LOCUS       Exported                6942 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  SNAP-XDocII expression vector pDGO-54, complete sequence.
ACCESSION   JX500710
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6942
                     /organism="SNAP-XDocII expression vector pDGO-54"
                     /mol_type="other DNA"
                     /label=Escherichia coli
                     /note="Escherichia coli"
                     /db_xref="taxon:1238570"
     rep_origin      1..589
                     /label=pMB1 origin
                     /note="pMB1 origin"
     misc_feature    complement(958..1149)
                     /label=rop pMB1
                     /note="rop pMB1"
     CDS             complement(958..1149)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    complement(1706..2788)
                     /label=LacI from Escherichia coli
                     /note="LacI from Escherichia coli"
     CDS             complement(1706..2788)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(2789..2866)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      3019..3105
                     /gene="Escherichia coli rrnB"
                     /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      3751..3837
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      3872..3989
                     /regulatory_class="promoter"
                     /note="T7"
     promoter        3972..3990
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3991..4015
                     /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)."
     protein_bind    3992..4016
                     /label=lac operator binding site
                     /bound_moiety="lac operator"
     RBS             4030..4052
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     gene            4045..5115
                     /gene="SNAP-XDocII"
                     /label=SNAP-XDocII
     regulatory      4045..4051
                     /regulatory_class="ribosome_binding_site"
                     /gene="SNAP-XDocII"
     CDS             4060..5115
                     /codon_start=1
                     /gene="SNAP-XDocII"
                     /product="SNAP-XDocII fusion protein"
                     /label=SNAP-XDocII
                     /protein_id="AFU51890.1"
                     /translation="MASTMDIKLTGEFAMDKDCEMKRTTLDSPLGKLELSGCEQGLHEI
                     KLLGKGTSAADAVEVPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQ
                     QESFTRQVLWKLLKVVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVV
                     SSSGAVGGYEGGLAVKEWLLAHEGHRLGKPGLGPAGGYILPDFSFDATVAPLVKAGFKV
                     EIVGTELYAVTDANGYFEITGVPANASGYTLKISRATYLDRVIANVVVTGDTSVSTSQA
                     PIMMWVGDIVKDNSINLLDVAEVIRCFNATKGSANYVEELDINRNGAINMQDIMIVHKH
                     FGATSSDYDAQ"
     misc_feature    4060..4655
                     /gene="SNAP-XDocII"
                     /label=SNAP protein
                     /note="SNAP protein"
     CDS             4102..4647
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5417..6277
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR; ampicillin-resistance protein"
                     /protein_id="AFU51892.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5417..6277
                     /gene="bla"
                     /label=bla
     rep_origin      complement(6319..6832)
                     /direction=LEFT
                     /label=M13 origin
                     /note="M13 origin"