LOCUS       40924_989        8480 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P donor TBS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8480)
  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  JOURNAL   Unpublished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 8480)
  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 8480)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8480)
  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            "Unpublished"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..8480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..2018
                     /note="enhancer blocking insulator; derived from petunia; 
                     transformation booster sequence (TBS)"
                     /regulatory_class="insulator"
     misc_feature    2118..2173
                     /label=A118 attP
                     /note="A118 attP"
     misc_feature    2210..2234
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2566..3357
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     primer_bind     complement(3680..3696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3704..3720)
                     /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(3728..3758)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3773..3794)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4027..4472
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4473..5891
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     rep_origin      complement(5992..6580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6754..7611)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7612..7716)
                     /label=AmpR promoter
     rep_origin      complement(7742..8197)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8339..8355
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8364..8428
                     /label=TP901 attP
                     /note="TP901 attP"
ORIGIN
        1 aagttgtaat gagttgctgg cctctcttgg gattgttcat caaagtagct gtccctacac
       61 accacaacag aatggtgttg tggagagaaa gcacagacac atccttgaga tggcaagggc
      121 acttaagttt cagagtggtg tacctaccag gttttgggga gactgtgtca agactgttgt
      181 atacctgatc aatagactcc ctactccaat cttacaaggc aaatgtccat atgaactact
      241 ctatcagaaa catgccaagc ttgatcactt gagagtgttt gggtgtcttg cttttgcagc
      301 aacactgccc aaaggtgaca aacttgcacc tagagccaag cgaaccatct tcattggtta
      361 ttctgagact caaaagggtt acaggttgta tgacttggat aataaggtga tcatagtgag
      421 cagggatgta gttttcagag agtttcagtt ccctttcaaa gagtccatct cccatgaacc
      481 tgatatgttc actcgagcag gttcacttag ctctgaagat acagctttgc agctagtgta
      541 taatgatatt tttccttcca gctttcatac tgatgatgat gatactccag ttcttattcc
      601 tgacattgta gaggccaatg aggatatcac tttggggact cattctaccc aaaatcatat
      661 cacttcagct gatgctgcac cttcaggcca cctgcatgct cctgcagagt cagttgagtc
      721 agctgagcct gacctagcaa atgctgaatt tgagcaccat actgctgtag ctgatccctc
      781 tctagttcca gcaaacctgc acactagacc taaacgcaat gcaggtcctc ccatctggct
      841 caaggacttt gtgacactga acaaaggctc tagggatgtt ccatacccta tagccaacta
      901 tgtttcctat gatcacttgc ctgttcacta tcaagcttat ttgagtgctt tctccactta
      961 tactgaacct aaatccttca aagaagcagc tcaagatgag aaatggatgg aggccatgtc
     1021 ccttgagata caggctcttg aggataataa cacctgggag attgtccctt taccccctgg
     1081 taaacagcct atagggtcca aatgggtgta caaaattaaa tacaaagcta atggtgaagt
     1141 tgacaggttt aaggcaaggc tagtggccaa gggatacact cagcaagaag gccttgacta
     1201 ccatgaaact ttttctccag tggccaaaat ggtcactgta aaagctgtca tatctgttgc
     1261 tgcttccaag ggctggttcc ttttccagat ggatgtcaac aatgcttttc tacaaggtga
     1321 cctcatggaa gaagtttata tgtctctgcc tcagggtttt cacagccaag gggagaccaa
     1381 agtgtgcagg ctgttgaatc cctctatggt ttaaagcaag catcaaggca gtggaacatc
     1441 aagcttacca ctgtccttat gcaggctggt tttatgcaaa gtgcttatga tcactccttg
     1501 tttaccaaaa gagaggggac tgaccttgtc ataatcctga tttatgtgga tgatctgtta
     1561 ataacaggca gcagcaatgt tttgatttca gaagcaaagg caaccctgca tcagcatttc
     1621 aagatgaaag atttgggaga actaaaatac ttcttaggca ttgaggtgct gagatcagaa
     1681 aagggaatcc tactgaacca gaggaagtat gcactggaac tgatatcagg tgttggtctg
     1741 ggaggctgca gaccagtgtc aaccccaatg gaacagaacc aaaggctgac tactgtagaa
     1801 tatgacaagc acttaggaaa gactgatgat gcagagttag aggatgtggg atcctatcag
     1861 agactggtgg gaaagcttct ctacttgaca atcacaaggc cagatatcag ctttgcagtg
     1921 caggtgctga gtcaattcat gcagcaaccc aaacagtcac acctggaagc agcattgagg
     1981 gttgtgaagt acataaaagg ttctccaggt gttaggaaaa tcacgaattc gtcgacctgc
     2041 agccatggaa aggcgcgcct ttataacggt cctaaggtag cgaaaaatag ggataacagg
     2101 gtaattttac gcgtaaatgt ttagttcctc gttttctctc gttggaagaa gaagaaacga
     2161 gaaactaaaa ttaaaaagta cttttagatc tgtttaaact atcagtgttt gacaggatat
     2221 attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg
     2281 cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttcccctc
     2341 gggagaaacg tacgctcgcg ttggcagcat cacccataat tgtggtttca aaatcggctc
     2401 cgtcgatact atgttatacg ccaactttga aaacaacttt gaaaaagctg ttttctggta
     2461 tttaaagttt tagaatgcaa ggaacagtga attggagttc gtcttgttat aattagcttc
     2521 ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc taaaatgaga
     2581 atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga tacggaagga
     2641 atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata tttaaaaatg
     2701 acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga catgatgcta
     2761 tggctggaag gaaagctgcc tgttccaaag gtcctgcacc ttgaacggca tgatggctgg
     2821 agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta tgaagatgaa
     2881 caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt tcactccatc
     2941 gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga attggattac
     3001 ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga cactccattt
     3061 aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga ggaacttgtc
     3121 ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa agtaagtggc
     3181 tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc cttctgcgtc
     3241 cggtcgatca gggaggatat tggggaagaa cagtatgtcg agctattttt tgacttactg
     3301 gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga attgttttag
     3361 tacctagatg tggcgcaacg atgccggcga caagcaggag cgcaccgact tcttccgcat
     3421 caagtgtttt ggctctcagg ccgaggccca cggcaagtat ttgggcaagg ggtcgctggt
     3481 attcgtgcag ggcaagattc ggaataccaa gtacgagaag gacggccaga cggtctacgg
     3541 gacgtacgaa aggtcaaatt gggtataccc atttgggcct agtctagccg gcatggcgca
     3601 ttacagcaat acgcaattta aatgcgccta gcgcattttc ccgaccttaa tgcgcctaaa
     3661 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3721 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3781 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3841 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3901 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4021 acatggcaca tatacctgcc gttcactatt atttagtgaa atgagatatt atgatatttt
     4081 ctgaattgtg attaaaaagg caactttatg cccatgcaac agaaactata aaaaatacag
     4141 agaatgaaaa gaaacagata gattttttag ttctttaggc ccgtagtctg caaatccttt
     4201 tatgattttc tatcaaacaa aagaggaaaa tagaccagtt gcaatccaaa cgagagtcta
     4261 atagaatgag gtcgaaaagt aaatcgcgcg ggtttgttac tgataaagca ggcaagacct
     4321 aaaatgtgta aagggcaaag tgtatacttt ggcgtcaccc cttacatatt ttaggtcttt
     4381 ttttattgtg cgtaactaac ttgccatctt caaacaggag ggctggaaga agcagaccgc
     4441 taacacagta cataaaaaag gagacatgaa cgatgaacat caaaaagttt gcaaaacaag
     4501 caacagtatt aacctttact accgcactgc tggcaggagg cgcaactcaa gcgtttgcga
     4561 aagaaacgaa ccaaaagcca tataaggaaa catacggcat ttcccatatt acacgccatg
     4621 atatgctgca aatccctgaa cagcaaaaaa atgaaaaata tcaagttcct gagttcgatt
     4681 cgtccacaat taaaaatatc tcttctgcaa aaggtctgga cgtttgggac agctggccat
     4741 tacaaaacgc tgacggcact gtcgcaaact atcacggcta ccacatcgtc tttgcattag
     4801 ccggagatcc taaaaatgcg gatgacacct cgatttatat gttctatcaa aaagtcggcg
     4861 aaacttctat tgacagctgg aaaaacgctg gccgcgtctt taaagacagc gacaaattcg
     4921 atgcaaatga ttctatccta aaagaccaaa cacaagaatg gtcaggttca gccacattta
     4981 catctgacgg aaaaatccgt ttattctaca ctgatttctc cggtaaacat tacggcaaac
     5041 aaacactgac aactgcacaa gtcaacgtat cagcatcaga cagctctttg aacatcaacg
     5101 gtgtagagga ctataaatca atctttgacg gtgacggaaa aacgtatcaa aatgtccagc
     5161 agttcatcga cgaaggcaac tacagctcag gcgacaacca tacgctgaga gatcctcact
     5221 acgtagaaga taaaggccac aaatacttag tatttgaagc aaacactgga actgaagatg
     5281 gctaccaagg cgaagaatct ttatttaaca aagcatacta tggcaaaagc acatcattct
     5341 tccgtcaaga aagtcaaaaa cttctgcaaa gcgataaaaa acgcacggct gagttagcaa
     5401 acggcgctct cggtatgatt gagctaaacg atgattacac actgaaaaaa gtgatgaaac
     5461 cgctgattgc atctaacaca gtaacagatg aaattgaacg cgcgaacgtc tttaaaatga
     5521 acggcaaatg gtatctgttc actgactccc gcggatcaaa aatgacgatt gacggcatta
     5581 cgtctaacga tatttacatg cttggttatg tttctaattc tttaactggc ccatacaagc
     5641 cgctgaacaa aactggcctt gtgttaaaaa tggatcttga tcctaacgat gtaaccttta
     5701 cttactcaca cttcgctgta cctcaagcga aaggaaacaa tgtcgtgatt acaagctata
     5761 tgacaaacag aggattctac gcagacaaac aatcaacgtt tgcgccgagc ttcctgctga
     5821 acatcaaagg caagaaaaca tctgttgtca aagacagcat ccttgaacaa ggacaattaa
     5881 cagtcaacaa ataaaaacgc aaaagaaaat gccgatatgc tatcggcatt acatgtgagc
     5941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6181 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6421 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7741 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     7801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     7861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     7921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     7981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     8041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     8101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     8161 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     8221 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     8281 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     8341 aaaacgacgg ccagtgagcg cgttccaact cgcttaattg cgagttttta tttcgtttat
     8401 ttcaattaag gtaactaaaa aactccttat acctcgagga aacgccggcg aaagcggccc
     8461 aaaaccggtg aattcagatt
//
