pBAC-Lac vector (V009334)

Basic Vector Information

Vector Name:
pBAC-Lac
Antibiotic Resistance:
Chloramphenicol
Length:
7964 bp
Type:
Cloning vector
Replication origin:
ori2
Source/Author:
Asakawa S, Abe I, Kudoh Y, Kishi N, Wang Y, Kubota R, Kudoh J, Kawasaki K, Minoshima S, Shimizu N.

pBAC-Lac vector Map

pBAC-Lac7964 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800MCSM13 revlac operatorlac promoterCAP binding siteSP6 promoterCmRcat promoterori2repEincCsopAsopBsopCcosloxPT7 promoterM13 fwd

pBAC-Lac vector Sequence

LOCUS       40924_5529        7964 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBAC-Lac DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7964)
  AUTHORS   Asakawa S, Abe I, Kudoh Y, Kishi N, Wang Y, Kubota R, Kudoh J, 
            Kawasaki K, Minoshima S, Shimizu N.
  TITLE     Human BAC library: construction and rapid screening
  JOURNAL   Gene 191 (1), 69-79 (1997)
  PUBMED    9210591
REFERENCE   2  (bases 1 to 7964)
  AUTHORS   Asakawa S, Shimizu N.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2001) Nobuyoshi Shimizu, Keio University School of
            Medicine, Department of Molecular Biology; 35 Shinanomachi, 
            Shinjuku-ku, Tokyo 160-8582, Japan 
            (E-mail:nshimizu@dmb.med.keio.ac.jp, Tel:81-3-5363-3755, 
            Fax:81-3-3351-2370)
REFERENCE   3  (bases 1 to 7964)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7964)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Gene"; 
            date: "1997"; volume: "191"; issue: "1"; pages: "69-79"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-AUG-2001) Nobuyoshi Shimizu, Keio University School of Medicine,
            Department of Molecular Biology"; volume: " 35 Shinanomachi, 
            Shinjuku-ku, Tokyo 160-8582, Japan 
            (E-mail:nshimizu@dmb.med.keio.ac.jp, Tel:81-3-5363-3755, Fax"; 
            pages: "81-3-3351-2370"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..57)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(67..83)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(91..107)
                     /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)."
     promoter        complement(115..145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(160..181)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        complement(378..395)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(539..1195)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1196..1298)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      2225..2444
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             2535..3287
                     /codon_start=1
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    3293..3543
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             3869..5041
                     /codon_start=1
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             5044..6012
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    6088..6561
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    complement(6821..7219)
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(7237..7270)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        7286..7304
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7945..7961
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"

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