LOCUS Exported 2679 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMLLCK-K112603 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2679) TITLE J.C. Anderson Lab Plasmids REFERENCE 2 (bases 1 to 2679) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2679 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 6..29 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(59..76) /label=L4440 /note="L4440 vector, forward primer" protein_bind complement(138..162) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(312..331) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 323..1138 /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(396..415) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" CDS 1348..2007 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1414..1433) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" terminator 2028..2122 /label=lambda t0 terminator /note="transcription terminator from phage lambda" rep_origin 2158..2514 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" protein_bind 2576..2600 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" ORIGIN 1 gatctgacta caaggatgac gacgacaagt aaggatccta actcgctcct caggcttcct 61 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 121 aggcggtaat caattcgacc cagctttctt gtacaaagtg gttgatcgga tctgctgatc 181 cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg 241 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat 301 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctccaggcc 361 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt 421 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt 481 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa 541 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga 601 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata 661 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc 721 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca 781 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg 841 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt 901 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg 961 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg 1021 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat 1081 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc 1141 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggaggatct 1201 ctgaagactc tcgagctgca ggactcacag ctcctcggat ctttgatcgg gcacgtaaga 1261 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg 1321 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt 1381 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt 1441 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat 1501 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg 1561 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt 1621 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac 1681 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg 1741 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg 1801 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc 1861 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt 1921 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac 1981 tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat 2041 agatccagta atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg 2101 ggcgtttttt attggtgaga atccaagcct cggatctagc agttcaacct gttgatagta 2161 cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat 2221 gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatggctaa cgtactaagc 2281 tctcatgttt cacgtactaa gctctcatgt ttgaacaata aaattaatat aaatcagcaa 2341 cttaaatagc ctctaaggtt ttaagtttta taagaaaaaa aagaatatat aaggctttta 2401 aagcttttaa ggtttaacgg ttgtggacaa caagccaggg atgtaacgca ctgagaagcc 2461 cttagagcct ctcaaagcaa ttttgagtga cacaggaaca cttaacggct gacatgggaa 2521 ttagccatgg gcccgtgcga atcacggatc tcccgatcaa gaattgccat tatcaacaag 2581 tttgtacaaa aaagcaggct ccgaattggt atcacgaggc agaatttcag ataaaaaaaa 2641 tccttagctt tcgctaagga tctgaagtgg aattcatga //