LOCUS       sequence-317042-        4500 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 4500)
  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 4500)
  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 L1A
FEATURES             Location/Qualifiers
     source          1..4500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    124..148
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    175..199
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        241..275
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             284..295
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      984..1055
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1071..1098
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1329..2144)
                     /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"
     primer_bind     2136..2155
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(2320..3468)
                     /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      3861..4392
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gttcggtggt cttccgacga acaataaggc cgcaaatcgc ggcctttttt attgataaca
       61 aaaccggctc agttctgcgt agaaaccaac atgcaagctc caccgggtgc aaagcggcag
      121 cggcggcagg atatattcaa ttgtaaatgg cttcatgtcc gggaaatcta ctggtggcag
      181 gatatattgt ggtgtaaaca atggagaaaa agagaattca cctgcatatg gagagagacc
      241 tttacggcta gctcagtcct aggtatagtg ctagctacta gagaaagagg agaaatacta
      301 aatgtcgcac tcaaaacaag cactggcaga tacgatgaaa atgacgtggc tgatggaagg
      361 ttcggtcaac ggtcacgcct ttacgattga aggcgaaggt accggcaaac cgtatgaagg
      421 taaacagagt ggcacctttc gtgttacgaa aggcggtccg ctgccgtttg cgtttgatat
      481 tgtcgctccg accctgaaat acggtttcaa atgcttcatg aaatacccgg cggatattcc
      541 ggactacttt aaactggcct tcccggaagg cctgacgtac gatcgtaaaa ttgcgtttga
      601 agacggcggt tgtgcgaccg ccacggtcga aatgagcctg aagggtaaca ccctggtgca
      661 taaaacgaac tttcagggcg gtaatttccc gattgatggt ccggtgatgc aaaaacgcac
      721 cctgggctgg gaaccgacct ctgaaaaaat gacgccgtgc gatggtatta tcaaaggcga
      781 caccatcatg tacctgatgg ttgaaggcgg taaaacgctg aaatgtcgtt atgaaaacaa
      841 ttaccgcgcc aacaaaccag tgctgatgcc gccgagccac tttgtggatc tgcgtctgac
      901 ccgcacgaat ctggataaag aaggtctggc gttcaaactg gaagaatatg ctgttgcccg
      961 tgtgctggaa gtgtaataag cttcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1021 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     1081 cgggtgggcc tttctgcggg tctcacgctc agaatatgca ggtgcctgca ggaattgttg
     1141 attttgtgat gactgatggc aggatatatg cggttgtaat tcatttttat tgtctaaatt
     1201 tctgtatttg tttgtttgtt cggttgtaaa tttttttgga agaacaagaa aagaaaaaac
     1261 acccgttagg gtgtttttag ttagtgtggc gcgccgactt gcgacatgcg gtcctttgca
     1321 atcaactatt agaaaaattc atccagcatc agatgaaatt gcagtttgtt catatccgga
     1381 ttatcaatgc catatttctg aaacagacgt ttttgcaggc tcgggctaaa ttcgcccagg
     1441 cagttccaca gaatggccag atcctgataa cgatccgcaa tgcccacacg gcccacatca
     1501 atgcagccaa tcagtttgcc ttcatcgaaa atcaggttat ccaggctaaa atcgccgtgg
     1561 gtcaccacgc tatccgggct aaacggcagc agtttatgca tttctttcca cacctgttcc
     1621 accggccagc cgttacgttc atcatcaaaa tcgctcgcat ccaccaggcc gttgttcata
     1681 cggctctgcg cctgggccag acgaaacaca cgatcgctgt taaacgggca gttgcacacc
     1741 ggaatgctat gcagacgacg cagaaacacg gccagcgcat ccacaatgtt ttcgccgcta
     1801 tccggatatt cttccagcac ctgaaacgcg gttttgcccg gaatcgcggt ggtcagcagc
     1861 cacgcatcat ccggggtgcg aataaaatgt ttaatggtcg gcagcggcat aaattcggtc
     1921 agccagttca gacgcaccat ttcatcggtc acatcgttcg ccacgctgcc tttgccatgt
     1981 ttcagaaaca gttccggcgc atccggtttg ccatacagac gataaatggt cgcgccgctc
     2041 tgacccacgt tatcacgcgc ccatttatag ccatacagat ccgcatccat gttgctgttc
     2101 agacgcggac ggctacagct cgtttcacgc tgaatatggc tcataacacc ccttgtatta
     2161 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     2221 taacatcaga gattttgaga cacaagatcg gattggcggt tatgcggttg cgatgctggt
     2281 ggctgctgaa cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag
     2341 gtcatcattg acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc
     2401 gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa
     2461 ggtttccagc ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg
     2521 ggccgtcggc gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc
     2581 aaacagcacg acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg
     2641 gtccaggacg cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga
     2701 cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata
     2761 ccggccattg atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg
     2821 ctcgccgata ggggtgcgct tcgcgtactc caagacctgc tgccacacca gttcgtcatc
     2881 gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat
     2941 gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga
     3001 gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa
     3061 ggaaagctgc atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc
     3121 ttcgctgacc tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt
     3181 tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt cgagacgacg
     3241 cgaacgctcc acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc
     3301 gcgctcgatc ttggccgtag cttgctggac tatcgagccg acggactgga aggtttcgcg
     3361 gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc
     3421 gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg
     3481 cgggattgcc ccggaattaa ttccccggat cgatccgtcg atcttgatcc cctgcgccat
     3541 cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca
     3601 gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc
     3661 tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt
     3721 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     3781 ttctacgtgg ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg
     3841 ggggatggga ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt
     3901 cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat
     3961 tggtttaaaa gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt
     4021 gcaaatgctg gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca
     4081 tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc
     4141 gcccctcaag tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg
     4201 gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc
     4261 cggccgaaat cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca
     4321 agtgtcaacg tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca
     4381 acgccggcgg ccctacatgg ctctgctgta gtgagtgggt tgcgctccgg cagcggtcct
     4441 gatcccccgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cccatcatca
//
