pIG6-Sia vector (V016213)

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LOCUS       62056_12760        9008 bp DNA     circular SYN 11-AUG-2020
DEFINITION  Cloning vector pIG6-Sia, complete sequence.
ACCESSION   MK353499
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9008)
  AUTHORS   Hu X, Ruan Y, Ding N.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-JAN-2019) Medical College, Dalian University, Xuefu 
            road 10, dalian 116600, China
REFERENCE   2  (bases 1 to 9008)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (01-JAN-2019) Medical College, Dalian University, Xuefu road 10, 
            dalian 116600, China"
FEATURES             Location/Qualifiers
     source          1..9008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..82
                     /label=lacI promoter
     CDS             83..1162
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1178..1199
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1214..1244
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1252..1268
                     /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)."
     primer_bind     1276..1292
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1312..1327
                     /label=ompA RBS
                     /note="ompA RBS"
                     /regulatory_class="ribosome_binding_site"
     sig_peptide     1328..1390
                     /label=OmpA signal peptide
                     /note="signal peptide from the E. coli outer membrane
                     protein OmpA"
     CDS             1736..1753
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      1828..1875
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      1901..2232
                     /note="regulatory region promoter from from pgl locus in 
                     Campylobacter jejuni"
                     /regulatory_class="promoter"
     gene            2236..3780
                     /gene="Delta16psp2,6ST"
                     /label=Delta16psp2,6ST
     CDS             2236..3780
                     /codon_start=1
                     /transl_table=11
                     /gene="Delta16psp2,6ST"
                     /product="protein Delta16psp2,6ST"
                     /label=Delta16psp2,6ST
                     /note="optimized 2,6-sialyltransferase from Vibrionaceae 
                     Photobacterium sp. JT-ISH-224"
                     /protein_id="QEA03482.1"
                     /translation="MKNFLLLTLILLTACNNSEENTQSIIKNDINKTIIDEEYVNLEPI
                     NQSNISFTKHSWVQTCGTQQLLTEQNKESISLSVVAPRLDDDEKYCFDFNGVSNKGEKY
                     ITKVTLNVVAPSLEVYVDHASLPTLQQLMDIIKSEEENPTAQRYIAWGRIVPTDEQMKE
                     LNITSFALINNHTPADLVQEIVKQAQTKHRLNVKLSSNTAHSFDNLVPILKELNSFNNV
                     TVTNIDLYDDGSAEYVNLYNWRDTLNKTDNLKIGKDYLEDVINGINEDTSNTGTSSVYN
                     WQKLYPANYHFLRKDYLTLEPSLHELRDYIGDSLKQMQWDGFKKFNSKQQELFLSIVNF
                     DKQKLQNEYNSSNLPNFVFTGTTVWAGNHEREYYAKQQINVINNAINESSPHYLGNSYD
                     LFFKGHPGGGIINTLIMQNYPSMVDIPSKISFEVLMMTDMLPDAVAGIASSLYFTIPAE
                     KIKFIVFTSTETITDRETALRSPLVQVMIKLGIVKEENVLFWADLPNCETGVCIAV"
     regulatory      3781..3796
                     /label=ompA RBS
                     /note="ompA RBS"
                     /regulatory_class="ribosome_binding_site"
     gene            3797..4837
                     /gene="nueB"
                     /label=nueB
     CDS             3797..4837
                     /codon_start=1
                     /transl_table=11
                     /gene="nueB"
                     /product="NueB"
                     /label=nueB
                     /note="sialic acid synthase from Campylobacter jejuni"
                     /protein_id="QEA03485.1"
                     /translation="MKEIKIQNIIISEEKAPLVVPEIGINHNGSLELAKIMVDAAFSAG
                     AKIIKHQTHIVEDEMSKAAKKVIPGNAKISIYEIMQKCALDYKDELALKEYTEKLGLVY
                     LSTPFSRAGANRLEDMGVSAFKIGSGECNNYPLIKHIAAFKKPMIVSTGMNSIESIKPT
                     VKILLDNEIPFVLMHTTNLYPTPHNLVRLNAMLELKKEFSCMVGLSDHTTDNLACLGAV
                     VLGACVLERHFTDSMHRSGPDIVCSMDTKALKELIIQSEQMAIIRGNNESKKAAKQEQV
                     TIDFAFASVVSIKDIKKGEVLSMDNIWVKRPGLGGISAAEFENILGKKALRDIENDAQL
                     SYEDFA"
     gene            4834..5952
                     /gene="neuC"
                     /label=neuC
     CDS             4834..5952
                     /codon_start=1
                     /transl_table=11
                     /gene="neuC"
                     /product="NeuC"
                     /label=neuC
                     /note="N-acetylglucosamine-6-phosphate 2-epimerase from 
                     Campylobacter jejuni"
                     /protein_id="QEA03484.1"
                     /translation="MKKILFITGSRADYSKIKSLMYRVQNSSEFELYIFATGMHLSKNF
                     GYTVKELYKNGFKNIYEFINYDKYYQTDKALATTIDGFSRYANELKPDLIVVHGDRIEP
                     LAAAIVGALNNILVAHIEGGEISGTIDDSLRHAISKLAHIHLVNDEFAKRRLMQLGEDE
                     KSIFIIGSPDLELLNDNKISLSEAKKYYDINYENYALLMFHPVTTEITSIKNQADNLVK
                     ALIQSNKNYIVIYPNNDLGFELILQSYEEFKNNPRFKLFPSLRFEYFITLLKNADFIIG
                     NSSCILKEALYLKTAGILVGSRQNGRLGNENTLKVNANSDEILKAINTIHKKQDLFSAK
                     LEILDSSKLFFEYLQSGDFFKLSTQKVFKDIK"
     gene            5949..6614
                     /gene="neuA"
                     /label=neuA
     CDS             5949..6614
                     /codon_start=1
                     /transl_table=11
                     /gene="neuA"
                     /product="NeuA"
                     /label=neuA
                     /note="CMP-Neu5Ac synthetase from Campylobacter jejuni"
                     /protein_id="QEA03483.1"
                     /translation="MSLAIIPARGGSKGIKNKNLVLLNNKPLIYYTIKAALNAKSISKV
                     VVSSDSDEILNYAKSQNVDILKRPISLAQDDTTSDKVLLHALKFYKDYEDVVFLQPTSP
                     LRTNIHINEAFNLYKNSNANALISVSECDNKILKAFVCNDCGDLAGICNDEYPFMPRQK
                     LPKTYMSNGAIYILKIKEFLNNPSFLQSKTKHFLMDESSSLDIDCLEDLKKVEQIWKK"
     CDS             6621..6638
                     /label=6xHis
                     /note="6xHis affinity tag"
     rep_origin      6742..7197
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7224..7328
                     /label=AmpR promoter
     CDS             7329..8186
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8360..8948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"