attB-P[acman]-ApR-F-2-5-attB vector (V010107)

Basic Vector Information

      • Vector Name:
      • attB-P[acman]-ApR-F-2-5-attB
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 13067 bp
      • Type:
      • BAC cloning vector
      • Source/Author:
      • Venken KJ, He Y, Hoskins RA, Bellen HJ.

attB-P[acman]-ApR-F-2-5-attB vector Vector Map

attB-P[acman]-ApR-F-2-5-attB13067 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600amplifiable BAC backboneattB integration siteP element 3' endM13 fwdT7 promotermultiple cloning siteT3 promoterwhite dominant eye markerP element 5' end

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 ℃.

attB-P[acman]-ApR-F-2-5-attB vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               13067 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  BAC cloning vector attB-P[acman]-ApR-F-2-5-attB, complete sequence.
ACCESSION   EF106980
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13067)
  AUTHORS   Venken KJ, He Y, Hoskins RA, Bellen HJ.
  TITLE     P[acman]: a BAC transgenic platform for targeted insertion of large 
            DNA fragments in D. melanogaster
  JOURNAL   Science 314 (5806), 1747-1751 (2006)
  PUBMED    17138868
REFERENCE   2  (bases 1 to 13067)
  AUTHORS   Venken KJT., He Y, Hoskins RA, Bellen HJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-NOV-2006) Molecular and Human Genetics, Baylor College
            of Medicine, One Baylor Plaza, Houston, TX 77030, USA
REFERENCE   3  (bases 1 to 13067)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13067
                     /organism="BAC cloning vector
                     attB-P[acman]-ApR-F-2-5-attB"
                     /mol_type="other DNA"
                     /note="available from the Drosophila Genomics Resource 
                     Center: http://dgrc.cgb.indiana.edu/"
                     /db_xref="taxon:412171"
     misc_feature    1..7654
                     /label=amplifiable BAC backbone
                     /note="amplifiable BAC backbone"
     misc_feature    244..717
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(790..1761)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(1761..2936)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    3262..3512
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(3515..4270)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      4361..4580
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      4680..5296
                     /label=OriV
                     /note="OriV"
     rep_origin      4680..5294
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      4771..5302
                     /label=oriV
                     /note="incP origin of replication"
     promoter        6236..6338
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     promoter        6339..6410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6411..7271
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="ABK59034.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    7655..7938
                     /label=attB integration site
                     /note="attB integration site"
     protein_bind    7676..7745
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     misc_feature    7939..8192
                     /label=P element 3' end
                     /note="P element 3' end"
     misc_feature    7960..8192
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     8199..8215
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8225..8243
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    8287..8361
                     /label=multiple cloning site
                     /note="multiple cloning site"
     promoter        complement(8380..8398)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    8434..12449
                     /label=white dominant eye marker
                     /note="white dominant eye marker"
     CDS             join(8918..8989,9391..9664,9739..10393,10455..10770,
                     10974..11105,11176..11790)
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     misc_feature    12450..13056
                     /label=P element 5' end
                     /note="P element 5' end"
     misc_feature    12450..13035
                     /label=P element 5' end

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