LOCUS       40924_984        8526 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P donor nptII, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8526)
  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  JOURNAL   Unpublished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 8526)
  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 8526)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8526)
  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            "Unpublished"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..8526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..811
                     /label=Arabidopsis ubiquitin 10 promoter and 5' intron
                     /note="Arabidopsis ubiquitin 10 promoter and 5' intron"
                     /regulatory_class="promoter"
     CDS             827..1615
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      1679..1931
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2086..2141
                     /label=A118 attP
                     /note="A118 attP"
     misc_feature    2178..2202
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2534..3325
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    3520..3625
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(3648..3664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3672..3688)
                     /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(3696..3726)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3741..3762)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3995..4440
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4441..5859
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     rep_origin      complement(5960..6548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6722..7579)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7580..7684)
                     /label=AmpR promoter
     rep_origin      complement(7710..8165)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8307..8323
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8332..8396
                     /label=TP901 attP
                     /note="TP901 attP"
ORIGIN
        1 ttctagaaaa tacgaaatac gcttcaatgc agtgggaccc acggttcaat tattgccaat
       61 tttcagctcc accgtatatt taaaaaataa aacgataatg ctaaaaaaat ataaatcgta
      121 acgatcgtta aatctcaacg gctggatctt atgacgaccg ttagaaattg tggttgtcga
      181 cgagtcagta ataaacggcg tcaaagtggt tgcagccggc acacacgagt cgtgtttatc
      241 aactcaaagc acaaatactt ttcctcaacc taaaaataag gcaattagcc aaaaacaact
      301 ttgcgtgtaa acaacgctca atacacgtgt cattttatta ttagctattg cttcaccgcc
      361 ttagctttct cgtgacctag tcgtcctcgt cttttcttct tcttcttcta taaaacaata
      421 cccaaagagc tcttcttctt cacaattcag atttcaattt ctcaaaatct taaaaacttt
      481 ctctcaattc tctctaccgt gatcaaggta aatttctgtg ttccttattc tctcaaaatc
      541 ttcgattttg ttttcgttcg atcccaattt cgtatatgtt ctttggttta gattctgtta
      601 atcttagatc gaagacgatt ttctgggttt gatcgttaga tatcatctta attctcgatt
      661 agggtttcat agatatcatc cgatttgttc aaataatttg agttttgtcg aataattact
      721 cttcgatttg tgatttctat ctagatctgg tgttagtttc tagtttgtgc gatcgaattt
      781 gtcgattaat ctgagttttt ctgattaaca gggcgcgtcc atggggattg aacaagatgg
      841 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
      901 acagacaatc ggctgctctg atgccgccgt gtttcggctg tcagcgcagg ggcgcccggt
      961 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctccaggacg aggcagcgcg
     1021 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     1081 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     1141 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     1201 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     1261 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     1321 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     1381 gacacatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     1441 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     1501 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     1561 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaac
     1621 tagtggtacc gccccgtccg gtcctgcccg tcaccgagat ctgatccgtc gacctgcaga
     1681 tcgttcaaac atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat
     1741 gattatcata taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat
     1801 gacgttattt atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc
     1861 gatagaaaac aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat
     1921 gttactagat ccgatgataa gctgtcaaac atgagaattc aaagtcgaca aaatcgataa
     1981 agagctcaaa cctgcaggaa aggtaccaaa ccatggaaag gcgcgccttt ataacggtcc
     2041 taaggtagcg aaaaataggg ataacagggt aattttacgc gtaaatgttt agttcctcgt
     2101 tttctctcgt tggaagaaga agaaacgaga aactaaaatt aaaaagtact tttagatctg
     2161 tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt
     2221 tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta
     2281 tgtgcatgcc aaccacaggg ttcccctcgg gagaaacgta cgctcgcgtt ggcagcatca
     2341 cccataattg tggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa
     2401 acaactttga aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat
     2461 tggagttcgt cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg
     2521 aaggaaataa taaatggcta aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa
     2581 ataccgctgc gtaaaagata cggaaggaat gtctcctgct aaggtatata agctggtggg
     2641 agaaaatgaa aacctatatt taaaaatgac ggacagccgg tataaaggga ccacctatga
     2701 tgtggaacgg gaaaaggaca tgatgctatg gctggaagga aagctgcctg ttccaaaggt
     2761 cctgcacttt gaacggcatg atggctggag caatctgctc atgagtgagg ccgatggcgt
     2821 cctttgctcg gaagagtatg aagatgaaca aagccctgaa aagattatcg agctgtatgc
     2881 ggagtgcatc aggctctttc actccatcga catatcggat tgtccctata cgaatagctt
     2941 agacagccgc ttagccgaat tggattactt actgaataac gatctggccg atgtggattg
     3001 cgaaaactgg gaagaagaca ctccatttaa agatccgcgc gagctgtatg attttttaaa
     3061 gacggaaaag cccgaagagg aacttgtctt ttcccacggc gacctgggag acagcaacat
     3121 ctttgtgaaa gatggcaaag taagtggctt tattgatctt gggagaagcg gcagggcgga
     3181 caagtggtat gacattgcct tctgcgtccg gtcgatcagg gaggatattg gggaagaaca
     3241 gtatgtcgag ctattttttg acttactggg gatcaagcct gattgggaga aaataaaata
     3301 ttatatttta ctggatgaat tgttttagta cctagatgtg gcgcaacgat gccggcgaca
     3361 agcaggagcg caccgacttc ttccgcatca agtgttttgg ctctcaggcc gaggcccacg
     3421 gcaagtattt gggcaagggg tcgctggtat tcgtgcaggg caagattcgg aataccaagt
     3481 acgagaagga cggccagacg gtctacggga cgtacgaaag gtcaaattgg gtatacccat
     3541 ttgggcctag tctagccggc atggcgcatt acagcaatac gcaatttaaa tgcgcctagc
     3601 gcattttccc gaccttaatg cgcctaaacg cgcttggcgt aatcatggtc atagctgttt
     3661 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3721 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3841 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3901 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3961 acagaatcag gggataacgc aggaaagaac atggcacata tacctgccgt tcactattat
     4021 ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca actttatgcc
     4081 catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga ttttttagtt
     4141 ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata
     4201 gaccagttgc aatccaaacg agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg
     4261 tttgttactg ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg tatactttgg
     4321 cgtcacccct tacatatttt aggtcttttt ttattgtgcg taactaactt gccatcttca
     4381 aacaggaggg ctggaagaag cagaccgcta acacagtaca taaaaaagga gacatgaacg
     4441 atgaacatca aaaagtttgc aaaacaagca acagtattaa cctttactac cgcactgctg
     4501 gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca
     4561 tacggcattt cccatattac acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat
     4621 gaaaaatatc aagttcctga gttcgattcg tccacaatta aaaatatctc ttctgcaaaa
     4681 ggtctggacg tttgggacag ctggccatta caaaacgctg acggcactgt cgcaaactat
     4741 cacggctacc acatcgtctt tgcattagcc ggagatccta aaaatgcgga tgacacctcg
     4801 atttatatgt tctatcaaaa agtcggcgaa acttctattg acagctggaa aaacgctggc
     4861 cgcgtcttta aagacagcga caaattcgat gcaaatgatt ctatcctaaa agaccaaaca
     4921 caagaatggt caggttcagc cacatttaca tctgacggaa aaatccgttt attctacact
     4981 gatttctccg gtaaacatta cggcaaacaa acactgacaa ctgcacaagt caacgtatca
     5041 gcatcagaca gctctttgaa catcaacggt gtagaggact ataaatcaat ctttgacggt
     5101 gacggaaaaa cgtatcaaaa tgtccagcag ttcatcgacg aaggcaacta cagctcaggc
     5161 gacaaccata cgctgagaga tcctcactac gtagaagata aaggccacaa atacttagta
     5221 tttgaagcaa acactggaac tgaagatggc taccaaggcg aagaatcttt atttaacaaa
     5281 gcatactatg gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc
     5341 gataaaaaac gcacggctga gttagcaaac ggcgctctcg gtatgattga gctaaacgat
     5401 gattacacac tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt aacagatgaa
     5461 attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt atctgttcac tgactcccgc
     5521 ggatcaaaaa tgacgattga cggcattacg tctaacgata tttacatgct tggttatgtt
     5581 tctaattctt taactggccc atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg
     5641 gatcttgatc ctaacgatgt aacctttact tactcacact tcgctgtacc tcaagcgaaa
     5701 ggaaacaatg tcgtgattac aagctatatg acaaacagag gattctacgc agacaaacaa
     5761 tcaacgtttg cgccgagctt cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa
     5821 gacagcatcc ttgaacaagg acaattaaca gtcaacaaat aaaaacgcaa aagaaaatgc
     5881 cgatatgcta tcggcattac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6361 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6721 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6781 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6841 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6901 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6961 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7021 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7081 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7141 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7201 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7261 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7321 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7381 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7441 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7501 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7561 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7621 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7681 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     7741 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     7801 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     7861 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     7921 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     7981 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     8041 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     8101 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     8161 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     8221 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     8281 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg ttccaactcg
     8341 cttaattgcg agtttttatt tcgtttattt caattaaggt aactaaaaaa ctccttatac
     8401 ctcgaggaaa cgccggcgaa agcggccgca aacgtcgacg aaagcgatcg caaattaatt
     8461 aaaaacgcgc gcgaaacctc agcaaagacg tcaaaaccgg taaacctagg aaaactagta
     8521 aaaagc
//
