LOCUS       sequence-296926-        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    133..157
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    184..208
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        250..284
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             293..304
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      993..1064
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1080..1107
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1338..2153)
                     /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     2145..2164
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(2329..3477)
                     /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      3870..4401
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ccatcatcag ttcggtggtc ttccgacgaa caataaggcc gcaaatcgcg gcctttttta
       61 ttgataacaa aaccggctca gttctgcgta gaaaccaaca tgcaagctcc accgggtgca
      121 aagcggcagc ggcggcagga tatattcaat tgtaaatggc ttcatgtccg ggaaatctac
      181 tggtggcagg atatattgtg gtgtaaacaa tggagaaaaa gagaattcac ctgcatatgg
      241 agagagacct ttacggctag ctcagtccta ggtatagtgc tagctactag agaaagagga
      301 gaaatactaa atgtcgcact caaaacaagc actggcagat acgatgaaaa tgacgtggct
      361 gatggaaggt tcggtcaacg gtcacgcctt tacgattgaa ggcgaaggta ccggcaaacc
      421 gtatgaaggt aaacagagtg gcacctttcg tgttacgaaa ggcggtccgc tgccgtttgc
      481 gtttgatatt gtcgctccga ccctgaaata cggtttcaaa tgcttcatga aatacccggc
      541 ggatattccg gactacttta aactggcctt cccggaaggc ctgacgtacg atcgtaaaat
      601 tgcgtttgaa gacggcggtt gtgcgaccgc cacggtcgaa atgagcctga agggtaacac
      661 cctggtgcat aaaacgaact ttcagggcgg taatttcccg attgatggtc cggtgatgca
      721 aaaacgcacc ctgggctggg aaccgacctc tgaaaaaatg acgccgtgcg atggtattat
      781 caaaggcgac accatcatgt acctgatggt tgaaggcggt aaaacgctga aatgtcgtta
      841 tgaaaacaat taccgcgcca acaaaccagt gctgatgccg ccgagccact ttgtggatct
      901 gcgtctgacc cgcacgaatc tggataaaga aggtctggcg ttcaaactgg aagaatatgc
      961 tgttgcccgt gtgctggaag tgtaataagc ttcaaataaa acgaaaggct cagtcgaaag
     1021 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     1081 gctcaccttc gggtgggcct ttctgcgggt ctcacgctca gaatatgcag gtgcctgcag
     1141 gaattgttga ttttgtgatg actgatggca ggatatatgc ggttgtaatt catttttatt
     1201 gtctaaattt ctgtatttgt ttgtttgttc ggttgtaaat ttttttggaa gaacaagaaa
     1261 agaaaaaaca cccgttaggg tgtttttagt tagtgtggcg cgccgacttg cgacatgcgg
     1321 tcctttgcaa tcaactatta gaaaaattca tccagcatca gatgaaattg cagtttgttc
     1381 atatccggat tatcaatgcc atatttctga aacagacgtt tttgcaggct cgggctaaat
     1441 tcgcccaggc agttccacag aatggccaga tcctgataac gatccgcaat gcccacacgg
     1501 cccacatcaa tgcagccaat cagtttgcct tcatcgaaaa tcaggttatc caggctaaaa
     1561 tcgccgtggg tcaccacgct atccgggcta aacggcagca gtttatgcat ttctttccac
     1621 acctgttcca ccggccagcc gttacgttca tcatcaaaat cgctcgcatc caccaggccg
     1681 ttgttcatac ggctctgcgc ctgggccaga cgaaacacac gatcgctgtt aaacgggcag
     1741 ttgcacaccg gaatgctatg cagacgacgc agaaacacgg ccagcgcatc cacaatgttt
     1801 tcgccgctat ccggatattc ttccagcacc tgaaacgcgg ttttgcccgg aatcgcggtg
     1861 gtcagcagcc acgcatcatc cggggtgcga ataaaatgtt taatggtcgg cagcggcata
     1921 aattcggtca gccagttcag acgcaccatt tcatcggtca catcgttcgc cacgctgcct
     1981 ttgccatgtt tcagaaacag ttccggcgca tccggtttgc catacagacg ataaatggtc
     2041 gcgccgctct gacccacgtt atcacgcgcc catttatagc catacagatc cgcatccatg
     2101 ttgctgttca gacgcggacg gctacagctc gtttcacgct gaatatggct cataacaccc
     2161 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     2221 tgtgcaatgt aacatcagag attttgagac acaagatcgg attggcggtt atgcggttgc
     2281 gatgctggtg gctgctgaac ccccagccgg aactgacccc acaaggccct agcgtttgca
     2341 atgcaccagg tcatcattga cccaggcgtg ttccaccagg ccgctgcctc gcaactcttc
     2401 gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg gtggaatccg atccgcacat
     2461 gaggcggaag gtttccagct tgagcgggta cggctcccgg tgcgagctga aatagtcgaa
     2521 catccgtcgg gccgtcggcg acagcttgcg gtacttctcc catatgaatt tcgtgtagtg
     2581 gtcgccagca aacagcacga cgatttcctc gtcgatcagg acctggcaac gggacgtttt
     2641 cttgccacgg tccaggacgc ggaagcggtg cagcagcgac accgattcca ggtgcccaac
     2701 gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt cctcggcctt
     2761 cgtgtaatac cggccattga tcgaccagcc caggtcctgg caaagctcgt agaacgtgaa
     2821 ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc aagacctgct gccacaccag
     2881 ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt ccttgttgac
     2941 gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc gcgtggtgaa
     3001 cagggcagag cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc acggcgcaat
     3061 atcgaacaag gaaagctgca tttccttgat ctgctgcttc gtgtgtttca gcaacgcggc
     3121 ctgcttggct tcgctgacct gttttgccag gtcctcgccg gcggtttttc gcttcttggt
     3181 cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg cctcctgttc
     3241 gagacgacgc gaacgctcca cggcggccga tggcgcgggc agggcagggg gagccagttg
     3301 cacgctgtcg cgctcgatct tggccgtagc ttgctggact atcgagccga cggactggaa
     3361 ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat cctcggcgga
     3421 aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc gattcattca
     3481 ccctccttgc gggattgccc cggaattaat tccccggatc gatccgtcga tcttgatccc
     3541 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     3601 accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc
     3661 cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg
     3721 ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc
     3781 ggactggctt tctacgtggc tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc
     3841 agcccctggg gggatgggag gcccgcgtta gcgggccggg agggttcgag aagggggggc
     3901 accccccttc ggcgtgcgcg gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt
     3961 tataaatatt ggtttaaaag caggttaaaa gacaggttag cggtggccga aaaacgggcg
     4021 gaaacccttg caaatgctgg attttctgcc tgtggacagc ccctcaaatg tcaataggtg
     4081 cgcccctcat ctgtcagcac tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc
     4141 agtagtcgcg cccctcaagt gtcaataccg cagggcactt atccccaggc ttgtccacat
     4201 catctgtggg aaactcgcgt aaaatcaggc gttttcgccg atttgcgagg ctggccagct
     4261 ccacgtcgcc ggccgaaatc gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc
     4321 ggcccctcaa gtgtcaacgt ccgcccctca tctgtcagtg agggccaagt tttccgcgag
     4381 gtatccacaa cgccggcggc cctacatggc tctgctgtag tgagtgggtt gcgctccggc
     4441 agcggtcctg atcccccgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
//
