pTD1-phiC31Int2 vector (V002760)

Basic Vector Information

Vector Name:
pTD1-phiC31Int2
Antibiotic Resistance:
Ampicillin
Length:
4989 bp
Type:
Expression vector
Replication origin:
ori
Source/Author:
Yonemura N, Tamura T, Uchino K, Kobayashi I, Tatematsu K, Iizuka T, Sezutsu H, Muthulakshmi M, Nagaraju J, Kusakabe T.

pTD1-phiC31Int2 vector Map

pTD1-phiC31Int24989 bp6001200180024003000360042004800T7 promoter5'UTRattB1NcoI site was replaced with CCGTGGphage phi-C31 integraseattB23'UTRsynthetic poly(A)20 regionT7 terminatorM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

pTD1-phiC31Int2 vector Sequence

LOCUS       40924_42944        4989 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTD1-phiC31Int2 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  AUTHORS   Yonemura N, Tamura T, Uchino K, Kobayashi I, Tatematsu K, Iizuka T, 
            Sezutsu H, Muthulakshmi M, Nagaraju J, Kusakabe T.
  TITLE     PhiC31 integrase-mediated cassette exchange in silkworm embryos
  JOURNAL   Mol. Genet. Genomics 287 (9), 731-739 (2012)
  PUBMED    22842670
REFERENCE   2  (bases 1 to 4989)
  AUTHORS   Yonemura N, Kobayashi I, Uchino K, Sezutsu H, Tamura T, Kusakabe T.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-APR-2012) Contact:Naoyuki Yonemura National Institute 
            of Agrobiological Sciences (NIAS), MAFF; Ohwashi 1-2, Tsukuba 
            Science City 305-8634, Japan
REFERENCE   3  (bases 1 to 4989)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4989)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Genet.
            Genomics"; date: "2012"; volume: "287"; issue: "9"; pages: "731-739"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-APR-2012) Contact:Naoyuki Yonemura National Institute of 
            Agrobiological Sciences (NIAS), MAFF; Ohwashi 1-2, Tsukuba Science 
            City 305-8634, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        240..258
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           259..304
                     /label=polyhedrin 5' UTR
                     /note="5' UTR from the Malacosoma neustria
                     nucleopolyhedrovirus polyhedrin gene (Ezure et al., 2006)"
     protein_bind    323..347
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    387..392
                     /label=NcoI site was replaced with CCGTGG
                     /note="NcoI site was replaced with CCGTGG"
     CDS             407..2221
                     /codon_start=1
                     /label=phage phi-C31 integrase
                     /note="integrase from phage phi-C31"
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     protein_bind    complement(2230..2254)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     3'UTR           2296..2668
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     misc_feature    2673..2694
                     /label=synthetic poly(A)20 region
                     /note="synthetic poly(A)20 region"
     terminator      2695..2742
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2768..2784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2792..2808)
                     /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(2816..2846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2861..2882)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3170..3758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3932..4789)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4790..4894)
                     /label=AmpR promoter

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