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lacI-T5-gusA-pBAV1k vector (V009874#)

Basic Vector Information

Basic Vector Information
Vector Name lacI-T5-gusA-pBAV1k Antibiotic Resistance Kanamycin
Length 6366 bp Type Expression vector
Source Murin CD, Segal K, Bryksin A, Matsumura I.

lacI-T5-gusA-pBAV1k vector Vector Map

t1 repA hypothetical protein hypothetical protein IRII Factor Xa site IRI t0 KanR BioBrick prefix lacIq promoter lacI CAP binding site lac promoter lac operator M13 rev M13 fwd T5 promoter RBS gusA

lacI-T5-gusA-pBAV1k vector Sequence

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LOCUS       Exported                6366 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector lacI-T5-gusA-pBAV1k, complete sequence.
ACCESSION   JF828582
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  AUTHORS   Murin CD, Segal K, Bryksin A, Matsumura I.
  TITLE     Expression Vectors for Acinetobacter baylyi ADP1
  JOURNAL   Appl. Environ. Microbiol. 78 (1), 280-283 (2012)
  PUBMED    22020504
REFERENCE   2  (bases 1 to 6366)
  AUTHORS   Murin CD, Segal K, Bryksin A, Matsumura I.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-2011) Biochemistry, Emory University, 1510 Clifton
            Rd NE, Atlanta, GA 30322, USA
REFERENCE   3  (bases 1 to 6366)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6366
                     /organism="Expression vector lacI-T5-gusA-pBAV1k"
                     /mol_type="other DNA"
                     /db_xref="taxon:1009848"
     regulatory      64..141
                     /regulatory_class="terminator"
                     /note="t1"
     terminator      70..156
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(177..875)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication initiator protein"
                     /protein_id="AEI53594.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(177..875)
                     /gene="repA"
                     /label=repA
     CDS             complement(942..1103)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFC"
                     /protein_id="AEI53595.1"
                     /translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
                     RKESEQKK"
     CDS             complement(1144..1353)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORFB"
                     /protein_id="AEI53596.1"
                     /translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
                     PTFSPTFFINLYLSSFIVVFMITK"
     repeat_region   1306..1354
                     /label=IRIII
                     /note="IRIII"
     repeat_region   1438..1460
                     /label=IRII
                     /note="IRII"
     CDS             1487..1498
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   1507..1638
                     /label=IRI
                     /note="IRI"
     regulatory      1691..1804
                     /regulatory_class="terminator"
                     /note="t0"
     terminator      1700..1794
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1810..2604)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
                     EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             complement(1810..2472)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase APH (3')-IIIA"
                     /label=aminoglycoside phosphotransferase APH
                     /note="APH(3)(5)-IIIa"
                     /protein_id="AEI53597.1"
                     /translation="MTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLL
                     MSEADGVLCSEGYEDEQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLN
                     NDLADVDCENWEEDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFI
                     DLGRSGRADKWYDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    2810..2829
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        2856..2933
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2934..4016
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="AEI53599.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            2934..4016
                     /gene="lacI"
                     /label=lacI
     protein_bind    4029..4050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4065..4095
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4103..4119
                     /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)."
     primer_bind     4127..4143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4159..4175)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4375..4419
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     regulatory      4388..4395
                     /regulatory_class="minus_35_signal"
     misc_feature    4395..4413
                     /label=lac operator
                     /note="lac operator"
     protein_bind    4395..4411
                     /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)."
     regulatory      4413..4453
                     /regulatory_class="ribosome_binding_site"
     regulatory      4413..4418
                     /regulatory_class="minus_10_signal"
     misc_feature    4427..4445
                     /label=lac operator
                     /note="lac operator"
     protein_bind    4427..4443
                     /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)."
     misc_feature    4427
                     /label=+1 (PT5)
                     /note="+1 (PT5)"
     RBS             4466..4477
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4484..6352
                     /codon_start=1
                     /gene="gusA"
                     /product="GusA"
                     /label=gusA
                     /note="6-His beta-D-glucuronidase"
                     /db_xref="ASAP:ABE-0005410"
                     /db_xref="EcoGene:EG11055"
                     /db_xref="UniProtKB/Swiss-Prot:P05804"
                     /protein_id="AEI53598.1"
                     /translation="MGSSHHHHHHSQDPALRAVETPTREIKKLDGLWAFSLDRENCGID
                     QRWWESALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDA
                     VTHYGKVWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVI
                     TDENGKKKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVA
                     NGDVSVELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPL
                     RVGIRSVAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSY
                     RTSHYPYAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQ
                     QAHLQAIKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCV
                     NVMFCDAHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIIT
                     EYGVDTLAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILR
                     VGGNKKGIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQGLCGR"
     gene            4484..6352
                     /gene="gusA"
                     /label=gusA
     CDS             4496..4513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    6365..19
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"

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