LOCUS Exported 2267 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Provides the Middle-to-End linker 3 which assists Level 2 cloning in Mobius Assembly. ACCESSION . VERSION . KEYWORDS pMA_Auxiliary 3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2267) AUTHORS Andreou AI, Nakayama N TITLE Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly. JOURNAL PLoS One. 2018 Jan 2;13(1):e0189892. doi: 10.1371/journal.pone.0189892. eCollection 2018. PUBMED 29293531 REFERENCE 2 (bases 1 to 2267) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2267 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 2..59 /label=his operon terminator /note="This putative transcriptin terminator from the E. coli his operon has a 2-bp deletion introduced during synthesis. Its efficiency has not been determined." primer_bind complement(83..100) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(254..842) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(334..353) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 1025..1119 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(1140..1955) /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 1863..1882 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2117..2135 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 2139..2182 /label=bacterial terminator /note="putative bacterial transcription terminator" ORIGIN 1 gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat 61 tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 241 cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa aatcgacgct 301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 661 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 901 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 961 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agctcgaggc 1021 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat 1081 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat 1141 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata 1201 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat 1261 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct 1321 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact 1381 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag 1441 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc 1501 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa 1561 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat 1621 tcttctaata cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca 1681 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt 1741 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac 1801 aactctggcg catcgggctt cccatacaat cggtagattg tcgcacctga ttgcccgaca 1861 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc 1921 cttgagcaag acgtttcccg ttgaatatgg ctcattttag cttccttagc tcctgaaaat 1981 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa 2041 cctcttacgt gcccgatcaa ctcgagtgcc acctgacgtc taagaaacca ttattatcat 2101 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt 2161 agctttcgct aaggatgatt tctggaattc cacctgcata tcttgcgcta gagaccgtat 2221 gaggtggatg gtgtgctgcg tggccaaacc catatgcagg tgctgca //