LOCUS Exported 3040 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Start-Stop Assembly Level 212 plasmid. ACCESSION . VERSION . KEYWORDS pStA212 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3040) AUTHORS Taylor GM, Mordaka PM, Heap JT TITLE Start-Stop Assembly: a functionally scarless DNA assembly system optimized for metabolic engineering. JOURNAL Nucleic Acids Res. 2018 Nov 20. pii: 5193345. doi: 10.1093/nar/gky1182. PUBMED 30462270 REFERENCE 2 (bases 1 to 3040) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3040 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 1..95 /label=lambda t0 terminator /note="transcription terminator from phage lambda" protein_bind 145..166 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 181..211 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 219..235 /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 224..246 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 243..259 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 243..259 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 255..578 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 272..328 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(329..346) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(329..345) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(338..360) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(554..573) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" terminator 607..693 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin 1043..1588 /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 1489..1508 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1705..1722 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(2172..2191) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2183..2998 /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(2256..2275) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" ORIGIN 1 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc 61 tcggttgccg ccgggcgttt tttattggtg agaatgaaga caatgccgga gagagaccca 121 gtagggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 181 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 241 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag 301 aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg 361 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc 421 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg 481 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat 541 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt tccggtctca tactactaaa 601 gtcttcattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc 661 ccagtctttc gactgagcct ttcgttttat ttgatgcccg aaagggcctc gtgatacgca 721 aatctggagc cggtgagcgc ggtatcattg cagcactggg gccagatggt aagccctccc 781 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 841 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 901 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 961 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 1021 accccttaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg 1081 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt 1141 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc 1201 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc 1261 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg 1321 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac 1381 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc 1441 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg gaaacgcctg 1501 gtatctttat agtcctgtcg ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg 1561 cttgtcaggg gggcggagcc tatggaaaaa cggctttgcc gcggccctct cacttccctg 1621 ttaagtatct tcctggcatc ttccaggaaa tctccgcccc gttcgtaagc catttccgct 1681 cgccgcagtc gaacgaccga gcgtagcgag tcagtgagcg aggaagcgga atatatcctg 1741 tatcacatat tctgctgacg caccggtgca gccttttttc tcctgccaca tgaagcactt 1801 cactgacacc ctcatcagtg ccaacatagt aagccagtat acactccgct agcgctgagg 1861 tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc catcatccag 1921 ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc agttggtgat 1981 tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg tgatctgatc 2041 cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg 2101 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat 2161 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgaggcc 2221 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt 2281 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt 2341 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa 2401 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga 2461 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata 2521 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc 2581 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca 2641 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg 2701 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt 2761 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg 2821 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg 2881 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat 2941 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc 3001 ttgggggaaa ttagccgcag ccgaacgacc gagcgccttg //