LOCUS       V009976                 8026 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009976
VERSION     V009976
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8026)
  AUTHORS   Collier R, Thomson JG, Thilmony R.
  TITLE     GAANTRY, a versatile and robust Agrobacterium-based gene stacking
            system generates high quality transgenic plants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8026)
  AUTHORS   Thilmony R.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-2017) Crop Improvement and Genetics research Unit,
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   3  (bases 1 to 8026)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8026)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-DEC-2017) Crop Improvement and Genetics research Unit,
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      complement(1..160)
                     /label="CaMV 35S terminator"
                     /note="CaMV 35S terminator"
                     /regulatory_class="terminator"
     CDS             complement(218..1054)
                     /gene="sulI"
                     /label="Dihydropteroate synthase type-1"
                     /note="Dihydropteroate synthase type-1 from Escherichia
                     coli. Accession#: P0C002"
     promoter        complement(1163..1834)
                     /label="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     misc_feature    1959..2013
                     /label="TP901 attB"
                     /note="TP901 attB"
     misc_feature    2035..2059
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        2218..2246
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     CDS             2435..2965
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     misc_feature    3044..3149
                     /label="ParA MRS"
                     /note="ParA MRS"
     primer_bind     complement(3172..3188)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(3196..3212)
                     /label="lac operator"
                     /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(3220..3250)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3265..3286)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3519..3964
                     /label="sacB promoter"
                     /note="sacB promoter and control region"
     CDS             3965..5383
                     /label="SacB"
                     /note="secreted levansucrase that renders bacterial growth
                     sensitive to sucrose"
     rep_origin      complement(5484..6072)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6246..7103)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7104..7208)
                     /label="AmpR promoter"
     rep_origin      complement(7234..7689)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7831..7847
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    7878..7933
                     /label="A118 attB"
                     /note="A118 attB"
ORIGIN
        1 actggatttt ggttttagga attagaaatt ttattgatag aagtatttta caaatacaaa
       61 tacatactaa gggtttctta tatgctcaac acatgagcga aaccctataa gaaccctaat
      121 tcccttatct gggaactact cacacattat tctggagaaa aatagagaga gatagatttg
      181 tagagagaga ctggtgattt ttgcggactc tagactaggc atgatctaac cctcggtctc
      241 tggcgtcgcg actgcgaaat ttcgcgaggg tttccgagat ggtgattgcg cttcgcagat
      301 ctccaggcgc gtgggtgcgg acgtagtcag cgccattgcc gatcgcgtga agttccgccg
      361 caaggctcgc tggacccaga tcctttacag gaaggccaac ggtggcgccc aagaaggatt
      421 tccgcgacac cgagaccaat agcggaagcc ccaacgccga cttcagcttt tgaaggttcg
      481 acagcacgtg cagcgatgtt tccggtgcgg ggctcaagaa aaatcccatc cccggatcga
      541 ggattagccg gtcggcagcg accccgctcc gtcgcaaggc ggaaacccgc gcctcgaaga
      601 accgcacaat ctcgtcgagc gcgtcttcgg gtcgaaggtg accggtgcgg gtggcgatgc
      661 catcccgctg cgctgagtgc ataaccacca gcctgcagtc cgcctcagca atatcgggat
      721 agagcgcagg gtcaggaaat ccttggatat cgttcaggta gcccacgccg cgcttgagcg
      781 catagcgctg ggtttccggt tggaagctgt cgattgaaac acggtgcatc tgatcggaca
      841 gggcgtctaa gagcggcgca atacgtctga tctcatcggc cggcgataca ggcctcgcgt
      901 ccggatggct ggcggccggt ccgacatcca cgacgtctga tccgactcgc agcatttcga
      961 tcgccgcggt gacagcgccg gcggggtcta gccgccggct ctcatcgaag aaggagtcct
     1021 cggtgagatt cagaatgccg aacaccgtca ccatgcactt tactcttcca ccattgcttg
     1081 taatggaagt aatgtcagtg ttgaccttct taactgggaa tccagtcatg gatttgaggc
     1141 cgccgaatgg gcccatctcg aggtcctctc caaatgaaat gaacttcctt atatagagga
     1201 agggtcttgc gaaggatagt gggattgtgc gtcatccctt acgtcagtgg agatatcaca
     1261 tcaatccact tgctttgaag acgtggttgg aacgtcttct ttttccacga tgctcctcgt
     1321 gggtgggggt ccatctttgg gaccactgtc ggcagaggca tcttcaacga tggcctttcc
     1381 tttatcgcaa tgatggcatt tgtaggagcc accttccttt tccactatct tcacaataaa
     1441 gtgacagata gctgggcaat ggaatccgag gaggtttccg gatattaccc tttgttgaaa
     1501 agtctcaatt gccctttggt cttctgagac tgtatctttg atatttttgg agtagacaag
     1561 tgtgtcgtgc tccaccatgt tatcacatca atccacttgc tttgaagacg tggttggaac
     1621 gtcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc
     1681 agaggcatct tcaacgatgg cctttccttt atcgcaatga tggcatttgt aggagccacc
     1741 ttccttttcc actatcttca caataaagtg acagatagct gggcaatgga atccgaggag
     1801 gtttccggat attacccttt gttgaaaagt ctcaattgcc ctttggtctt ctgagactgt
     1861 atctttgata tttttggagt agacaagtgt gtcgtgctcc accatgttga cctgcaggaa
     1921 aggtaccaaa ccatggaaag gcgcgccgtt taaacaaact gataattgcc aacacaatta
     1981 acatctcaat caaggtaaat gctttttgct tttaaagttt aaactatcag tgtttgacag
     2041 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
     2101 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
     2161 ccctcgggag aaacgtacgg acgcacaccg tggaaacgga tgaaggcacg aacccagttg
     2221 acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc acgcaactgg
     2281 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     2341 tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     2401 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcagca acgatgttac
     2461 gcagcagggc agtcgcccta aaacaaagtt aggtggctca agtatgggca tcattcgcac
     2521 atgtaggctc ggccctgacc aagtcaaatc catgcgggct gctcttgatc ttttcggtcg
     2581 tgagttcgga gacgtagcca cctactccca acatcagccg gactccgatt acctcgggaa
     2641 cttgctccgt agtaagacat tcatcgcgct tgctgccttc gaccaagaag cggttgttgg
     2701 cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg cgtagtgaga tctatatcta
     2761 tgatctcgca gtctccggag agcaccggag gcagggcatt gccaccgcgc tcatcaatct
     2821 cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc tacgtgcaag cagattacgg
     2881 tgacgatccc gcagtggctc tctatacaaa gttgggcata cgggaagaag tgatgcactt
     2941 tgatatcgac ccaagtaccg ccacctaaca attcgttcaa gccgagatcg gcttcccggc
     3001 cgcggagttg ttcggtaaat tgtcacaaca ccgccgtacg aaaggtcaaa ttgggtatac
     3061 ccatttgggc ctagtctagc cggcatggcg cattacagca atacgcaatt taaatgcgcc
     3121 tagcgcattt tcccgacctt aatgcgccta aacgcgcttg gcgtaatcat ggtcatagct
     3181 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3241 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3301 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3361 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3421 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3481 atccacagaa tcaggggata acgcaggaaa gaacatggca catatacctg ccgttcacta
     3541 ttatttagtg aaatgagata ttatgatatt ttctgaattg tgattaaaaa ggcaacttta
     3601 tgcccatgca acagaaacta taaaaaatac agagaatgaa aagaaacaga tagatttttt
     3661 agttctttag gcccgtagtc tgcaaatcct tttatgattt tctatcaaac aaaagaggaa
     3721 aatagaccag ttgcaatcca aacgagagtc taatagaatg aggtcgaaaa gtaaatcgcg
     3781 cgggtttgtt actgataaag caggcaagac ctaaaatgtg taaagggcaa agtgtatact
     3841 ttggcgtcac cccttacata ttttaggtct ttttttattg tgcgtaacta acttgccatc
     3901 ttcaaacagg agggctggaa gaagcagacc gctaacacag tacataaaaa aggagacatg
     3961 aacgatgaac atcaaaaagt ttgcaaaaca agcaacagta ttaaccttta ctaccgcact
     4021 gctggcagga ggcgcaactc aagcgtttgc gaaagaaacg aaccaaaagc catataagga
     4081 aacatacggc atttcccata ttacacgcca tgatatgctg caaatccctg aacagcaaaa
     4141 aaatgaaaaa tatcaagttc ctgagttcga ttcgtccaca attaaaaata tctcttctgc
     4201 aaaaggtctg gacgtttggg acagctggcc attacaaaac gctgacggca ctgtcgcaaa
     4261 ctatcacggc taccacatcg tctttgcatt agccggagat cctaaaaatg cggatgacac
     4321 ctcgatttat atgttctatc aaaaagtcgg cgaaacttct attgacagct ggaaaaacgc
     4381 tggccgcgtc tttaaagaca gcgacaaatt cgatgcaaat gattctatcc taaaagacca
     4441 aacacaagaa tggtcaggtt cagccacatt tacatctgac ggaaaaatcc gtttattcta
     4501 cactgatttc tccggtaaac attacggcaa acaaacactg acaactgcac aagtcaacgt
     4561 atcagcatca gacagctctt tgaacatcaa cggtgtagag gactataaat caatctttga
     4621 cggtgacgga aaaacgtatc aaaatgtcca gcagttcatc gacgaaggca actacagctc
     4681 aggcgacaac catacgctga gagatcctca ctacgtagaa gataaaggcc acaaatactt
     4741 agtatttgaa gcaaacactg gaactgaaga tggctaccaa ggcgaagaat ctttatttaa
     4801 caaagcatac tatggcaaaa gcacatcatt cttccgtcaa gaaagtcaaa aacttctgca
     4861 aagcgataaa aaacgcacgg ctgagttagc aaacggcgct ctcggtatga ttgagctaaa
     4921 cgatgattac acactgaaaa aagtgatgaa accgctgatt gcatctaaca cagtaacaga
     4981 tgaaattgaa cgcgcgaacg tctttaaaat gaacggcaaa tggtatctgt tcactgactc
     5041 ccgcggatca aaaatgacga ttgacggcat tacgtctaac gatatttaca tgcttggtta
     5101 tgtttctaat tctttaactg gcccatacaa gccgctgaac aaaactggcc ttgtgttaaa
     5161 aatggatctt gatcctaacg atgtaacctt tacttactca cacttcgctg tacctcaagc
     5221 gaaaggaaac aatgtcgtga ttacaagcta tatgacaaac agaggattct acgcagacaa
     5281 acaatcaacg tttgcgccga gcttcctgct gaacatcaaa ggcaagaaaa catctgttgt
     5341 caaagacagc atccttgaac aaggacaatt aacagtcaac aaataaaaac gcaaaagaaa
     5401 atgccgatat gctatcggca ttacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5461 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5521 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5581 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5641 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5701 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5761 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5821 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5881 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5941 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6001 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6061 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6121 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6181 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6241 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6301 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6361 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6421 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6481 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6541 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6601 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6661 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6721 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6781 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6841 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6901 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6961 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7021 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7081 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7141 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7201 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     7261 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     7321 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     7381 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     7441 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     7501 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     7561 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     7621 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     7681 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     7741 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     7801 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgttaaca
     7861 aaagtacttt tagatctgta actttttcgg atcaagctat gaaggacgca aagagggaac
     7921 taaacactta attacgcgtt ttataacggt cctaaggtag cgaaaaatag ggataacagg
     7981 gtaatcatcc tcgacaaaat cgataaagag ctcaaacctg caggtc
//
