LOCUS       sequence-259066-        8916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A binary plasmid containing nopaline synthase promoter to drive 
            bialophos resistance gene (bar) for plant transformation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8916)
  AUTHORS   Testroet A, Lee K, Luth D, Wang K
  TITLE     Comparison of transformation frequency using the bar gene regulated 
            by the CaMV 35S or NOS promoter in Agrobacterium-mediated soybean 
            (Glycine max L.) transformation
  JOURNAL   In Vitro Cell.Dev.Biol.-Plant (2017) 53: 188.
  PUBMED    16249869
REFERENCE   2  (bases 1 to 8916)
  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: "In Vitro 
            Cell.Dev.Biol.-Plant (2017) 53: 188."
COMMENT     Sequence Label: pTF101.4
FEATURES             Location/Qualifiers
     source          1..8916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1217..1846
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2275..3348
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3414..3608
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3844..3866)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3952..4092
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4007..4026
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4107..4124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4278..4866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4358..4377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5112..5903)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6427..6451
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     6651..6673
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6665..6682
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6666..6682
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(7253..7801)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(7863..8042)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    8441..8497
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(8506..8522)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8506..8522)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8519..8541)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8530..8546
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8554..8560,8561..8578,8579..8584))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8599..8620
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    8778..8802
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 agtactttaa agtactttaa agtactttaa agtactttga tccaacccct ccgctgctat
       61 agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag
      121 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt
      181 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag
      241 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc
      301 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc
      361 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac
      421 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt
      481 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac
      541 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag
      601 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg
      661 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc
      721 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc
      781 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt
      841 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc
      901 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg
      961 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct
     1021 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
     1081 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
     1141 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
     1201 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     1261 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
     1321 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
     1381 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
     1441 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
     1501 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
     1561 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
     1621 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
     1681 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
     1741 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
     1801 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
     1861 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
     1921 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
     1981 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
     2041 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat
     2101 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     2161 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     2221 ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
     2281 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     2341 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     2401 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     2461 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     2521 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     2581 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     2641 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     2701 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     2761 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     2821 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     2881 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     2941 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     3001 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     3061 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     3121 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     3181 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     3241 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     3301 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     3361 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat
     3421 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
     3481 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
     3541 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
     3601 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg
     3661 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
     3721 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
     3781 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
     3841 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     3901 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     3961 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     4021 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4081 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4141 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4201 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4261 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4321 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4381 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4441 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4501 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4561 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4621 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4681 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4741 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4801 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4861 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4921 cgttaaggga ttttggtcat gcatgatata tctcccaatt tgtgtagggc ttattatgca
     4981 cgcttaaaaa taataaaagc agacttgacc tgatagtttg gctgtgagca attatgtgct
     5041 tagtgcatct aacgcttgag ttaagccgcg ccgcgaagcg gcgtcggctt gaacgaattt
     5101 ctagctagac attatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat
     5161 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt
     5221 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
     5281 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
     5341 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
     5401 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
     5461 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
     5521 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
     5581 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
     5641 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
     5701 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
     5761 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
     5821 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
     5881 tcggcgatca ccgcttcccc catgatgttt aactttgttt tagggcgact gccctgctgc
     5941 gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc
     6001 ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc catgaaaacc
     6061 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgacggc
     6121 agttacgcta cttgcattac agcttacgaa ccgaacgagg cttatgtcca ctgggttcgt
     6181 gcccgaattg atcacaggca gcaacgctct gtcatcgtta caatcaacat gctaccctcc
     6241 gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga atagcatcgg
     6301 taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt cccggactga
     6361 tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct
     6421 ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac ttaataacac
     6481 attgcggacg tttttaatgt actgaattaa cgccgaattg ctctagcatt cgccattcag
     6541 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6601 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6661 acgttgtaaa acgacggcca gtgccaagct aattcgcttc aagacgtgct caaatcacta
     6721 tttccacacc cctatatttc tattgcactc ccttttaact gttttttatt acaaaaatgc
     6781 cctggaaaat gcactccctt tttgtgtttg tttttttgtg aaacgatgtt gtcaggtaat
     6841 ttatttgtca gtctactatg gtggcccatt atattaatag caactgtcgg tccaatagac
     6901 gacgtcgatt ttctgcattt gtttaaccac gtggatttta tgacatttta tattagttaa
     6961 tttgtaaaac ctacccaatt aaagacctca tatgttctaa agactaatac ttaatgataa
     7021 caattttctt ttagtgaaga aagggataat tagtaaatat ggaacaaggg cagaagattt
     7081 attaaagccg cggtaagaga caacaagtag gtacgtggag tgtcttaggt gacttaccca
     7141 cataacataa agtgacatta acaaacatag ctaatgctcc tatttgaata gtgcatatca
     7201 gcatacctta ttacatatag ataggagcaa actctagcta gattgttgag cagatctcgg
     7261 tgacgggcag gaccggacgg ggcggtaccg gcaggctgaa gtccagctgc cagaaaccca
     7321 cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc
     7381 cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc
     7441 tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc
     7501 ccgtccgctg gtggcggggg gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt
     7561 tgcgtgcctt ccaggggccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga
     7621 cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct
     7681 gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg atggtgcaga
     7741 ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggctca
     7801 tggtagatcc ccaattctcg agcctagagt cgagatctgg attgagagtg aatatgagac
     7861 tctaattgga taccgagggg aatttatgga acgtcagtgg agcatttttg acaagaaata
     7921 tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt ttctgacgta
     7981 tgtgcttagc tcattaaact ccagaaaccc gcggctgagt ggctccttca acgttgcggt
     8041 tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt cataacgtga
     8101 ctcccttaat tctccgctca tgatcttgat cccctgcgcc atcagatcct tggcggcaag
     8161 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     8221 aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca
     8281 ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     8341 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt gttccgcttc
     8401 ctttagcagc ccttgcgccc tgagtgcttg cggcagcgtg aagcttgcat gcctgcaggt
     8461 cgactctaga ggatccccgg gtaccgagct cgaattcgta atcatgtcat agctgtttcc
     8521 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     8581 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     8641 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     8701 gagaggcggt ttgcgtattg gagcttgagc ttggatcaga ttgtcgtttc ccgccttcag
     8761 tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt
     8821 tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta
     8881 tgtgcatgcc aaccacaggg ttcccctcgg gatcaa
//
