Technology developed by Life 
                     Technologies"
     CDS             865..1170
                     /codon_start=1
                     /product="ccdB"
                     /label=ccdB
                     /note="from a destination cassette used in the Gateway 
                     Technology developed by Life Technologies"
                     /protein_id="AII99666.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    1211..1218
                     /label=SwaI
                     /note="SwaI"
     misc_feature    1223..1228
                     /label=SacI
                     /note="SacI"
     CDS             1235..1612
                     /codon_start=1
                     /product="iLOV_S7942"
                     /label=iLOV_S7942
                     /note="fluorescent flavoprotein engineered derived from the
                     plant blue-light receptor phototropin; codon optimized for 
                     Synechococcus elongatus PCC7942"
                     /protein_id="AII99665.1"
                     /translation="MIEKNFVITDPRLPDNPIIFASDGFLELTEYSREEILGRNARFLQ
                     GPETDQATVQKIRDAIRDQRETTVQLINYTKSGKKFWNLLHLQPVRDQKGELQYFIGVQ
                     LDGSDHVSGGGGSDYKDDDDK"
     CDS             1586..1609
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    1620..1625
                     /label=SacI
                     /note="SacI"
     regulatory      1635..1663
                     /regulatory_class="terminator"
                     /note="rrnB T2; rrnB T2; rrnB T2 terminator partial 
                     sequence"
     terminator      1635..1662
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    1671..1676
                     /label=XbaI
                     /note="XbaI"
     misc_feature    1677..1697
                     /note="G5C5; GC-adaptor for seamless assembly, flanking 
                     functional devices, custom sequences or cyanobacterial 
                     replicons; synthetic"
     misc_feature    1695..1700
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    complement(1701..1765)
                     /note="lacZa-part1; lacZalpha fragment upstream of EcoICRI"
     misc_feature    1719..1724
                     /label=XbaI
                     /note="XbaI"
     primer_bind     complement(1761..1777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(1766..1888)
                     /regulatory_class="promoter"
                     /note="PlacZa; lacI-lacZ intergenic region derived from 
                     Escherichia coli including the promoter and RBS"
     regulatory      complement(1783..1803)
                     /regulatory_class="other"
                     /note="lacO; LacO; lac operator (lacO) from Invitrogen 
                     pTrcHis2"
     protein_bind    1785..1801
                     /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(1809..1839)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1854..1875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2104..2777)
                     /direction=LEFT
                     /note="ori-pUC19; origin of replication of pUC19"
     rep_origin      complement(2163..2751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2922..3782)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pUC19"
                     /protein_id="AII99664.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3783..3887)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3783..3852)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"
     misc_feature    3913..3918
                     /label=ZraI
                     /note="ZraI"