LOCUS       sequence-296929-        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(76..867)
                     /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      1115..1703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1604..1623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3225..3249
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    3276..3300
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3352..3386
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             3395..3406
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      4137..4208
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4224..4251
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 acaagaaaag aaaaaacacc cgttagggtg tttttagtta gtgtggcgcg ccgacttgcg
       61 acatgcggtc ctttgttatt tgccgactac cttggtgatc tcgcctttca cgtagtggac
      121 aaattcttcc aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc
      181 ctgtctagct tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc
      241 ggcagcgaca tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa
      301 cgtaagcact acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa
      361 ggtttcattt agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc
      421 cgctggacct accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc
      481 aatgtcgatc gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc
      541 aaattgcagt tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat
      601 ggtgacttct acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag
      661 gtcgttgatc aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa
      721 atcaatatca ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc
      781 cagcaacgtc ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga
      841 tacttcggcg atcaccgctt ccctcatgat gtttaacttt gttttagggc gactgccctg
      901 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
      961 ctgcttggat gcccgaggca tagactgtac cccaaaaaaa cagtcataac aagccatgaa
     1021 aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttcgatcgg attggcggtt
     1081 atgcggttcc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1141 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1201 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1261 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1321 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1381 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1441 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1501 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1561 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1621 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1681 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1741 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1801 gtattaccgc ctttgagtga gctgccgatc cgccgccagt tcaggccgtc cggccccgga
     1861 ttctgaccat aatttcatcg aaaaaagggg cgcagccctt cttgttctaa tagttctata
     1921 agttcaggcg aaaatcgtgc agcaattaca aaaggttgcg cgtctataag tggggaatcc
     1981 agccgcaaaa gtggggaatc cagccgcaaa agtggggaat ccagccgaaa tcgcggattg
     2041 acgagtggga tttcccgccc aataaatcca ccatgggaaa gacactcgac gttgcccgcg
     2101 accgggcctt tgaccagacc gcgaccgtgc tgcccgccga aatggcgcgg ggggtctata
     2161 tgcgcaacgc ccccagcctc gcggccctga agctgatgca tctgatgatc gccacggcgg
     2221 gcgggcgcat ggccgatgac gtgcgccacg aaatgcggct ggccgacatc cgcaagatcg
     2281 acggtatgga taaccacacc cgggccagcc tgaccccgct ctttgcggaa ctggcggcgg
     2341 cggtgctgac ccacgacgac ccggaaaagc gggtcgtgac catcggtggc ctgctggacg
     2401 aagcccggat cgattaccgg cacgaggtca gcggcgatct tctggtgtcg tggaccttcc
     2461 gcagcatgtt ccgccgcatg gcggcggaat cgaaccattg ggcgattctc gaccggcaga
     2521 ccgtgttcca cctcggcagc aagtattccg tgctgttgtt ccagcacatc gccagcttca
     2581 aggaatacga ccacattacc ggcaagacct ttaccgtgcc ggagttgcgg gctgtgtttg
     2641 gtatccccga gggcaaaatc aagcgtttcg cagacctcaa cagagacgtg ctgacgcccg
     2701 ccattgccga aatcaaccag ctttcccgcc tgactctgac cgccacgccg aacaagatcg
     2761 ggcgcaccgt ggccagcgtg acgattgctt gggaagaaaa gcccctcgaa ggcaagcgct
     2821 cgaccaaggc cgaactggac cgcccgaagg tgggccggaa ggcccggcgc gacggcaccg
     2881 ccgagacggt ggcacgggcc ttcccggcat cgggcgggat cgagttcgac cagcattggc
     2941 gcgacctcaa gcgggcggcg ggctgcaaca tggacaacac catgatcgcc gacaaattcc
     3001 gggcatggtg cgccgggaag ggcctcgctc tcgatgcccg gaacatcgaa caggcgttca
     3061 gcagcttctg caccaaggtg ggccgggtct gaccatcatc agttcggtgg tcttccgacg
     3121 aacaataagg ccgcaaatcg cggccttttt tattgataac aaaaccggct cagttctgcg
     3181 tagaaaccaa catgcaagct ccaccgggtg caaagcggca gcggcggcag gatatattca
     3241 attgtaaatg gcttcatgtc cgggaaatct actggtggca ggatatattg tggtgtaaac
     3301 aatggagaaa aagattctgg aattcggtct cacagaggag atatgcaggt gtttacggct
     3361 agctcagtcc taggtatagt gctagctact agagaaagag gagaaatact aaatgcgtaa
     3421 aggcgaagaa ctgttcacgg gcgtagttcc gattctggtc gagctggacg gcgatgtgaa
     3481 cggtcataag tttagcgttc gcggtgaagg tgagggcgac gcgaccaacg gcaaactgac
     3541 cctgaagttc atctgcacca ccggtaaact gccggtgcct tggccgacct tggtgacgac
     3601 gttgacgtat ggcgtgcagt gttttgcgcg ttatccggac cacatgaaac aacacgattt
     3661 cttcaaatct gcgatgccgg agggttacgt ccaggagcgt accatttcct tcaaggatga
     3721 tggctactac aaaactcgcg cagaggttaa gtttgaaggt gacacgctgg tcaatcgtat
     3781 cgaattgaag ggtatcgact ttaaagagga tggtaacatt ctgggccata aactggagta
     3841 taacttcaac agccataatg tttacattac ggcagacaag caaaagaacg gcatcaaggc
     3901 caatttcaag attcgccaca atgttgagga cggtagcgtc caactggccg accattacca
     3961 gcagaacacc ccaattggtg acggtccggt tttgctgccg gataatcact atctgagcac
     4021 ccaaagcgtg ctgagcaaag atccgaacga aaaacgtgat cacatggtcc tgctggaatt
     4081 tgtgaccgct gcgggcatca cccacggtat ggacgagctg tataagtaat gagcttcaaa
     4141 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     4201 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gcacctgcat
     4261 atacccctgc aggaattgtt gattttgtga tgactgatgg caggatatat gcggttgtaa
     4321 ttcattttta ttgtctaaat ttctgtattt gtttgtttgt tcggttgtaa atttttttgg
     4381 aaga
//
