LOCUS       sequence-200982-        3105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobius Assembly Level 1 Acceptor Vector B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3105)
  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 3105)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2018-01-2"; volume: "13(1):e0189892. doi"; pages: " 
            10.1371/journal.pone.0189892. eCollection 2018"
COMMENT     Sequence Label: pMA_Level 1B
FEATURES             Location/Qualifiers
     source          1..3105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      22..79
                     /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(103..120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(274..862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(354..373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1045..1139)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1160..1975)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1160..1975)
                     /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     1883..1902
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2137..2155
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2159..2202)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     promoter        2237..2271
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             2280..2291
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      2980..3051
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3067..3094
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 gtcaatatgc aggtgctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt
       61 ttctttaaaa ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct
      121 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      181 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      241 caggaaccgt aaaaaggccg cgttgctggc gtttttccac aggctccgcc cccctgacga
      301 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      361 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      421 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      481 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      541 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      601 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      661 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      721 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      781 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      841 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      901 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
      961 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1021 ttggtctgac agctcgaggc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     1081 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     1141 aagcgagctc gatatcaaat tagaaaaact catcgagcat caaatgaaac tgcaatttat
     1201 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     1261 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     1321 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     1381 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     1441 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     1501 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
     1561 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     1621 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttccct gggatcgcag
     1681 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     1741 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1801 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cggtagattg
     1861 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1921 tgttggaatt taatcgcggc cttgagcaag acgtttcccg ttgaatatgg ctcattttag
     1981 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     2041 cattatggtg aaagttggaa cctcttacgt gcccgatcaa ctcgagtgcc acctgacgtc
     2101 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat
     2161 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc cacctgcata
     2221 tcagaggaga gagaccttta cggctagctc agtcctaggt atagtgctag ctactagaga
     2281 aagaggagaa atactaaatg tcgcactcaa aacaagcact ggcagatacg atgaaaatga
     2341 cgtggctgat ggaaggttcg gtcaacggtc acgcctttac gattgaaggc gaaggtaccg
     2401 gcaaaccgta tgaaggtaaa cagagtggca cctttcgtgt tacgaaaggc ggtccgctgc
     2461 cgtttgcgtt tgatattgtc gctccgaccc tgaaatacgg tttcaaatgc ttcatgaaat
     2521 acccggcgga tattccggac tactttaaac tggccttccc ggaaggcctg acgtacgatc
     2581 gtaaaattgc gtttgaagac ggcggttgtg cgaccgccac ggtcgaaatg agcctgaagg
     2641 gtaacaccct ggtgcataaa acgaactttc agggcggtaa tttcccgatt gatggtccgg
     2701 tgatgcaaaa acgcaccctg ggctgggaac cgacctctga aaaaatgacg ccgtgcgatg
     2761 gtattatcaa aggcgacacc atcatgtacc tgatggttga aggcggtaaa acgctgaaat
     2821 gtcgttatga aaacaattac cgcgccaaca aaccagtgct gatgccgccg agccactttg
     2881 tggatctgcg tctgacccgc acgaatctgg ataaagaagg tctggcgttc aaactggaag
     2941 aatatgctgt tgcccgtgtg ctggaagtgt aataagcttc aaataaaacg aaaggctcag
     3001 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     3061 cacactggct caccttcggg tgggcctttc tgcgggtctc acgct
//
