LOCUS       40924_47848        5123 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pYBA-300, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5123)
  AUTHORS   Yan X, Wang H, Ye Y, Zeng G, Ma R, Mi F, Yao L.
  TITLE     pYBA100: An Ease-of-use Binary Vector with LoxP-FRT Recombinase Site
            for Plant Transformation
  JOURNAL   Mol. Plant Breed. 10 (3), 371-379 (2012)
REFERENCE   2  (bases 1 to 5123)
  AUTHORS   Yao L, Yan X, Wang H, Ye Y, Zeng G, Ma R.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-NOV-2012) Beijing Agro-Biotechnology Research Center, 
            Beijing Academy of Agriculture and Forestry Sciences, 9 Shuguang 
            Huayuan Zhonglu, Beijing 100097, P.R. China
REFERENCE   3  (bases 1 to 5123)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5123)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Plant 
            Breed."; date: "2012"; volume: "10"; issue: "3"; pages: "371-379"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-NOV-2012) Beijing Agro-Biotechnology Research Center, Beijing 
            Academy of Agriculture and Forestry Sciences, 9 Shuguang Huayuan 
            Zhonglu, Beijing 100097, P.R. China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      741..1510
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1511..2170
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     misc_feature    2368..2391
                     /label=RB 'overdrive' sequence
                     /note="RB 'overdrive' sequence"
     misc_feature    complement(2392..2416)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    complement(2435..2542)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     protein_bind    2564..2597
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2598..2631
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     terminator      complement(2653..2905)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(2909..3457)
                     /codon_start=1
                     /label=BlpR
                     /note="phosphinothricin acetyltransferase"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(3466..3649)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_recomb     3773..3806
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(3773..3806)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_recomb     3807..3840
                     /label=FRT site
                     /note="FRT site"
     protein_bind    3807..3840
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     misc_feature    complement(3852..3876)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(4066..4857)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag ctagcttgag
      601 tatactcact agactttgct tcgcaaagtc gtgaccgcct acggcggctg cggcgcccta
      661 cgggcttgct ctccgggctt cgccctgcgc ggtcgctgcg ctcccttgcc agcccgtgga
      721 tatgtggacg atggccgcga gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa
      781 ccctgctgga caagctgatg gacaggctgc gcctgcccac gagcttgacc acagggattg
      841 cccaccggct acccagcctt cgaccacata cccaccggct ccaactgcgc ggcctgcggc
      901 cttgccccat caattttttt aattttctct ggggaaaagc ctccggcctg cggcctgcgc
      961 gcttcgcttg ccggttggac accaagtgga aggcgggtca aggctcgcgc agcgaccgcg
     1021 cagcggcttg gccttgacgc gcctggaacg acccaagcct atgcgagtgg gggcagtcga
     1081 aggcgaagcc cgcccgcctg ccccccgagc ctcacggcgg cgagtgcggg ggttccaagg
     1141 gggcagcgcc accttgggca aggccgaagg ccgcgcagtc gatcaacaag ccccggaggg
     1201 gccacttttt gccggagggg gagccgcgcc gaaggcgtgg gggaaccccg caggggtgcc
     1261 cttctttggg caccaaagaa ctagatatag ggcgaaatgc gaaagactta aaaatcaaca
     1321 acttaaaaaa ggggggtacg caacagctca ttgcggcacc ccccgcaata gctcattgcg
     1381 taggttaaag aaaatctgta attgactgcc acttttacgc aacgcataat tgttgtcgcg
     1441 ctgccgaaaa gttgcagctg attgcgcatg gtgccgcaac cgtgcggcac cctaccgcat
     1501 ggagataagc atggccacgc agtccagaga aatcggcatt caagccaaga acaagcccgg
     1561 tcactgggtg caaacggaac gcaaagcgca tgaggcgtgg gccgggctta ttgcgaggaa
     1621 acccacggcg gcaatgctgc tgcatcacct cgtggcgcag atgggccacc agaacgccgt
     1681 ggtggtcagc cagaagacac tttccaagct catcggacgt tctttgcgga cggtccaata
     1741 cgcagtcaag gacttggtgg ccgagcgctg gatctccgtc gtgaagctca acggccccgg
     1801 caccgtgtcg gcctacgtgg tcaatgaccg cgtggcgtgg ggccagcccc gcgaccagtt
     1861 gcgcctgtcg gtgttcagtg ccgccgtggt ggttgatcac gacgaccagg acgaatcgct
     1921 gttggggcat ggcgacctgc gccgcatccc gaccctgtat ccgggcgagc agcaactacc
     1981 gaccggcccc ggcgaggagc cgcccagcca gcccggcatt ccgggcatgg aaccagacct
     2041 gccagccttg accgaaacgg aggaatggga acggcgcggg cagcagcgcc tgccgatgcc
     2101 cgatgagccg tgttttctgg acgatggcga gccgttggag ccgccgacac gggtcacgct
     2161 gccgcgccgg tagcacttgg gttgcgcagc aacccgtaag tgcgctgttc cagactatcg
     2221 gctgtagccg cctcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat
     2281 ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccgt
     2341 ttttatcagg ctctgggagg cagaatacaa acaaacacat acagcgactt agtttacccg
     2401 ccaatatatc ctgtcaacgc gtttaaactc gcgaggtacc gggccccccc tcgaggtcga
     2461 cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta gttctagagc
     2521 ggccgccacc gcggtggagc tccaattggc cctatagacc ggtataactt cgtataatgt
     2581 atgctatacg aagttatgaa gttcctattc tctagaaagt ataggaactt ccgtacgagg
     2641 cctattaatc ccgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg
     2701 cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa
     2761 aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat
     2821 tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac
     2881 tttattgcca aatgtttgaa cgatctcaga tctcggtgac gggcaggacc ggacggggcg
     2941 gaaccggcag gctgaagtcc agctgccaga aacccacgtc atgccagttc ccgtgcttga
     3001 agccggccgc ccgcagcatg ccgcgggggg catatccgag cgcctcgtgc atgcgcacgc
     3061 tcgggtcgtt gggcagcccg atgacagcga ccacgctctt gaagccctgt gcctccaggg
     3121 acttcagcag gtgggtgtag agcgtggagc ccagtcccgt ccgctggtgg cggggggaga
     3181 cgtacacggt agactcggcc gtccagtcgt aggcgttgcg tgccttccag gggccagcgt
     3241 aggcgatgcc ggcgacctcg ccgtccacct cggcgacgag ccagggatag cgctcccgca
     3301 gacggacgag gtcgtccgtc cactcctgcg gttcctgcgg ctcggtacgg aagttgaccg
     3361 tgcttgtctc gatgtagtgg ttgacgatgg tgcagaccgc cggcatgtcc gcctcggtgg
     3421 cacggcggat gtcggccggg cgtcgttctg ggctcataga tccggtgcag attatttgga
     3481 ttgagagtga atatgagact ctaattggat accgagggga atttatggaa cgtcagtgga
     3541 gcatttttga caagaaatat ttgctagctg atagtgacct taggcgactt ttgaacgcgc
     3601 aataatggtt tctgacgtat gtgcttagct cattaaactc cagaaacccg cggctgagtg
     3661 gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc
     3721 ggcgggggtc ataacgtgac tcccttaatt ctccgctcat gatcggcgcg ccataacttc
     3781 gtataatgta tgctatacga agttatgaag ttcctattct ctagaaagta taggaacttc
     3841 acgtgacgcg tgtttacacc acaatatatc ctgccacgat ttgcccatgg gggtgggcga
     3901 agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga
     3961 ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt
     4021 tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag
     4081 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa
     4141 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc
     4201 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt
     4261 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc
     4321 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc
     4381 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg
     4441 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat
     4501 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg
     4561 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac
     4621 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgaag
     4681 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga
     4741 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa
     4801 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg
     4861 aaacgatcct catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg
     4921 cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc
     4981 agctggcaat tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt
     5041 aagcccactg caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc
     5101 ccagtagctg acattcatcc cag
//
