LOCUS Exported 2762 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCM5.47 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2762) TITLE Seydoux Lab - Third Generation Vector Kit REFERENCE 2 (bases 1 to 2762) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2762 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind complement(624..748) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(871..966) /gene="mutant version of attL" /label=attL3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1003..1022) /label=T7 /note="T7 promoter, forward primer" promoter complement(1004..1022) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1027..1043) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1027..1043) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1145..1164) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1156..1965 /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(1223..1242) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2112..2700 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2601..2620 /label=pBR322ori-F /note="pBR322 origin, forward primer" 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 tcgggccctg cagctctaga gctcgaattc tacaggtcac 601 taataccatc taagtagttg attcatagtg actgcatatg ttgtgtttta cagtattatg 661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt 721 ctcgttcaac tttcttgtac aaagtgggat aattttgccg tattttccat attttgtttt 781 gtatatttat ccactcaccc cctctctttg tcctgtgaat gaacttgtgc caaaaaagcc 841 aaaaataatt tacttttttt aaagacatat caactttatt atacaaagtt ggcattataa 901 aaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca 961 ttatttggag ctccatggta gcgttaacgc ggccgcgata tcccctatag tgagtcgtat 1021 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac 1081 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt 1141 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc 1201 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt 1261 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc 1321 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa 1381 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc 1441 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt 1501 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt 1561 aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat 1621 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa 1681 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt 1741 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt 1801 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt 1861 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat 1921 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat 1981 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca 2041 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc 2101 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 2161 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2221 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 2281 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 2341 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 2401 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 2461 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg 2521 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2581 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2641 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2701 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 2761 tt //