LOCUS       40924_964        6312 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY B donor, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6312)
  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  JOURNAL   Unpublished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 6312)
  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 6312)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6312)
  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            "Unpublished"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..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    647..702
                     /label=A118 attB
                     /note="A118 attB"
     misc_feature    1040..1094
                     /label=TP901 attB
                     /note="TP901 attB"
     misc_feature    1116..1140
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        1299..1327
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1516..2046
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     misc_feature    2125..2230
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(2253..2269)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2277..2293)
                     /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(2301..2331)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2346..2367)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2600..3045
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3046..4464
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     rep_origin      complement(4565..5153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5327..6184)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6185..6289)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgttaacaa aagtactttt agatctgtaa ctttttcgga
      661 tcaagctatg aaggacgcaa agagggaact aaacacttaa ttacgcgttt tataacggtc
      721 ctaaggtagc gaaaaatagg gataacaggg taatcatcct cgaggaaacg ccggcgaaag
      781 cggccgcaaa cgtcgacgaa agcgatcgca aattaattaa aaacgcgcgc gaaacctcag
      841 caaagacgtc aaaaccggta aacctaggaa aactagtaaa aagcttaaag ctagcaaaca
      901 attgaaaggg cccaaaaacg ttaaatctag aaggcctccc gggaaaggat ccaaagaatt
      961 caaagtcgac aaaatcgata aagagctcaa acctgcagga aaggtaccaa accatggaaa
     1021 ggcgcgccgt ttaaacaaac tgataattgc caacacaatt aacatctcaa tcaaggtaaa
     1081 tgctttttgc ttttaaagtt taaactatca gtgtttgaca ggatatattg gcgggtaaac
     1141 ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta
     1201 tccgttcgtc catttgtatg tgcatgccaa ccacagggtt cccctcggga gaaacgtacg
     1261 gacgcacacc gtggaaacgg atgaaggcac gaacccagtt gacataagcc tgttcggttc
     1321 gtaaactgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     1381 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgtac
     1441 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     1501 tatggagcag caacgatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct
     1561 aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac
     1621 caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc
     1681 acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca
     1741 ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt
     1801 ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccgga
     1861 gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac
     1921 gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct
     1981 ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc
     2041 gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt gttcggtaaa
     2101 ttgtcacaac accgccgtac gaaaggtcaa attgggtata cccatttggg cctagtctag
     2161 ccggcatggc gcattacagc aatacgcaat ttaaatgcgc ctagcgcatt ttcccgacct
     2221 taatgcgcct aaacgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     2281 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     2341 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     2401 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     2461 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2521 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2581 aacgcaggaa agaacatggc acatatacct gccgttcact attatttagt gaaatgagat
     2641 attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc aacagaaact
     2701 ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta ggcccgtagt
     2761 ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca gttgcaatcc
     2821 aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt tactgataaa
     2881 gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca ccccttacat
     2941 attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag gagggctgga
     3001 agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa catcaaaaag
     3061 tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg aggcgcaact
     3121 caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg catttcccat
     3181 attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa atatcaagtt
     3241 cctgagttcg attcgtccac aattaaaaat atctcttctg caaaaggtct ggacgtttgg
     3301 gacagctggc cattacaaaa cgctgacggc actgtcgcaa actatcacgg ctaccacatc
     3361 gtctttgcat tagccggaga tcctaaaaat gcggatgaca cctcgattta tatgttctat
     3421 caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt ctttaaagac
     3481 agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga atggtcaggt
     3541 tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt ctccggtaaa
     3601 cattacggca aacaaacact gacaactgca caagtcaacg tatcagcatc agacagctct
     3661 ttgaacatca acggtgtaga ggactataaa tcaatctttg acggtgacgg aaaaacgtat
     3721 caaaatgtcc agcagttcat cgacgaaggc aactacagct caggcgacaa ccatacgctg
     3781 agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga agcaaacact
     3841 ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata ctatggcaaa
     3901 agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg
     3961 gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta cacactgaaa
     4021 aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga acgcgcgaac
     4081 gtctttaaaa tgaacggcaa atggtatctg ttcactgact cccgcggatc aaaaatgacg
     4141 attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa ttctttaact
     4201 ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct tgatcctaac
     4261 gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg
     4321 attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac gtttgcgccg
     4381 agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag catccttgaa
     4441 caaggacaat taacagtcaa caaataaaaa cgcaaaagaa aatgccgata tgctatcggc
     4501 attacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4561 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4681 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4741 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4801 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4861 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4921 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4981 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5041 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5101 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5161 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5221 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5281 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5341 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5401 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5461 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5521 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5581 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5641 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5701 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5761 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5821 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5881 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5941 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6001 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6061 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6121 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6181 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6241 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6301 gaaaagtgcc ac
//
