LOCUS       sequence-256018-        7030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter plasmid with a 12-nucleotide specific ""barcode"" sequence.
            This plasmid is not available as an individual bacterial stab, but 
            can be requested as part of the Plasmid-ID Kit (Kit #1000000168 )..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7030)
  AUTHORS   Mendoza-Suarez MA, Geddes BA, Sanchez-Canizares C, Ramirez-Gonzalez 
            RH, Kirchhelle C, Jorrin B, Poole PS
  TITLE     Optimizing Rhizobium-legume symbioses by simultaneous measurement of
            rhizobial competitiveness and N2 fixation in nodules.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 May 5;117(18):9822-9831. doi: 
            10.1073/pnas.1921225117. Epub 2020 Apr 21.
  PUBMED    32317381
REFERENCE   2  (bases 1 to 7030)
  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"; doi: 
            "10.1073/pnas.1921225117"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2020-05-5- 5"; volume: "117"; issue: "18"; pages: "9822-9831"
COMMENT     Sequence Label: pOPS0581
FEATURES             Location/Qualifiers
     source          1..7030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      9..103
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(202..221)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             213..1028
                     /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"
     oriT            1179..1287
                     /note="incP origin of transfer"
     CDS             complement(1348..2496)
                     /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      2889..3420
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(5942..6028)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             6275..6988
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(7008..7030)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgctcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
       61 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaggca attgtgtctc
      121 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
      181 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcagcgt gaaacgagct
      241 gtagccgtcc gcgtctgaac agcaacatgg atgcggatct gtatggctat aaatgggcgc
      301 gtgataacgt gggtcagagc ggcgcgacca tttatcgtct gtatggcaaa ccggatgcgc
      361 cggaactgtt tctgaaacat ggcaaaggca gcgtggcgaa cgatgtgacc gatgaaatgg
      421 tgcgtctgaa ctggctgacc gaatttatgc cgctgccgac cattaaacat tttattcgca
      481 ccccggatga tgcgtggctg ctgaccaccg cgattccggg caaaaccgcg tttcaggtgc
      541 tggaagaata tccggatagc ggcgaaaaca ttgtggatgc gctggccgtg tttctgcgtc
      601 gtctgcatag cattccggtg tgcaactgcc cgtttaacag cgatcgtgtg tttcgtctgg
      661 cccaggcgca gagccgtatg aacaacggcc tggtggatgc gagcgatttt gatgatgaac
      721 gtaacggctg gccggtggaa caggtgtgga aagaaatgca taaactgctg ccgtttagcc
      781 cggatagcgt ggtgacccac ggcgatttta gcctggataa cctgattttc gatgaaggca
      841 aactgattgg ctgcattgat gtgggccgtg tgggcattgc ggatcgttat caggatctgg
      901 ccattctgtg gaactgcctg ggcgaattta gcccgagcct gcaaaaacgt ctgtttcaga
      961 aatatggcat tgataatccg gatatgaaca aactgcaatt tcatctgatg ctggatgaat
     1021 ttttctaata attaattgaa ctacttttcc gctgcataac cctgcttcgg ggtcattata
     1081 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     1141 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     1201 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     1261 gggaatcctg ctctgcgagg ctggccgtag gccggccgcg atgcaggtgg ctgctgaacc
     1321 cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac
     1381 ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc
     1441 gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt
     1501 gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga
     1561 cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac
     1621 gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg
     1681 gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat
     1741 cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat
     1801 cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg
     1861 ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag
     1921 ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg
     1981 cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc
     2041 gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat
     2101 ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggctt cgctgacctg
     2161 ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc
     2221 gatggtcatc gacttcgcca aacctgccgc ctcctgttcg agccgacgcg aacgctccac
     2281 ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt
     2341 ggccgtagct tgctggacta tcgagccgac ggactggaag gtttcgcggg gcgcacgcat
     2401 gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc
     2461 ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc
     2521 ggaattaatt ccccggatcg atccgtcgat cttgatcccc tgcgccatca gatccttggc
     2581 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     2641 gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta
     2701 agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc
     2761 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtggct
     2821 gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg
     2881 cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg
     2941 tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc
     3001 aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga
     3061 ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact
     3121 ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg
     3181 tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta
     3241 aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg
     3301 agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc
     3361 cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc
     3421 ctacatggct ctgctgtagt gagtgggttg cgctccggca gcggtcctga tcccccgcag
     3481 aaaaaaagga tctcaagaag atcctttgat cttttctaca ttactgcatg agcttgtgga
     3541 agtgtgctga gcataaatga ttctcctagc tgttctttgg gtaagtacgc catcaggacg
     3601 ttgtgagtgg cgcgattttt agcggctgaa atcagccctt gagcctgtcg gcaagtcgcg
     3661 tcatgaggtc catgcgctca tgcaggatcg ccacgaccaa cgcgggttcg cccgcacgcg
     3721 gcaggcaaaa aacgtagtgg tgttcgcagc gggccatccg cagcgcggga aagagttcgc
     3781 tcatgtcctt aaacgggcct tcgccggcgg caagcctggc tatgccctgt tccagcttag
     3841 cgatatagcg gcgcacctgc gccgcgcccc actcccggcg cgtgtagcgg atgatgccgc
     3901 gtagatcggc ttcggcctca gccgtgagga tgtaggccgt caagcgcgat ccccgctgag
     3961 ttcttcatca agaatttcgc cgacgctctt ggtggacacc ttgccggcaa gcccatcgtt
     4021 gatgcggttc cccagcatgg ttttcagttc ctgccatgcc tgatcggcat cagcgtcacc
     4081 ggggaacaga cgttcgaggg cgtattgctt aatggtcttg ccctgcaagg cggccagggc
     4141 tttcaggctc tggtgctgct ggtccgtcat gtcgattgtc aggcggctca ttggataacc
     4201 tccataaaat acacgtaacc acattagcac atatgtgggc gtgaggctac agcgcgaggc
     4261 gcattaaggt cgggaaaatg cgctaggcgc atttaaattg cgtattgctg taatgcgcca
     4321 tgccggctag actaggccca aatgggtata cccaatttga ccaaggggga cgcgatgagg
     4381 gcggccaagc actaccgaca acttctatcc atcgacttca acatcgaggc gctggccttc
     4441 gtgcctggac ccgacggcac acgcggccgg cgcatccacg tcctggggcg cgaggtccgc
     4501 gaccggcccg gcctggtcga gtacctttcg ccggcgttcg gctcgcgggt ggcgctggac
     4561 ggctactgca aggccaattt cgatgcagtg ctgcacctgg cgtaccccga tcatcagcaa
     4621 tggggccacg catgaagcgc cgaagctacg ccatgctgcg cgccgctgcc gcgctggccg
     4681 tcctggtcgt tgcctcgccg gcatgggccg agctgcgcgg cgaggtcgtg cgcatcatcg
     4741 acggcgacac catcgacgtg ctggtagaca agcagccggt gcgcgtgcgc ctggtggaca
     4801 ttgacgcgcc ggaaaagcgg caagccttcg gcgaacgtgc gcgccaggcg ctggccggca
     4861 tggtgttccg ccggcacgtc ctggtcgacg agaaggacac cgaccgttac ggccgcacgc
     4921 tgggcaccgt ctgggtcaac atggagctgg ccagccggcc gccgcagccg cgcaacgtca
     4981 acgccgcgat ggttcaccag ggcatggcgt gggcctatcg cttccacggc cgcgcggccg
     5041 accctgaaat gctgcggctc gaacaggagg cgcgaggcaa gcgcgtcggc ctctggtccg
     5101 atccgcacgc cgtcgagccg tggaaatggc gacgcgagag caacaaccgg agggacgaag
     5161 gttgaaggtc gcccgcatct acctgcgcgc cagtacggac gagcagaatc ttgaacgcca
     5221 ggagagcctt gtagcggcca cgcgggccgc cgggtactac gtcgccggca tctaccgcga
     5281 gaaggcgtcc ggcgcacgcg ccgaccggcc cgagctgctg cgcatgatcg cggacctgca
     5341 acctggtgaa gtcgtcgttg cggagaagat cgaccgcatc agccgcttgc cgttggccga
     5401 ggccgagcgc ctggttgcgt cgatccgggc caaaggggcc aagctggccg tgcctggcgt
     5461 ggtggacctg tcggagctgg ccgccgaggc gaacggagtg gcgaaaatcg ttctggaatc
     5521 cgtccaggac atgcttttga agctcgcctt gcagatggcc cgcgacgact acgaggatcg
     5581 gcgcgagcgt caacgtcagg gtgtccagtt ggcgaaggcc gccggccgct acaccggccg
     5641 caaacgtgac gccggcatgc acgaccgcat catcacgctt cgctccggcg gatcgagcat
     5701 tgccaagacg gccaagctgg tcggatgcag cccgagccag gtcaaacgag tgtgggcggc
     5761 ctggaacgcg cagcagcaaa aataaagccg ggcagtgccc ggcttttctc accttttcgc
     5821 gtcccgcagg gccgctgcga gcgccctacc tagatcctcg ctttccccct cggtgtagtc
     5881 cggccagggc acgaagggcg cggatgcgaa cctgttgagc aggtcagatc tagggcggcg
     5941 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     6001 ctttcgactg agcctttcgt tttatttgat gcctgccgaa ttcggatccg gagctcgaca
     6061 cgacattgtc gtcacctttg tcgggtccgc gacaagcttt ttgttcctcg atcccgtatg
     6121 ttttcgccta aatatctgaa acagcagcaa attatttttc gttagctcaa tcggcccact
     6181 tggcacgagt cttgaaattt attgcgaggc ggcggagctg ccggccgcat ccgtgcggca
     6241 taaccacggt tgtgaacaaa tgaaggaagg ccaaatgcgt aaaggcgaag agctgttcac
     6301 tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt
     6361 gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac
     6421 tactggtaaa ctgccggtac cttggccgac tctggtaacg acgctgactt atggtgttca
     6481 gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc
     6541 ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg
     6601 tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga
     6661 ctttaaagaa gatggcaata tcctgggcca taagctggaa tacaatttta acagccacaa
     6721 tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca
     6781 caacgttgag gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg
     6841 tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa
     6901 agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat
     6961 cacgcatggt atggatgaac tgtacaaatg agcttatgtc accgctgcgt tttacaacgt
     7021 cgtgactggg
//
