LOCUS       40924_974        7945 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P donor bar, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7945)
  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  JOURNAL   Unpublished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 7945)
  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 7945)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7945)
  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            "Unpublished"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..7945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..253)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(288..836)
                     /codon_start=1
                     /label=BlpR
                     /note="phosphinothricin acetyltransferase"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(887..1066)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     complement(1473..1489)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1589..1644
                     /label=A118 attP
                     /note="A118 attP"
     misc_feature    1681..1705
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2037..2828
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    3023..3128
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(3151..3167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3175..3191)
                     /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(3199..3229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3244..3265)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3498..3943
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3944..5362
                     /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(5463..6051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6225..7082)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7083..7187)
                     /label=AmpR promoter
     rep_origin      complement(7213..7668)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7810..7826
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7835..7899
                     /label=TP901 attP
                     /note="TP901 attP"
ORIGIN
        1 gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt
       61 tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca
      121 taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt
      181 atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa
      241 tgtttgaacg atcggggaaa ttcgagctga tcctctaggg gtcatcagat ctcggtgacg
      301 ggcaggaccg gacggggcgg taccggcagg ctgaagtcca gctgccagaa acccacgtca
      361 tgccagttcc cgtgcttgaa gccggccgcc cgcagcatgc cgcggggggc atatccgagc
      421 gcctcgtgca tgcgcacgct cgggtcgttg ggcagcccga tgacagcgac cacgctcttg
      481 aagccctgtg cctccaggga cttcagcagg tgggtgtaga gcgtggagcc cagtcccgtc
      541 cgctggtggc ggggggagac gtacacggtc gactcggccg tccagtcgta ggcgttgcgt
      601 gccttccagg ggcccgcgta ggcgatgccg gcgacctcgc cgtccacctc ggcgacgagc
      661 cagggatagc gctcccgcag acggacgagg tcgtccgtcc actcctgcgg ttcctgcggc
      721 tcggtacgga agttgaccgt gcttgtctcg atgtagtggt tgacgatggt gcagaccgcc
      781 ggcatgtccg cctcggtggc acggcggatg tcggccgggc gtcgttctgg gctcatggta
      841 gatcccccta gagtcgagat ctggattgag agtgaatatg agactctaat tggataccga
      901 ggggaattta tggaacgtca gtggagcatt tttgacaaga aatatttgct agctgatagt
      961 gaccttaggc gacttttgaa cgcgcaataa tggtttctga cgtatgtgct tagctcatta
     1021 aactccagaa acccgcggct gagtggctcc ttcaacgttg cggttctgtc agttccaaac
     1081 gtaaaacggc ttgtcccgcg tcatcggcgg gggtcataac gtgactccct taattctccg
     1141 ctcatgatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     1201 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg
     1261 ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct
     1321 ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     1381 gtcagcaccg tttctgcgga ctggctttct acgtgttccg cttcctttag cagcccttgc
     1441 gccctgagtg cttgcggcag cgtgaagctt atcgataccg tcgacctcga gggggggccc
     1501 ggtaccgcgg ccgcttaatt aaggcgcgcc tttataacgg tcctaaggta gcgaaaaata
     1561 gggataacag ggtaatttta cgcgtaaatg tttagttcct cgttttctct cgttggaaga
     1621 agaagaaacg agaaactaaa attaaaaagt acttttagat ctgtttaaac tatcagtgtt
     1681 tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat
     1741 atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca
     1801 gggttcccct cgggagaaac gtacgctcgc gttggcagca tcacccataa ttgtggtttc
     1861 aaaatcggct ccgtcgatac tatgttatac gccaactttg aaaacaactt tgaaaaagct
     1921 gttttctggt atttaaagtt ttagaatgca aggaacagtg aattggagtt cgtcttgtta
     1981 taattagctt cttggggtat ctttaaatac tgtagaaaag aggaaggaaa taataaatgg
     2041 ctaaaatgag aatatcaccg gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag
     2101 atacggaagg aatgtctcct gctaaggtat ataagctggt gggagaaaat gaaaacctat
     2161 atttaaaaat gacggacagc cggtataaag ggaccaccta tgatgtggaa cgggaaaagg
     2221 acatgatgct atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac cttgaacggc
     2281 atgatggctg gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt
     2341 atgaagatga acaaagccct gaaaagatta tcgagctgta tgcggagtgc atcaggctct
     2401 ttcactccat cgacatatcg gattgtccct atacgaatag cttagacagc cgcttagccg
     2461 aattggatta cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag
     2521 acactccatt taaagatccg cgcgagctgt atgatttttt aaagacggaa aagcccgaag
     2581 aggaacttgt cttttcccac ggcgacctgg gagacagcaa catctttgtg aaagatggca
     2641 aagtaagtgg ctttattgat cttgggagaa gcggcagggc ggacaagtgg tatgacattg
     2701 ccttctgcgt ccggtcgatc agggaggata ttggggaaga acagtatgtc gagctatttt
     2761 ttgacttact ggggatcaag cctgattggg agaaaataaa atattatatt ttactggatg
     2821 aattgtttta gtacctagat gtggcgcaac gatgccggcg acaagcagga gcgcaccgac
     2881 ttcttccgca tcaagtgttt tggctctcag gccgaggccc acggcaagta tttgggcaag
     2941 gggtcgctgg tattcgtgca gggcaagatt cggaatacca agtacgagaa ggacggccag
     3001 acggtctacg ggacgtacga aaggtcaaat tgggtatacc catttgggcc tagtctagcc
     3061 ggcatggcgc attacagcaa tacgcaattt aaatgcgcct agcgcatttt cccgacctta
     3121 atgcgcctaa acgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3181 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3241 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3301 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3361 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3421 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3481 cgcaggaaag aacatggcac atatacctgc cgttcactat tatttagtga aatgagatat
     3541 tatgatattt tctgaattgt gattaaaaag gcaactttat gcccatgcaa cagaaactat
     3601 aaaaaataca gagaatgaaa agaaacagat agatttttta gttctttagg cccgtagtct
     3661 gcaaatcctt ttatgatttt ctatcaaaca aaagaggaaa atagaccagt tgcaatccaa
     3721 acgagagtct aatagaatga ggtcgaaaag taaatcgcgc gggtttgtta ctgataaagc
     3781 aggcaagacc taaaatgtgt aaagggcaaa gtgtatactt tggcgtcacc ccttacatat
     3841 tttaggtctt tttttattgt gcgtaactaa cttgccatct tcaaacagga gggctggaag
     3901 aagcagaccg ctaacacagt acataaaaaa ggagacatga acgatgaaca tcaaaaagtt
     3961 tgcaaaacaa gcaacagtat taacctttac taccgcactg ctggcaggag gcgcaactca
     4021 agcgtttgcg aaagaaacga accaaaagcc atataaggaa acatacggca tttcccatat
     4081 tacacgccat gatatgctgc aaatccctga acagcaaaaa aatgaaaaat atcaagttcc
     4141 tgagttcgat tcgtccacaa ttaaaaatat ctcttctgca aaaggtctgg acgtttggga
     4201 cagctggcca ttacaaaacg ctgacggcac tgtcgcaaac tatcacggct accacatcgt
     4261 ctttgcatta gccggagatc ctaaaaatgc ggatgacacc tcgatttata tgttctatca
     4321 aaaagtcggc gaaacttcta ttgacagctg gaaaaacgct ggccgcgtct ttaaagacag
     4381 cgacaaattc gatgcaaatg attctatcct aaaagaccaa acacaagaat ggtcaggttc
     4441 agccacattt acatctgacg gaaaaatccg tttattctac actgatttct ccggtaaaca
     4501 ttacggcaaa caaacactga caactgcaca agtcaacgta tcagcatcag acagctcttt
     4561 gaacatcaac ggtgtagagg actataaatc aatctttgac ggtgacggaa aaacgtatca
     4621 aaatgtccag cagttcatcg acgaaggcaa ctacagctca ggcgacaacc atacgctgag
     4681 agatcctcac tacgtagaag ataaaggcca caaatactta gtatttgaag caaacactgg
     4741 aactgaagat ggctaccaag gcgaagaatc tttatttaac aaagcatact atggcaaaag
     4801 cacatcattc ttccgtcaag aaagtcaaaa acttctgcaa agcgataaaa aacgcacggc
     4861 tgagttagca aacggcgctc tcggtatgat tgagctaaac gatgattaca cactgaaaaa
     4921 agtgatgaaa ccgctgattg catctaacac agtaacagat gaaattgaac gcgcgaacgt
     4981 ctttaaaatg aacggcaaat ggtatctgtt cactgactcc cgcggatcaa aaatgacgat
     5041 tgacggcatt acgtctaacg atatttacat gcttggttat gtttctaatt ctttaactgg
     5101 cccatacaag ccgctgaaca aaactggcct tgtgttaaaa atggatcttg atcctaacga
     5161 tgtaaccttt acttactcac acttcgctgt acctcaagcg aaaggaaaca atgtcgtgat
     5221 tacaagctat atgacaaaca gaggattcta cgcagacaaa caatcaacgt ttgcgccgag
     5281 cttcctgctg aacatcaaag gcaagaaaac atctgttgtc aaagacagca tccttgaaca
     5341 aggacaatta acagtcaaca aataaaaacg caaaagaaaa tgccgatatg ctatcggcat
     5401 tacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5461 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5521 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5581 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5641 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5701 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5761 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5821 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5881 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     5941 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6001 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6061 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6121 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6181 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6241 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6301 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6361 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6421 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6481 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     6541 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     6601 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     6661 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6721 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6781 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6841 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6901 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6961 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     7021 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     7081 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     7141 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     7201 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     7261 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     7321 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     7381 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     7441 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     7501 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     7561 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     7621 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     7681 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     7741 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     7801 cacgacgttg taaaacgacg gccagtgagc gcgttccaac tcgcttaatt gcgagttttt
     7861 atttcgttta tttcaattaa ggtaactaaa aaactcctta tacctcgaca aaatcgataa
     7921 agagctcaaa cctgcaggaa ttccc
//
