LOCUS Exported 5882 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR-A-HYG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5882) AUTHORS Garcia-Pedrajas MD, Nadal M, Kapa LB, Perlin MH, Andrews DL, Gold SE TITLE DelsGate, a robust and rapid gene deletion construction method. JOURNAL Fungal Genet Biol. 2008 Apr . 45(4):379-88. PUBMED 18248826 REFERENCE 2 (bases 1 to 5882) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5882 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 335..566 /gene="mutant version of attP" /label=attP1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction (pDONR(TM)201 version)" CDS complement(962..1267) /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind complement(1064..1083) /label=ccdB-fwd /note="ccdB gene, forward primer" CDS complement(1609..2268) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 2183..2202 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(2269..2371) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" protein_bind complement(2516..2747) /gene="mutant version of attP" /label=attP2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction (pDONR(TM)201 version)" primer_bind complement(2860..2879) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2871..3680 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(2938..2957) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3823..4411 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4312..4331 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 4546..4573 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 4665..4751 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" CDS complement(4805..5830) /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" ORIGIN 1 gaagggcgaa cttaagaagg tatgaccggg tcgtccactt accttgcttg acaaacgcac 61 caagttatcg tgcaccaagc agcagatgat aataatgtcc tcgttcctgt ctgctaataa 121 gagtcacact tcgagcgccg ccgctactgc tacaagtggg gctgatctga ccagttgcct 181 aaatgaacca tcttgtcaaa cgacacaaat tttgtgctca ccgcctggac gactaaacca 241 aaataggcat tcattgttga cctccactag ctccagccaa gcccaaaaaa tgctccttca 301 atatcagtta acgctagcat ggatctcggg ccccaaataa tgattttatt ttgactgata 361 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac 421 aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg 481 cataaaaaac agactacata atactgtaaa acacaacata tccagtcact atgaatcaac 541 tacttagatg gtattagtga cctgtagtcg accgacagcc ttccaaatgt tcttcgggtg 601 atgctgccaa cttagtcgac cgacagcctt ccaaatgttc ttctcaaacg gaatcgtcgt 661 atccagccta ctcgctattg tcctcaatgc cgtattaaat cataaaaaga aataagaaaa 721 agaggtgcga gcctcttttt tgtgtgacaa aataaaaaca tctacctatt catatacgct 781 agtgtcatag tcctgaaaat catctgcatc aagaacaatt tcacaactct tatacttttc 841 tcttacaagt cgttcggctt catctggatt ttcagcctct atacttacta aacgtgataa 901 agtttctgta atttctactg tatcgacctg cagactggct gtgtataagg gagcctgaca 961 tttatattcc ccagaacatc aggttaatgg cgtttttgat gtcattttcg cggtggctga 1021 gatcagccac ttcttccccg ataacggaga ccggcacact ggccatatcg gtggtcatca 1081 tgcgccagct ttcatccccg atatgcacca ccgggtaaag ttcacgggag actttatctg 1141 acagcagacg tgcactggcc agggggatca ccatccgtcg cccgggcgtg tcaataatat 1201 cactctgtac atccacaaac agacgataac ggctctctct tttataggtg taaaccttaa 1261 actgcatttc accagtccct gttctcgtca gcaaaagagc cgttcatttc aataaaccgg 1321 gcgacctcag ccatcccttc ctgattttcc gctttccagc gttcggcacg cagacgacgg 1381 gcttcattct gcatggttgt gcttaccaga ccggagatat tgacatcata tatgccttga 1441 gcaactgata gctgtcgctg tcaactgtca ctgtaatacg ctgcttcata gcacacctct 1501 ttttgacata cttcgggtat acatatcagt atatattctt ataccgcaaa aatcagcgcg 1561 caaatacgca tactgttatc tggcttttag taagccggat ccacgcgatt acgccccgcc 1621 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc 1681 acagacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata 1741 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc 1801 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc 1861 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag 1921 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc 1981 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat 2041 tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg 2101 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac 2161 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg 2221 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt 2281 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg 2341 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca 2401 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt 2461 cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttag tcgactacag 2521 gtcactaata ccatctaagt agttgattca tagtgactgg atatgttgtg ttttacagta 2581 ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta 2641 cgtttctcgt tcagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat 2701 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc 2761 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat 2821 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata 2881 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt 2941 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga 3001 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta 3061 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc 3121 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag 3181 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag 3241 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg 3301 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt 3361 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt 3421 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt 3481 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat 3541 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac 3601 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga 3661 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta 3721 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttttcgtt 3781 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 3841 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 3901 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 3961 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 4021 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 4081 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 4141 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 4201 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 4261 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 4321 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 4381 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 4441 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 4501 ggataaccgt attaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc 4561 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac 4621 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg 4681 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt 4741 cgttttattt gatgcctggc agttccctac tctcgcgtta actggttccc ggtcggcatc 4801 tactctattc ctttgccctc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta 4861 cttctacaca gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga 4921 cagtcccggc tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat 4981 cgaaattgcc gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg 5041 cccggaggcg cggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct 5101 gctccataca agccaaccac ggcctccaga agaggatgtt ggcgacctcg tattgggaat 5161 ccccgaacat cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga 5221 cattgttgga gccgaaatcc gcatgcacga ggtgccggac ttcggggcag tcctcggccc 5281 aaagcatcag ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag 5341 tttgccagtg atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt 5401 attgaccgat tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg 5461 cagcgatcgc atccatggcc tccgcgaccg gctggagaac agcgggcagt tcggtttcag 5521 gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat gcaataggtc aggctctcgc 5581 tgaactcccc aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat 5641 aaacataacg atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac 5701 gccctcctac atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt 5761 cggagacgct gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag 5821 gctttttcat ttggatgctt gggtagaata ggtaagtcag attgaatctg aaataaaggg 5881 ag //