LOCUS Exported 2935 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pICH37326 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2935) AUTHORS Engler C, Youles M, Gruetzner R, Ehnert TM, Werner S, Jones JD, Patron NJ, Marillonnet S TITLE A golden gate modular cloning toolbox for plants. JOURNAL ACS Synth Biol. 2014 Nov 21;3(11):839-43. doi: 10.1021/sb4001504. Epub 2014 Feb 20. PUBMED 24933124 REFERENCE 2 (bases 1 to 2935) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2935 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(119..136) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(119..135) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(128..150) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(344..363) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 463..485 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(523..541) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 609..713 /gene="bla" /label=AmpR promoter CDS 933..1724 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 1895..2483 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2384..2403 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2637..2654 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2771..2792 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2807..2837 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2845..2861 /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 2850..2872 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2869..2885 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2869..2885 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 tttggtctca aatggctact caacgaaggg caaaccctag ctctctccat ctaattactg 61 tattctctct gctcgtcgct gtcgtctcag gttgagaccg ggtaccgagc tcgaattcac 121 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc 181 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc 241 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta 301 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg 361 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 421 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 481 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat 541 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg 601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgcgct 721 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt 781 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc 841 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc 901 agggcagtcg ccctaaaaca aagttaaaca tcatgaggga agcggtgatc gccgaagtat 961 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg 1021 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt 1081 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc 1141 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca 1201 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg 1261 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg 1321 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg 1381 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct 1441 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt 1501 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg 1561 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg 1621 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg 1681 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataactgtca gaccaagttt 1741 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga 1801 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag 1861 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa 1921 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag 1981 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg 2041 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat 2101 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta 2161 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg 2221 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc 2281 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa 2341 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc 2401 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt 2461 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct 2521 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 2581 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg 2641 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt 2701 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc 2761 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc 2821 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct 2881 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcccc //