LOCUS       sequence-319270-        4384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobius Assembly cloning and expression vector for plants with 
            high-copy number origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4384)
  AUTHORS   Andreou , Andreas I., Nirkko, Jessica , Ochoa-Villarreal, Marisol , 
            Nakayama, Naomi
  TITLE     Mobius Assembly for Plant Systems highlights promoter-terminator 
            interaction in gene regulation
  JOURNAL   bioRxiv, March 31, 2021
REFERENCE   2  (bases 1 to 4384)
  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: "bioRxiv, 
            March 31, 2021"
COMMENT     Sequence Label: pMAP L2B
FEATURES             Location/Qualifiers
     source          1..4384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(99..890)
                     /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      1138..1726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1627..1646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3248..3272
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    3299..3323
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3375..3409
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             3418..3429
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4160..4231
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4247..4274
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 cggttgtaaa tttttttgga agaacaagaa aagaaaaaac acccgttagg gtgtttttag
       61 ttagtgtggc gcgccgactt gcgacatgcg gtcctttgtt atttgccgac taccttggtg
      121 atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
      181 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
      241 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact
      301 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg
      361 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga
      421 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct
      481 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga
      541 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga
      601 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca
      661 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc
      721 cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact
      781 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca
      841 actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat gatgtttaac
      901 tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctccataa catcaaacat
      961 cgacccacgg cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg taccccaaaa
     1021 aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc
     1081 aaggttcgat cggattggcg gttatgcggt tcccgtagaa aagatcaaag gatcttcttg
     1141 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1201 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1261 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1321 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1381 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1441 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1501 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     1561 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1621 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     1681 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     1741 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     1801 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgccg atccgccgcc
     1861 agttcaggcc gtccggcccc ggattctgac cataatttca tcgaaaaaag gggcgcagcc
     1921 cttcttgttc taatagttct ataagttcag gcgaaaatcg tgcagcaatt acaaaaggtt
     1981 gcgcgtctat aagtggggaa tccagacgca aaagtgggga atccagccgc aaaagtgggg
     2041 aatccagccg aaatcgcgga ttgacgagtg ggatttcccg cccaataaat ccaccatggg
     2101 aaagacactc gacgttgccc gcgaccgggc ctttgaccag accgcgaccg tgctgcccgc
     2161 cgaaatggcg cggggggtct atatgcgcaa cgcccccagc ctcgcggccc tgaagctgat
     2221 gcatctgatg atcgccacgg cgggcgggcg catggccgat gacgtgcgcc acgaaatgcg
     2281 gctggccgac atccgcaaga tcgacggtat ggataaccac acccgggcca gcctgacccc
     2341 gctctttgcg gaactggcgg cggcggtgct gacccacgac gacccggaaa agcgggtcgt
     2401 gaccatcggt ggcctgctgg acgaagcccg gatcgattac cggcacgagg tcagcggcga
     2461 tcttctggtg tcgtggacct tccgcagcat gttccgccgc atggcggcgg aatcgaacca
     2521 ttgggcgatt ctcgaccggc agaccgtgtt ccacctcggc agcaagtatt ccgtgctgtt
     2581 gttccagcac atcgccagct tcaaggaata cgaccacatt accggcaaga cctttaccgt
     2641 gccggagttg cgggctgtgt ttggtatccc cgagggcaaa atcaagcgtt tcgcagacct
     2701 caacagagac gtgctgacgc ccgccattgc cgaaatcaac cagctttccc gcctgactct
     2761 gaccgccacg ccgaacaaga tcgggcgcac cgtggccagc gtgacgattg cttgggaaga
     2821 aaagcccctc gaaggcaagc gctcgaccaa ggccgaactg gaccgcccga aggtgggccg
     2881 gaaggcccgg cgcgacggca ccgccgagac ggtggcacgg gccttcccgg catcgggcgg
     2941 gatcgagttc gaccagcatt ggcgcgacct caagcgggcg gcgggctgca acatggacaa
     3001 caccatgatc gccgacaaat tccgggcatg gtgcgccggg aagggcctcg ctctcgatgc
     3061 ccggaacatc gaacaggcgt tcagcagctt ctgcaccaag gtgggccggg tctgaccatc
     3121 atcagttcgg tggtcttccg acgaacaata aggccgcaaa tcgcggcctt ttttattgat
     3181 aacaaaaccg gctcagttct gcgtagaaac caacatgcaa gctccaccgg gtgcaaagcg
     3241 gcagcggcgg caggatatat tcaattgtaa atggcttcat gtccgggaaa tctactggtg
     3301 gcaggatata ttgtggtgta aacaatggag aaaaagattc tggaattcgg tctcacagag
     3361 gagatatgca ggtgtttacg gctagctcag tcctaggtat agtgctagct actagagaaa
     3421 gaggagaaat actaaatgcg taaaggcgaa gaactgttca cgggcgtagt tccgattctg
     3481 gtcgagctgg acggcgatgt gaacggtcat aagtttagcg ttcgcggtga aggtgagggc
     3541 gacgcgacca acggcaaact gaccctgaag ttcatctgca ccaccggtaa actgccggtg
     3601 ccttggccga ccttggtgac gacgttgacg tatggcgtgc agtgttttgc gcgttatccg
     3661 gaccacatga aacaacacga tttcttcaaa tctgcgatgc cggagggtta cgtccaggag
     3721 cgtaccattt ccttcaagga tgatggctac tacaaaactc gcgcagaggt taagtttgaa
     3781 ggtgacacgc tggtcaatcg tatcgaattg aagggtatcg actttaaaga ggatggtaac
     3841 attctgggcc ataaactgga gtataacttc aacagccata atgtttacat tacggcagac
     3901 aagcaaaaga acggcatcaa ggccaatttc aagattcgcc acaatgttga ggacggtagc
     3961 gtccaactgg ccgaccatta ccagcagaac accccaattg gtgacggtcc ggttttgctg
     4021 ccggataatc actatctgag cacccaaagc gtgctgagca aagatccgaa cgaaaaacgt
     4081 gatcacatgg tcctgctgga atttgtgacc gctgcgggca tcacccacgg tatggacgag
     4141 ctgtataagt aatgagcttc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     4201 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     4261 tgggcctttc tgcgcacctg catatacccc tgcaggaatt gttgattttg tgatgactga
     4321 tggcaggata tatgcggttg taattcattt ttattgtcta aatttctgta tttgtttgtt
     4381 tgtt
//
