载体基本信息 | |||
载体名称 | pBAC-TS | 载体抗性 | Tetracycline |
---|---|---|---|
载体长度 | 10475 bp | 载体类型 | Cloning vector |
载体来源 | Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA, Del Vecchio D, Collins JJ. |
pBAC-TS 载体载体图谱
pBAC-TS 载体载体序列
LOCUS Exported 10475 bp ds-DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector pBAC-TS, complete sequence. ACCESSION KX264178 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10475) AUTHORS Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA, Del Vecchio D, Collins JJ. TITLE Creating Single-Copy Genetic Circuits JOURNAL Mol. Cell 63 (2), 329-336 (2016) PUBMED 27425413 REFERENCE 2 (bases 1 to 10475) AUTHORS Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA, Del Vecchio D, Collins JJ. TITLE Direct Submission JOURNAL Submitted (18-MAY-2016) Biological Engineering, MIT, 45 Carleton St, MIT Building E25-Rm302, Cambridge, MA 02142, USA REFERENCE 3 (bases 1 to 10475) AUTHORS . TITLE Direct Submission COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..10475 /organism="Cloning vector pBAC-TS" /mol_type="other DNA" /db_xref="taxon:1883154" CDS complement(102..1073) /codon_start=1 /gene="sopB" /product="partitioning protein for the bacterial F plasmid" /label=sopB /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP" CDS complement(1073..2248) /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 2574..2824 /gene="incC" /label=incC /note="incompatibility region of the bacterial F plasmid" CDS complement(2827..3582) /codon_start=1 /gene="repE" /product="replication initiation protein for the bacterial F plasmid" /label=repE /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM RLSYIEKKKGRQTTHIVFSFRDITSMTTG" rep_origin 3673..3892 /label=ori2 /note="secondary origin of replication for the bacterial F plasmid; also known as oriS" rep_origin 3674..3891 /label=OriS origin /note="OriS origin" rep_origin 3962..4586 /label=OriV origin /note="OriV origin" rep_origin 3968..4582 /label=oriV /note="origin of replication for the bacterial F plasmid" promoter 5440..5542 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 5543..6202 /codon_start=1 /product="CamR" /label=CamR /protein_id="ANW61907.1" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" regulatory 6331..6436 /regulatory_class="terminator" /note="T0 transcriptional terminator from phage lambda" terminator 6337..6431 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(6491..7207) /codon_start=1 /product="green fluorescent protein mut3b" /label=green fluorescent protein mut3b /note="gfp(mut3b)" /protein_id="ANW61911.1" /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" regulatory complement(7259..7332) /regulatory_class="promoter" /note="PLtetO" promoter complement(7259..7332) /label=PLtetO-1 promoter /note="modified phage lambda PL promoter with tet operator sites (Lutz and Bujard, 1997)" prim_transcript 7278 /label=transcription start /note="transcription start" protein_bind 7289..7307 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 7314..7332 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" CDS complement(7375..8538) /codon_start=1 /product="LacI" /label=LacI /protein_id="ANW61910.1" /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQAANKNEENTNEVPTFMLNAGQANYLSQ" misc_feature complement(7375..7458) /label=mfLon-specific tag pdt2 /note="mfLon-specific tag pdt2" regulatory complement(8575..8648) /regulatory_class="promoter" /note="PLtetO" promoter complement(8575..8648) /label=PLtetO-1 promoter /note="modified phage lambda PL promoter with tet operator sites (Lutz and Bujard, 1997)" prim_transcript 8594 /label=transcription start /note="transcription start" protein_bind 8605..8623 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 8630..8648 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" regulatory 8667..8741 /regulatory_class="promoter" /note="Ptrc-2" promoter 8680..8709 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" prim_transcript 8715 /label=transcription start /note="transcription start" protein_bind 8717..8733 /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)." CDS 8780..9403 /codon_start=1 /product="TetR" /label=TetR /protein_id="ANW61909.1" /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" regulatory 9446..9520 /regulatory_class="promoter" /note="Ptrc-2" promoter 9459..9488 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 9496..9512 /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)." prim_transcript 9496 /label=transcription start /note="transcription start" CDS 9563..10273 /codon_start=1 /product="mCherry" /label=mCherry /protein_id="ANW61908.1" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" regulatory 10328..10432 /regulatory_class="terminator" /note="T1 transcriptional terminator from rrnB operon" terminator 10333..10419 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene"
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