LOCUS       sequence-263755-        8912 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 8912)
  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 8912)
  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..8912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1337..1966
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2395..3468
                     /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      3534..3728
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3964..3986)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4072..4212
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4127..4146
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4227..4244)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4398..4986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4478..4497)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5232..6023)
                     /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    6544..6568
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     6768..6790
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6782..6799
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6783..6799
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(7369..7917)
                     /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(7979..8158)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    8557..8613
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(8622..8638)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8622..8638)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8635..8657)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8646..8662
                     /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(8670..8676,8677..8694,8695..8700))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8715..8736
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    join(8894..8912,1..6)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa
       61 ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc tcgggatcaa
      121 agtactttaa agtactttaa agtactttaa agtactttga tccaacccct ccgctgctat
      181 agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag
      241 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt
      301 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag
      361 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc
      421 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc
      481 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac
      541 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt
      601 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac
      661 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag
      721 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg
      781 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc
      841 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc
      901 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt
      961 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc
     1021 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg
     1081 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct
     1141 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
     1201 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
     1261 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
     1321 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
     1381 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
     1441 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
     1501 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
     1561 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
     1621 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
     1681 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
     1741 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
     1801 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
     1861 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
     1921 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
     1981 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
     2041 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
     2101 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
     2161 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat
     2221 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
     2281 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
     2341 ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
     2401 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
     2461 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
     2521 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
     2581 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
     2641 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
     2701 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
     2761 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
     2821 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
     2881 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
     2941 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
     3001 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
     3061 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
     3121 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
     3181 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
     3241 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
     3301 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
     3361 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
     3421 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
     3481 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat
     3541 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
     3601 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
     3661 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
     3721 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg
     3781 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
     3841 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
     3901 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
     3961 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4021 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     4081 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     4141 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     4201 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4261 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4321 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4381 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4441 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4501 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4561 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4621 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4681 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4741 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4801 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4861 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4921 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4981 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5041 cgttaaggga ttttggtcat gcatgatata tctcccaatt tgtgtagggc ttattatgca
     5101 cgcttaaaaa taataaaagc agacttgacc tgatagtttg gctgtgagca attatgtgct
     5161 tagtgcatct aacgcttgag ttaagccgcg ccgcgaagcg gcgtcggctt gaacgaattt
     5221 ctagctagac attatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat
     5281 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt
     5341 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
     5401 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
     5461 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
     5521 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
     5581 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
     5641 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
     5701 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
     5761 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
     5821 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
     5881 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
     5941 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
     6001 tcggcgatca ccgcttcccc catgatgttt aactttgttt tagggcgact gccctgctgc
     6061 gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc
     6121 ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc catgaaaacc
     6181 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc
     6241 atacgctact tgcattacag cttacgaacc gaacaggctt atgtccactg ggttcgtgcc
     6301 cgaattgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct accctccgcg
     6361 agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata gcatcggtaa
     6421 catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc ggactgatgg
     6481 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc
     6541 tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt
     6601 gcggacgttt ttaatgtact gaattaacgc cgaattgctc tagcattcgc cattcaggct
     6661 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6721 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6781 ttgtaaaacg acggccagtg ccaagctaat tcgcttcaag acgtgctcaa atcactattt
     6841 ccacacccct atatttctat tgcactccct tttaactgtt ttttattaca aaaatgccct
     6901 ggaaaatgca ctcccttttt gtgtttgttt ttttgtgaaa cgatgttgtc aggtaattta
     6961 tttgtcagtc tactatggtg gcccattata ttaatagcaa ctgtcggtcc aatagacgac
     7021 gtcgattttc tgcatttgtt taaccacgtg gattttatga cattttatat tagttaattt
     7081 gtaaaaccta cccaattaaa gacctcatat gttctaaaga ctaatactta atgataacaa
     7141 ttttctttta gtgaagaaag ggataattag taaatatgga acaagggcag aagatttatt
     7201 aaagccgcgt aagagacaac aagtaggtac gtggagtgtc ttaggtgact tacccacata
     7261 acataaagtg acattaacaa acatagctaa tgctcctatt tgaatagtgc atatcagcat
     7321 accttattac atatagatag gagcaaactc tagctagatt gttgagcaga tctcggtgac
     7381 gggcaggacc ggacggggcg gtaccggcag gctgaagtcc agctgccaga aacccacgtc
     7441 atgccagttc ccgtgcttga agccggccgc ccgcagcatg ccgcgggggg catatccgag
     7501 cgcctcgtgc atgcgcacgc tcgggtcgtt gggcagcccg atgacagcga ccacgctctt
     7561 gaagccctgt gcctccaggg acttcagcag gtgggtgtag agcgtggagc ccagtcccgt
     7621 ccgctggtgg cggggggaga cgtacacggt cgactcggcc gtccagtcgt aggcgttgcg
     7681 tgccttccag gggcccgcgt aggcgatgcc ggcgacctcg ccgtccacct cggcgacgag
     7741 ccagggatag cgctcccgca gacggacgag gtcgtccgtc cactcctgcg gttcctgcgg
     7801 ctcggtacgg aagttgaccg tgcttgtctc gatgtagtgg ttgacgatgg tgcagaccgc
     7861 cggcatgtcc gcctcggtgg cacggcggat gtcggccggg cgtcgttctg ggctcatggt
     7921 agatccccaa ttctcgagcc tagagtcgag atctggattg agagtgaata tgagactcta
     7981 attggatacc gaggggaatt tatggaacgt cagtggagca tttttgacaa gaaatatttg
     8041 ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct gacgtatgtg
     8101 cttagctcat taaactccag aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg
     8161 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
     8221 cttaattctc cgctcatgat cttgatcccc tgcgccatca gatccttggc ggcaagaaag
     8281 ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt
     8341 ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc
     8401 aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga
     8461 cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtgttc cgcttccttt
     8521 agcagccctt gcgccctgag tgcttgcggc agcgtgaagc ttgcatgcct gcaggtcgac
     8581 tctagaggat ccccgggtac cgagctcgaa ttcgtaatca tgtcatagct gtttcctgtg
     8641 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     8701 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     8761 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     8821 ggcggtttgc gtattggagc ttgagcttgg atcagattgt cgtttcccgc cttcagttta
     8881 aactatcagt gtttgacagg atatattggc gg
//
