LOCUS Exported 2432 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION POC12356, MetClo assembly vector with p15A replication origin and chloramphenicol resistance for BsaI-based MetClo, adaptor sequence type a-b. ACCESSION . VERSION . KEYWORDS pMXLC_aFb SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2432) AUTHORS Lin D, O'Callaghan CA TITLE MetClo: methylase-assisted hierarchical DNA assembly using a single type IIS restriction enzyme. JOURNAL Nucleic Acids Res. 2018 Jul 9. pii: 5050631. doi: 10.1093/nar/gky596. PUBMED 29986052 REFERENCE 2 (bases 1 to 2432) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2432 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 111..132 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 147..177 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 185..201 /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)." primer_bind 190..212 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 209..225 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 209..225 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 221..544 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 238..294 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(295..312) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(295..311) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(304..326) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(520..539) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 819..921 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 922..1581 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(988..1007) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" rep_origin 1607..2152 /direction=RIGHT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind 2053..2072 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2269..2286 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cactctgtga cttggtctca aggtggagtg agaccggcct agcaggtgtg agtttgcagc 61 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt 121 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt 181 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag 241 cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga attcactggc 301 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc 361 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc 421 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca 481 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc 541 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 601 gctcccggca tccgcttaca gacaagctgt gacaagtacc acctggaacg ccaaaggtct 661 cacgcttgcc tgagaccaag tcacgaagtg tgcggccgcc ttcagcggcc gcacgaataa 721 atacctgtga cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc 781 gggaagccct gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc 841 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt 901 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata 961 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat 1021 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac 1081 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggagttc 1141 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc 1201 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc 1261 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat 1321 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc 1381 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg 1441 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat 1501 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat 1561 gagtggcagg gcggggcgta agagcctgtt taataagatg atcttcttga gatcgttttg 1621 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga 1681 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac 1741 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta 1801 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca 1861 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag 1921 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg 1981 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag 2041 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg 2101 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt 2161 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg 2221 ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg 2281 agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt 2341 tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca 2401 gtatacactc cgctagcgcg caagcggccg ca //