LOCUS       40924_944       10366 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY B donor GUS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10366)
  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  JOURNAL   Unpublished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 10366)
  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 10366)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10366)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     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"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(29..281)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(356..2164)
                     /label=GUS
                     /note="beta-glucuronidase"
     CDS             complement(2177..2197)
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     regulatory      complement(2216..4312)
                     /label=Pinus radiata ubiquitin promoter and 5' intron
                     /note="Pinus radiata ubiquitin promoter and 5' intron"
                     /regulatory_class="promoter"
     primer_bind     complement(4327..4343)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4392..4446
                     /label=TP901 attB
                     /note="TP901 attB"
     misc_feature    4468..4492
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        4651..4679
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             4868..5398
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     misc_feature    5477..5582
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(5605..5621)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5629..5645
                     /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(5653..5683)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5698..5719)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5952..6397
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             6398..7816
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     rep_origin      complement(7917..8505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8679..9536)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9537..9641)
                     /label=AmpR promoter
     rep_origin      complement(9667..10122)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     10264..10280
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    10311..10366
                     /label=A118 attB
                     /note="A118 attB"
ORIGIN
        1 acgcgcctta attaagagat ccaattgcga tctagtaaca tagatgacac cgcgcgcgat
       61 aatttatcct agtttgcgcg ctatattttg ttttctatcg cgtattaaat gtataattgc
      121 gggactctaa tcataaaaac ccatctcata aataacgtca tgcattacat gttaattatt
      181 acatgcttaa cgtaattcaa cagaaattat atgataatca tcgcaagacc ggcaacagga
      241 ttcaatctta agaaacttta ttgccaaatg tttgaacgat cggggaaatt cggagctcgg
      301 tagcaattcc cgaggctgta gccgacgatg gtgcgccagg agagttgttg attcattgtt
      361 tgcctccctg ctgcggtttt tcaccgaagt tcatgccagt ccagcgtttt tgcagcagaa
      421 aagccgccga cttcggtttg cggtcgcgag tgaagatccc tttcttgtta ccgccaacgc
      481 gcaatatgcc ttgcgaggtc gcaaaatcgg cgaaattcca tacctgttca ccgacgacgg
      541 cgctgacgcg atcaaagacg cggtgataca tatccagcca tgcacactga tactcttcac
      601 tccacatgtc ggtgtacatt gagtgcagcc cggctaacgt atccacgccg tattcggtga
      661 tgataatcgg ctgatgcagt ttctcctgcc aggccagaag ttctttttcc agtaccttct
      721 ctgccgtttc caaatcgccg ctttggacat accatccgta ataacggttc aggcacagca
      781 catcaaagag atcgctgatg gtatcggtgt gagcgtcgca gaacattaca ttgacgcagg
      841 tgatcggacg cgtcgggtcg agtttacgcg ttgcttccgc cagtggcgcg aaatattccc
      901 gtgcaccttg cggacgggta tccggttcgt tggcaatact ccacatcacc acgcttgggt
      961 ggtttttgtc acgcgctatc agctctttaa tcgcctgtaa gtgcgcttgc tgagtttccc
     1021 cgttgactgc ctcttcgctg tacagttctt tcggcttgtt gcccgcttcg aaaccaatgc
     1081 ctaaagagag gttaaagccg acagcagcag tttcatcaat caccacgatg ccatgttcat
     1141 ctgcccagtc gagcatctct tcagcgtaag ggtaatgcga ggtacggtag gagttggccc
     1201 caatccagtc cattaatgcg tggtcgtgca ccatcagcac gttatcgaat cctttgccac
     1261 gcaagtccgc atcttcatga cgaccaaagc cagtaaagta gaacggtttg tggttaatca
     1321 ggaactgttc gcccttcact gccactgacc ggatgccgac gcgaagcggg tagatatcac
     1381 actctgtctg gcttttggct gtgacgcaca gttcatagag ataaccttca cccggttgcc
     1441 agaggtgcgg attcaccact tgcaaagtcc cgctagtgcc ttgtccagtt gcaaccacct
     1501 gttgatccgc atcacgcagt tcaacgctga catcaccatt ggccaccacc tgccagtcaa
     1561 cagacgcgtg gttacagtct tgcgcgacat gcgtcaccac ggtgatatcg tccacccagg
     1621 tgttcggcgt ggtgtagagc attacgctgc gatggattcc ggcatagtta aagaaatcat
     1681 ggaagtaaga ctgctttttc ttgccgtttt cgtcggtaat caccattccc ggcgggatag
     1741 tctgccagtt cagttcgttg ttcacacaaa cggtgatacg tacacttttc ccggcaataa
     1801 catacggcgt gacatcggct tcaaatggcg tatagccgcc ctgatgctcc atcacttcct
     1861 gattattgac ccacactttg ccgtaatgag tgaccgcatc gaaacgcagc acgatacgct
     1921 ggcctgccca acctttcggt ataaagactt cgcgctgata ccagacgttg cccgcataat
     1981 tacgaatatc tgcatcggcg aactgatcgt taaaactgcc tggcacagca attgcccggc
     2041 tttcttgtaa cgcgctttcc caccaacgct gatcaattcc acagttttcg cgatccagac
     2101 tgaatgccca caggccgtcg agttttttga tttcacgggt tggggtttct acaggacgta
     2161 acatgcgtgg gtcttcaacc tttctcttct tctttggagc catgggcccg gatccttgaa
     2221 atatgactaa cgaatatacc tgcacatcat cacaagctaa ttagacagat acttcaagga
     2281 ataaataatt catcacccaa atacagacca acggacagat tctgcgaaag aagcacaaaa
     2341 gaagagggca gacaccctag caacccatag ttacaacctg aacacaaata ttaacaagaa
     2401 atcaaaaaga aagagacacc ctaaaataca gttgcaatag aaataaaata tggcaccagc
     2461 aatccacgaa ttagaaaaag aaccatcagt tcgaaggata agaggaagaa aatatgtgac
     2521 atgtgacatg tcaatagtaa cgcacaaacc ctaacagctt acacatagag gaccttatag
     2581 ggaaagcaca tcactggaat aatcacctac cctcagaacc cgggcccaaa attaaaatat
     2641 tactacagaa cgccacaaga aaatgctcaa agaagaaagg gttcctgtga gatcaaagga
     2701 gggaaactat aacaaggttg gagacaaaat attaaagtac aacgcaataa tctgcaaagg
     2761 ccttctgaaa cacatgacgc aataaaacgc aaacacctgc tgaaaccttt acagacaaga
     2821 gcctattaca aacagcgact actcataact tacatacgaa aattaaatat tccagagaga
     2881 cagacctatt aagggaagaa aacacagatg aaagcacgaa agccctagta atgtcagcca
     2941 tagtcgagaa aaataaccca aataaacatg cgaaacccaa aaggacctgg caaaagattc
     3001 ttcccccaca gaaaccacca ttaacaattt acaaagaaaa acaaatgtta agagtaaagc
     3061 ccttcaaaac tccatacctt tgaatgaaaa gctctctgag aatatcggaa ttcgatgggt
     3121 tgaatgaaaa gctctctgag aatatcgcac tgattataca acgcaagaga aaagatgaga
     3181 agtaacgcgg tgaaggcgag tatttaatag cgttggatga gacccttctc ccgctacgcg
     3241 acacgacaac agacgacctg tcaccatcgg tggttgcgta ccagcgggga taacaacgac
     3301 attttccaat agagaaatga cacgtgtata ctgtgcgctg gctgtgtcgg tgggttatac
     3361 agggtcatcc ggtcgcacat ccgtcacgct tctcgagacg tatgtaaagc gcgtctgtag
     3421 gattccaact ccgcgtggac cccacaataa attacttttt aatacaaaac cagggagata
     3481 aaagattctc gttcgaggga tctccgatat gtcattatgc catttaagta aaataaataa
     3541 actttttgtt tttttaagta aataaaagaa ataaattgtt ggtttgtgaa tacttttatt
     3601 tactttattt ctttatctac tcttaaggaa tgtaattaac ggtgaagtag gaaaagtttt
     3661 acggttaggg gaggagttgc tttgaaagtt atgaaaattc tagttatatt tagagaaaat
     3721 ccgatacgga tttttgtaga tccaagttat ttttgttaaa cttcaaaatt ttaggtgtcc
     3781 tgaatgtttt tgatggtgct gaatctattt ccatcataaa aaacatgatt caagcactcc
     3841 ctctacaaaa aaattttaat cttttttacc gggattccgg ccacttttaa ctgtctaggg
     3901 aattcctccg cctcctggtt agacgattcc aatgtcttat cctctcaaac ttgtacatca
     3961 atatttgttc actttggttt tctttttgac ttaataatac atagattttt tactctataa
     4021 tccttggatt ttcttgaaag taagtttggt agatggattt ttccattgat aaatcttgtt
     4081 gacttggtga acatctttcc aactatgggc aatttgtcca agtggatctt ctagactcct
     4141 ttgagtggat atatttatac aattttgata tgatttagga agtaatgtag tttttttata
     4201 tcaatgtagt tagttttttc caacattcag gattgtaacg gttggtggga aaggtagtgg
     4261 agagtttgat aattaaataa agaatttttt ttatgtattt gtgaggggtt ttcccatcaa
     4321 gcttatcgat accgtcgacc tcgagggggg gcccggtacc aaaccatgga aaggcgcgcc
     4381 gtttaaacaa actgataatt gccaacacaa ttaacatctc aatcaaggta aatgcttttt
     4441 gcttttaaag tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga
     4501 aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg
     4561 tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gagaaacgta cggacgcaca
     4621 ccgtggaaac ggatgaaggc acgaacccag ttgacataag cctgttcggt tcgtaaactg
     4681 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
     4741 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt acagtctatg
     4801 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     4861 agcaacgatg ttacgcagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     4921 gttaggtggc tcaagtatgg gcatcattcg cacatgtagg ctcggccctg accaagtcaa
     4981 atccatgcgg gctgctcttg atcttttcgg tcgtgagttc ggagacgtag ccacctactc
     5041 ccaacatcag ccggactccg attacctcgg gaacttgctc cgtagtaaga cattcatcgc
     5101 gcttgctgcc ttcgaccaag aagcggttgt tggcgctctc gcggcttacg ttctgcccaa
     5161 gtttgagcag ccgcgtagtg agatctatat ctatgatctc gcagtctccg gagagcaccg
     5221 gaggcagggc attgccaccg cgctcatcaa tctcctcaag catgaggcca acgcgcttgg
     5281 tgcttatgtg atctacgtgc aagcagatta cggtgacgat cccgcagtgg ctctctatac
     5341 aaagttgggc atacgggaag aagtgatgca ctttgatatc gacccaagta ccgccaccta
     5401 acaattcgtt caagccgaga tcggcttccc ggccgcggag ttgttcggta aattgtcaca
     5461 acaccgccgt acgaaaggtc aaattgggta tacccatttg ggcctagtct agccggcatg
     5521 gcgcattaca gcaatacgca atttaaatgc gcctagcgca ttttcccgac cttaatgcgc
     5581 ctaaacgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5641 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5701 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5761 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5821 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5881 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5941 aaagaacatg gcacatatac ctgccgttca ctattattta gtgaaatgag atattatgat
     6001 attttctgaa ttgtgattaa aaaggcaact ttatgcccat gcaacagaaa ctataaaaaa
     6061 tacagagaat gaaaagaaac agatagattt tttagttctt taggcccgta gtctgcaaat
     6121 ccttttatga ttttctatca aacaaaagag gaaaatagac cagttgcaat ccaaacgaga
     6181 gtctaataga atgaggtcga aaagtaaatc gcgcgggttt gttactgata aagcaggcaa
     6241 gacctaaaat gtgtaaaggg caaagtgtat actttggcgt caccccttac atattttagg
     6301 tcttttttta ttgtgcgtaa ctaacttgcc atcttcaaac aggagggctg gaagaagcag
     6361 accgctaaca cagtacataa aaaaggagac atgaacgatg aacatcaaaa agtttgcaaa
     6421 acaagcaaca gtattaacct ttactaccgc actgctggca ggaggcgcaa ctcaagcgtt
     6481 tgcgaaagaa acgaaccaaa agccatataa ggaaacatac ggcatttccc atattacacg
     6541 ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa aaatatcaag ttcctgagtt
     6601 cgattcgtcc acaattaaaa atatctcttc tgcaaaaggt ctggacgttt gggacagctg
     6661 gccattacaa aacgctgacg gcactgtcgc aaactatcac ggctaccaca tcgtctttgc
     6721 attagccgga gatcctaaaa atgcggatga cacctcgatt tatatgttct atcaaaaagt
     6781 cggcgaaact tctattgaca gctggaaaaa cgctggccgc gtctttaaag acagcgacaa
     6841 attcgatgca aatgattcta tcctaaaaga ccaaacacaa gaatggtcag gttcagccac
     6901 atttacatct gacggaaaaa tccgtttatt ctacactgat ttctccggta aacattacgg
     6961 caaacaaaca ctgacaactg cacaagtcaa cgtatcagca tcagacagct ctttgaacat
     7021 caacggtgta gaggactata aatcaatctt tgacggtgac ggaaaaacgt atcaaaatgt
     7081 ccagcagttc atcgacgaag gcaactacag ctcaggcgac aaccatacgc tgagagatcc
     7141 tcactacgta gaagataaag gccacaaata cttagtattt gaagcaaaca ctggaactga
     7201 agatggctac caaggcgaag aatctttatt taacaaagca tactatggca aaagcacatc
     7261 attcttccgt caagaaagtc aaaaacttct gcaaagcgat aaaaaacgca cggctgagtt
     7321 agcaaacggc gctctcggta tgattgagct aaacgatgat tacacactga aaaaagtgat
     7381 gaaaccgctg attgcatcta acacagtaac agatgaaatt gaacgcgcga acgtctttaa
     7441 aatgaacggc aaatggtatc tgttcactga ctcccgcgga tcaaaaatga cgattgacgg
     7501 cattacgtct aacgatattt acatgcttgg ttatgtttct aattctttaa ctggcccata
     7561 caagccgctg aacaaaactg gccttgtgtt aaaaatggat cttgatccta acgatgtaac
     7621 ctttacttac tcacacttcg ctgtacctca agcgaaagga aacaatgtcg tgattacaag
     7681 ctatatgaca aacagaggat tctacgcaga caaacaatca acgtttgcgc cgagcttcct
     7741 gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac agcatccttg aacaaggaca
     7801 attaacagtc aacaaataaa aacgcaaaag aaaatgccga tatgctatcg gcattacatg
     7861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8341 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8521 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8581 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8641 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8701 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8761 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8821 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8881 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8941 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     9001 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     9061 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     9121 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     9181 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     9241 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     9301 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     9361 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     9421 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9481 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9541 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9601 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9661 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     9721 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     9781 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     9841 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     9901 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     9961 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
    10021 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
    10081 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
    10141 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
    10201 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
    10261 gttgtaaaac gacggccagt gagcgcgtta acaaaagtac ttttagatct gtaacttttt
    10321 cggatcaagc tatgaaggac gcaaagaggg aactaaacac ttaatt
//
