Basic Vector Information
P[acman]-CmR-F-2 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 ℃.
P[acman]-CmR-F-2 vector Sequence
LOCUS 40924_48638 12257 bp DNA circular SYN 18-DEC-2018 DEFINITION BAC cloning vector P[acman]-CmR-F-2, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12257) 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 12257) 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 12257) TITLE Direct Submission REFERENCE 4 (bases 1 to 12257) 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..12257 /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" misc_feature complement(7150..7382) /label=P element 3' end /note="P element 3' end" primer_bind 7389..7405 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 7415..7433 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 7477..7551 /label=multiple cloning site /note="multiple cloning site" promoter complement(7570..7588) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" misc_feature 7624..11639 /label=white dominant eye marker /note="white dominant eye marker" CDS join(8108..8179,8581..8854,8929..9583,9645..9960, 10164..10295,10366..10980) /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(11640..12225) /label=P element 5' end
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