LOCUS       sequence-317046-        4592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for plants with low-copy number origin of 
            replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4592)
  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 4592)
  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: pLXMA_RK2 L2A
FEATURES             Location/Qualifiers
     source          1..4592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    172..196
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    223..247
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        290..324
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             333..344
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1075..1146
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1162..1189
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1397..2188)
                     /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"
     CDS             complement(2460..3608)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      4001..4532
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 aaaaaaagga tctcaagaag atcctttgat cttttctacc catcatcagt tcggtggtct
       61 tccgacgaac aaaaaggccg caaatcgcgg ccttttttat tgataacaaa accggctcag
      121 ttctgcgtag aaaccaacat gcaagctcca ccgggtgcaa agcggcagcg gcggcaggat
      181 atattcaatt gtaaatggct tcatgtccgg gaaatctact ggtggcagga tatattgtgg
      241 tgtaaacaat ggagaaaaag agaattcggt ctcaggagat atgcaggtgt ttacggctag
      301 ctcagtccta ggtatagtgc tagctactag agaaagagga gaaatactaa atgcgtaaag
      361 gcgaagaact gttcacgggc gtagttccga ttctggtcga gctggacggc gatgtgaacg
      421 gtcataagtt tagcgttcgc ggtgaaggtg agggcgacgc gaccaacggc aaactgaccc
      481 tgaagttcat ctgcaccacc ggtaaactgc cggtgccttg gccgaccttg gtgacgacgt
      541 tgacgtatgg cgtgcagtgt tttgcgcgtt atccggacca catgaaacaa cacgatttct
      601 tcaaatctgc gatgccggag ggttacgtcc aggagcgtac catttccttc aaggatgatg
      661 gctactacaa aactcgcgca gaggttaagt ttgaaggtga cacgctggtc aatcgtatcg
      721 aattgaaggg tatcgacttt aaagaggatg gtaacattct gggccataaa ctggagtata
      781 acttcaacag ccataatgtt tacattacgg cagacaagca aaagaacggc atcaaggcca
      841 atttcaagat tcgccacaat gttgaggacg gtagcgtcca actggccgac cattaccagc
      901 agaacacccc aattggtgac ggtccggttt tgctgccgga taatcactat ctgagcaccc
      961 aaagcgtgct gagcaaagat ccgaacgaaa aacgtgatca catggtcctg ctggaatttg
     1021 tgaccgctgc gggcatcacc cacggtatgg acgagctgta taagtaatga gcttcaaata
     1081 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1141 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgc acctgcatat
     1201 accctgcagg aattgttgat tttgtgatga ctgatggcag gatatatgcg gttgtaattc
     1261 atttttattg tctaaatttc tgtatttgtt tgtttgttcg gttgtaaatt tttttggaag
     1321 aacaagaaaa gaaaaaacac ccgttagggt gtttttagtt agtgtggcgc gccgacttgc
     1381 gacatgcggt cctttgttat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga
     1441 caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag
     1501 cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt
     1561 cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca
     1621 acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta
     1681 aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg
     1741 ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat
     1801 caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc
     1861 caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa
     1921 tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa
     1981 ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca
     2041 aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg
     2101 ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg
     2161 atacttcggc gatcaccgct tccctcatga tgtttaactt tgttttaggg cgactgccct
     2221 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
     2281 gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcataa caagccatga
     2341 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttcgatcg gattggcggt
     2401 tatgcggttg cgatgctggt ggctgctgaa cccccagccg gaactgaccc cacaaggccc
     2461 tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag gccgctgcct
     2521 cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc
     2581 gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg gtgcgagctg
     2641 aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc ccatatgaat
     2701 ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag gacctggcaa
     2761 cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga caccgattcc
     2821 aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat
     2881 tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg gcaaagctcg
     2941 tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc caagacctgc
     3001 tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg
     3061 tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg
     3121 cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc
     3181 cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt cgtgtgtttc
     3241 agcaacgcgg cctgcttggc ttcgctgacc tgttttgcca ggtcctcgcc ggcggttttt
     3301 cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc
     3361 gcctcctgtt cgagacgacg cgaacgctcc acggcggccg atggcgcggg cagggcaggg
     3421 ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac tatcgagccg
     3481 acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca
     3541 tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc
     3601 cgattcattc accctccttg cgggattgcc ccggaattaa ttccccggat cgatccgtcg
     3661 atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag tttactttgc
     3721 agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg cttgctgtcc
     3781 ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct
     3841 ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc
     3901 accgtttctg cggactggct ttctacgtgg ctgccatttt tggggtgagg ccgttcgcgg
     3961 ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg gagggttcga
     4021 gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta
     4081 aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta gcggtggccg
     4141 aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag cccctcaaat
     4201 gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc
     4261 cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact tatccccagg
     4321 cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag
     4381 gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt caacgccgcg
     4441 ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt gagggccaag
     4501 ttttccgcga ggtatccaca acgccggcgg ccctacatgg ctctgctgta gtgagtgggt
     4561 tgcgctccgg cagcggtcct gatcccccgc ag
//
