pDONR221-P1P4载体

LOCUS Exported 4774 bp ds-DNA circular SYN 10-AUG-2016DEFINITION synthetic circular DNAKEYWORDS pDONR221-P1P4 AUTHORS Biofeng Lab.FEATURES Location/Qualifiers source 1..4774 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 268..295 /note="rrnB T2 terminator" /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /note="rrnB T1 terminator" /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 537..553 /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" protein_bind 570..801 /gene="mutant version of attP" /bound_moiety="BP Clonase(TM)" /note="attP1" /note="recombination site for the Gateway(R) BP reaction (pDONR(TM)201 version)" CDS complement(1197..1502) /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /note="ccdB" /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" CDS complement(1852..2505) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG" promoter complement(2506..2608) /note="cat promoter" /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" protein_bind complement(2753..2984) /gene="mutant version of attP" /bound_moiety="BP Clonase(TM)" /note="attP4" /note="recombination site for the Gateway(R) BP reaction" promoter complement(3016..3034) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3039..3055) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" CDS 3168..3977 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /note="KanR" /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 4124..4712 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa 721 aaaacagact acataatact gtaaaacaca acatatccag tcactatgaa tcaactactt 781 agatggtatt agtgacctgt agtcgaccga cagccttcca aatgttcttc gggtgatgct 841 gccaacttag tcgaccgaca gccttccaaa tgttcttctc aaacggaatc gtcgtatcca 901 gcctactcgc tattgtcctc aatgccgtat taaatcataa aaagaaataa gaaaaagagg 961 tgcgagcctc ttttttgtgt gacaaaataa aaacatctac ctattcatat acgctagtgt 1021 catagtcctg aaaatcatct gcatcaagaa caatttcaca actcttatac ttttctctta 1081 caagtcgttc ggcttcatct ggattttcag cctctatact tactaaacgt gataaagttt 1141 ctgtaatttc tactgtatcg acctgcagac tggctgtgta taagggagcc tgacatttat 1201 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca 1261 gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc 1321 cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc 1381 agacgtgcac tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat aatatcactc 1441 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc 1501 atttcaccag cccctgttct cgtcagcaaa agagccgttc atttcaataa accgggcgac 1561 ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac gacgggcttc 1621 attctgcatg gttgtgctta ccagaccgga gatattgaca tcatatatgc cttgagcaac 1681 tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata cctctttttg 1741 acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat 1801 acgcatactg ttatctggct tttagtaagc cggatccacg cggcgtttac gccccccctg 1861 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca 1921 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata 1981 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa 2041 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt 2101 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa 2161 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg 2221 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc 2281 catacggaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata 2341 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt 2401 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca 2461 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc 2521 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg 2581 aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg 2641 cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac 2701 aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttagtcg actacaggtc 2761 actaatacca tctaagtagt tgattcatag tgactggata tgttgtgttt tacagtatta 2821 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt 2881 ttctcgttca acttttctat acaaagttgg cattataaga aagcattgct tatcaatttg 2941 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagg 3001 gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc 3061 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat 3121 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc 3181 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata 3241 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc 3301 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag 3361 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt 3421 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat 3481 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt 3541 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat 3601 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg 3661 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc 3721 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg 3781 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc 3841 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc 3901 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc 3961 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc 4021 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt 4081 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 4141 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 4201 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 4261 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 4321 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 4381 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 4441 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 4501 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 4561 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 4621 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 4681 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt