attB-P[acman]-CmR-F-2-attB vector (V010106)

Basic Vector Information

Vector Name:
attB-P[acman]-CmR-F-2-attB
Antibiotic Resistance:
Chloramphenicol
Length:
12525 bp
Type:
BAC cloning vector
Replication origin:
ori2
Source/Author:
Venken KJ, He Y, Hoskins RA, Bellen HJ.

attB-P[acman]-CmR-F-2-attB vector Vector Map

attB-P[acman]-CmR-F-2-attB12525 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000sopCsopBsopAincCrepEori2oriVcat promoterCmRattBP 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]-CmR-F-2-attB vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_320       12525 bp DNA     circular SYN 17-DEC-2018
DEFINITION  BAC cloning vector attB-P[acman]-CmR-F-2-attB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12525)
  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 12525)
  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 12525)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12525)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science"; 
            date: "2006"; volume: "314"; issue: "5806"; pages: "1747-1751"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-NOV-2006) Molecular and Human Genetics, Baylor College of 
            Medicine, One Baylor Plaza, Houston, TX 77030, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(244..717)
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(793..1761)
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(1764..2936)
                     /codon_start=1
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    complement(3262..3512)
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(3518..4270)
                     /codon_start=1
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      complement(4361..4580)
                     /direction=LEFT
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      4659..5372
                     /label=oriV
                     /note="incP origin of replication"
     promoter        6236..6338
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6339..6995
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     protein_bind    7134..7203
                     /label=attB
                     /note="attB site for the phi-C31 integrase (Groth et al.,
                     2000)"
     misc_feature    complement(7418..7650)
                     /label=P element 3' end
                     /note="P element 3' end"
     primer_bind     7657..7673
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7683..7701
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    7745..7819
                     /label=multiple cloning site
                     /note="multiple cloning site"
     promoter        complement(7838..7856)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    7892..11907
                     /label=white dominant eye marker
                     /note="white dominant eye marker"
     CDS             join(8376..8447,8849..9122,9197..9851,9913..10228,
                     10432..10563,10634..11248)
                     /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    complement(11908..12493)
                     /label=P element 5' end

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