pDGO-54 vector (V008005)

LOCUS       40924_14640        6942 bp DNA     circular SYN 18-DEC-2018
DEFINITION  SNAP-XDocII expression vector pDGO-54, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6942)
  AUTHORS   Waller BH, Olson DG, Currie DH, Guss AM, Lynd LR.
  TITLE     Exchange of type II dockerin-containing subunits of the Clostridium 
            thermocellum cellulosome as revealed by SNAP-tags
  JOURNAL   FEMS Microbiol. Lett. 338 (1), 46-53 (2013)
  PUBMED    23082914
REFERENCE   2  (bases 1 to 6942)
  AUTHORS   Olson DG.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-AUG-2012) Thayer School of Engineering, Dartmouth 
            College, Thayer School of Engineering, Dartmouth College, Hanover, 
            NH 03755, USA
REFERENCE   3  (bases 1 to 6942)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6942)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "FEMS 
            Microbiol. Lett."; date: "2013"; volume: "338"; issue: "1"; pages: 
            "46-53"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-AUG-2012) Thayer School of Engineering, Dartmouth College, 
            Thayer School of Engineering, Dartmouth College, Hanover, NH 03755, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(961..1149)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(1672..1693)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(1709..2788)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(2789..2866)
                     /label=lacI promoter
     terminator      3019..3105
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3202..3288
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3385..3471
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3568..3654
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(3754..3797)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        3972..3990
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3991..4015
                     /label=lac operator
                     /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)."
     RBS             4030..4052
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4102..4647
                     /codon_start=1
                     /label=SNAP-tag(R)
                     /note="human O6-alkylguanine-DNA-alkyltransferase variant
                     that forms covalent bonds with benzylguanine derivatives"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     misc_feature    4656..5115
                     /gene="SNAP-XDocII"
                     /label=insert
                     /note="insert"
     CDS             4666..5115
                     /codon_start=1
                     /gene="cipA"
                     /product="dockerin II"
                     /label=cipA
                     /note="cellulosome anchoring protein cohesin subunit;
                     Cthe_3077"
                     /protein_id="AFU51891.1"
                     /translation="MPDFSFDATVAPLVKAGFKVEIVGTELYAVTDANGYFEITGVPAN
                     ASGYTLKISRATYLDRVIANVVVTGDTSVSTSQAPIMMWVGDIVKDNSINLLDVAEVIR
                     CFNATKGSANYVEELDINRNGAINMQDIMIVHKHFGATSSDYDAQ"
     gene            4666..5115
                     /gene="cipA"
                     /label=cipA
     terminator      5205..5252
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5312..5416
                     /label=AmpR promoter
     CDS             5417..6274
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(6319..6832)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"