Basic Vector Information
- Vector Name:
- pESPBAC
- Antibiotic Resistance:
- Apramycin
- Length:
- 13791 bp
- Type:
- Expression vector
- Replication origin:
- ori2
- Source/Author:
- Huang H, Shang G.
pESPBAC vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pESPBAC vector Sequence
LOCUS 40924_17626 13791 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pESPBAC, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13791)
AUTHORS Huang H, Shang G.
TITLE Direct Submission
JOURNAL Submitted (12-MAY-2008) College of Life Sciences, No.1 Wenyuan Rd.,
XiXia District, Nanjing, Jiangsu Province 210046, P.R.China
REFERENCE 2 (bases 1 to 13791)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13791)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(12-MAY-2008) College of Life Sciences, No.1 Wenyuan Rd., XiXia
District, Nanjing, Jiangsu Province 210046, P.R.China"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13791
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 289..305
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 312..330
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(390..406)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 414..430
/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)."
promoter complement(438..468)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 483..504
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(792..1380)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1554..2411)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2412..2516)
/label=AmpR promoter
primer_bind 2990..3006
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3082..3100)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3118..3134)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3142..3158)
/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(3166..3196)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3211..3232)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
oriT complement(3447..3556)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
CDS complement(3794..3805)
/codon_start=1
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
/translation="IEGR"
protein_bind 3876..3975
/label=phage phi-C31 attP
/note="attachment site of phage phi-C31"
CDS 3985..5799
/codon_start=1
/label=phage phi-C31 integrase
/note="integrase from phage phi-C31"
/translation="VDTYAGAYDRQSRERENLSAASPATQRSANEDKAADLQREVERDG
GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
EKRASITWAKPPTDDDEDDAQDGTEDVAA"
CDS 5896..6696
/codon_start=1
/label=ApmR
/note="aminoglycoside 3-N-acetyltransferase type IV"
/translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
CDS complement(7062..7706)
/codon_start=1
/product="catabolite gene activator"
/label=catabolite gene activator
/note="Crp"
/protein_id="ACD75468.1"
/translation="MVASALPAKIKNIDKLLVHCQRRRYTAKSNIICAGDRAETLSFII
KGSVTILIEDDDGHEMIIAYLNHGDFFGELGLFEPVDGEQQRSAWVRAKTECEVAEISY
EKFRELARQDPEILYALGSQMAQRLRNTTRKVGDLAFFDVTGRVARCLLELCKQPDAMT
HPDGMQIKITRQEIGRIVGCSREMVGRVLKDLEERSLVQVKGKTMVVHGTR"
promoter complement(7706..7808)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin 8735..8958
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 9049..9801
/codon_start=1
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
/translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
misc_feature 9807..10057
/gene="incC"
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS 10383..11555
/codon_start=1
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
/translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
CDS 11558..12526
/codon_start=1
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
/translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
misc_feature 12602..13075
/label=sopC
/note="centromere-like partitioning region of the bacterial
F plasmid"
misc_feature complement(13335..13733)
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"
protein_bind complement(13751..13784)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
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