LOCUS       40924_979       10247 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P donor luc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10247)
  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  JOURNAL   Unpublished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 10247)
  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 10247)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10247)
  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            "Unpublished"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..10247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    51..72
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        87..117
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    125..141
                     /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)."
     primer_bind     149..165
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(204..456)
                     /label=nos terminator
                     /note="nos terminator"
                     /regulatory_class="terminator"
     terminator      204..456
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(514..2163)
                     /label=luciferase
                     /note="firefly luciferase"
     CDS             complement(2170..2400)
                     /codon_start=1
                     /gene="ubq"
                     /product="ubiquitin monomer"
                     /label=ubq
                     /note="potato ubiquitin monomer; translational fusion
                     confers enhanced expression"
                     /protein_id="AWK49540.1"
                     /translation="MQIFVKTLTGKTITLEVEGSDTIDNVKAEIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLADYNIQKESTLHLVLRLRGGG"
     gene            complement(2170..2400)
                     /gene="ubq"
                     /label=ubq
     regulatory      complement(2401..3779)
                     /label=potato 409S ubiquitin promoter and 5' intron
                     /note="potato 409S ubiquitin promoter and 5' intron"
                     /regulatory_class="promoter"
     misc_feature    3887..3942
                     /label=A118 attP
                     /note="A118 attP"
     misc_feature    3979..4003
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4335..5126
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    5321..5426
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(5449..5465)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5473..5489
                     /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(5497..5527)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5542..5563)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5796..6241
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             6242..7660
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     rep_origin      complement(7761..8349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8523..9380)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9381..9485)
                     /label=AmpR promoter
     rep_origin      complement(9511..9966)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     10108..10124
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    10133..10197
                     /label=TP901 attP
                     /note="TP901 attP"
ORIGIN
        1 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
       61 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
      121 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgaattc
      181 tcatgtttga cagcttatca tcggatctag taacatagat gacaccgcgc gcgataattt
      241 atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac
      301 tctaatcata aaaacccatc tcataaataa cgtcatgcat tacatgttaa ttattacatg
      361 cttaacgtaa ttcaacagaa attatatgat aatcatcgca agaccggcaa caggattcaa
      421 tcttaagaaa ctttattgcc aaatgtttga acgatctgca ggtcgacgga tcagatctcg
      481 gtgacgggca ggaccggacg gggcggtacc ttacacggcg atctttccgc ccttcttggc
      541 ctttatgagg atctctctga tttttcttgc gtcgagtttt ccggtaagac ctttcggtac
      601 ttcgtccaca aacacaactc ctccgcgcaa ctttttcgcg gttgttactt gactggcgac
      661 gtaatccacg atctcttttt ccgtcatcgt ctttccgtgc tccaaaacaa caacggcggc
      721 gggaagttca ccggcgtcat cgtcgggaag acctgcgaca cctgcgtcga agatgttggg
      781 gtgttggagc aagatggatt ccaattcagc gggagccacc tgatagcctt tgtacttaat
      841 cagagacttc aggcggtcaa cgatgaagaa gtgttcgtct tcgtcccagt aagctatgtc
      901 tccagaatgt agccatccat ccttgtcaat caaggcgttg gtcgcttccg gattgtttac
      961 ataaccggac ataatcatag gacctctcac acacagttcg cctctttgat taacgcccag
     1021 cgttttcccg gtatccagat ccacaacctt cgcttcaaaa aatggaacaa ctttaccgac
     1081 cgcgcccggt ttatcatccc cctcgggtgt aatcagaata gctgatgtag tctcagtgag
     1141 cccatatcct tgcctgatac ctggcagatg gaacctcttg gcaaccgctt ccccgacttc
     1201 cttagagagg ggagcgccac cagaagcaat ttcgtgtaaa ttagataaat cgtatttgtc
     1261 aatcagagtg cttttggcga agaaggagaa tagggttggc accagcagcg cactttgaat
     1321 cttgtaatcc tgaaggctcc tcagaaacag ctcttcttca aatctataca ttaagacgac
     1381 tcgaaatcca catatcaaat atccgagtgt agtaaacatt ccaaaaccgt gatggaatgg
     1441 aacaacactt aaaatcgcag tatccggaat gatttgattg ccaaaaatag gatctctggc
     1501 atgcgagaat ctcacgcagg cagttctatg aggcagagcg acacctttag gcagaccagt
     1561 agatccagag gagttcatga tcagtgcaat tgtcttgtcc ctatcgaagg actctggcac
     1621 aaaatcgtat tcattaaaac cgggaggtag atgagatgtg acgaacgtgt acatcgactg
     1681 aaatccctgg taatccgttt tagaatccat gataataatt ttttggatga ttgggagctt
     1741 tttttgcacg ttcaaaattt tttgcaaccc ctttttggaa acgaacacca cggtaggctg
     1801 cgaaatgccc atactgttga gcaattcacg ttcattataa atgtcgttcg cgggcgcaac
     1861 tgcaactccg ataaataacg cgcccaacac cggcataaag aattgaagag agttttcact
     1921 gcatacgacg attctgtgat ttgtattcag cccatatcgt ttcatagctt ctgccaaccg
     1981 aacggacatt tcgaagtact cagcgtaagt gatgtccacc tcgatatgtg catctgtaaa
     2041 agcaattgtt ccaggaacca gggcgtatct cttcatagcc ttatgcagtt gctctccagc
     2101 ggttccatct tccagcggat agaatggcgc cgggcctttc tttatgtttt tggcgtcttc
     2161 catggatcct ccaccaccac gtagacggag aaccaaatgg agggtagact ccttctggat
     2221 gttgtaatca gctagagtac gtccgtcctc caactgcttt ccggcgaaga taagcctttg
     2281 ctgatccggg ggaattcctt ccttatcctg aatctcagcc ttaacgttgt cgattgtatc
     2341 agaaccttcc acctctaggg tgatggtctt tccggtgaga gttttcacaa agatctgcat
     2401 ctgcaaagtc ataaaaacaa caatcggaac aacgaaaaaa actcaaatta tagggctttt
     2461 cacactccct gattaactga ttagaattaa aaccaaacaa tctaaaacac ggggaaaaaa
     2521 ctcaaatttt agggcttttt catcaaaaca accggaaaac ctcaaatttt agggctttag
     2581 aatcaaaccg acgaaaaatc acaaattcta gggctttttc atcaaatcaa ccggaaaaaa
     2641 aaaaatcaaa ttcaagacct ttttcatcaa accaccataa aaaaactcaa attctagggc
     2701 attatcatca aatcacccgg aaaaccccac attctgggcc tttagaatca aaccgacgaa
     2761 aaaactctaa ttttaaacct ttggcatcaa atcgacggaa aaaaactcaa attaggggct
     2821 ttttcatcaa aacaacggaa aaaaaactca aattttaggg ctttaacagc acctgcttag
     2881 gcggccgcct aattcgagaa tctaaacaga caaaaaccaa aaaatagaga agatttctaa
     2941 ccttgaaaga gaaatttaaa ggcaattgag agaaattata atagaagtca gatagaagat
     3001 gaaaaaatga ggtgcagaaa ggttcctttt atagatggca tggccgactc tagacgaaac
     3061 cgcctcatag ggttagccaa ttatatgatg acacgtgtaa taatgaggtg tagtgtagca
     3121 aaacaacacg caaaagctac cgtttctttt tatttatgtg gtggggaaaa ttaggagaat
     3181 gaggttgacg cttacacgct cctttagaaa aaactagaaa aatgtaggtc aatgtaaaaa
     3241 aatatactaa aaataactta attttatttt aggtattggc caaaatctgg ggaaaaaatt
     3301 tggtctgaat ttaattagaa tgatatttta tttatgttat tttatttttt aaaatatttt
     3361 tattctttta actttaatac attttaacta aaaaataaag aatatatcaa tttattttaa
     3421 aataaaaaaa tattataatt ttcttaattt tctatctaat ttttttgatc atttagcatt
     3481 ttctttttta ttaattaata tatgctctaa tatagcgctt tttaagtata ttgtgaacat
     3541 aaccttaaat ataagagttt aaatattagc aaattagggt ataatactcg ttgataatcg
     3601 cactttaatc ttttttctta tattatgtat tgctcctttt tttttacatt atttttttat
     3661 tgttattgac ctcatattga taagtatttg ttatctaaat ttgattgaaa atattttatt
     3721 taaattgagt atatattaga aacacttttt cttttttata taagaggtaa gtagcttggc
     3781 aagcttgcat gcctgcagga aaggtaccaa accatggaaa ggcgcgcctt tataacggtc
     3841 ctaaggtagc gaaaaatagg aataacaggg taattttacg cgtaaatgtt tagttcctcg
     3901 ttttctctcg ttggaagaag aagaaacgag aaactaaaat taaaaagtac ttttagatct
     3961 gtttaaacta tcagtgtttg acaggatata ttggcgggta aacctaagag aaaagagcgt
     4021 ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt
     4081 atgtgcatgc caaccacagg gttcccctcg ggagaaacgt acgctcgcgt tggcagcatc
     4141 acccataatt gtggtttcaa aatcggctcc gtcgatacta tgttatacgc caactttgaa
     4201 aacaactttg aaaaagctgt tttctggtat ttaaagtttt agaatgcaag gaacagtgaa
     4261 ttggagttcg tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag
     4321 gaaggaaata ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa
     4381 aataccgctg cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg
     4441 gagaaaatga aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg
     4501 atgtggaacg ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg
     4561 tcctgcacct tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg
     4621 tcctttgctc ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg
     4681 cggagtgcat caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct
     4741 tagacagccg cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt
     4801 gcgaaaactg ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa
     4861 agacggaaaa gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca
     4921 tctttgtgaa agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg
     4981 acaagtggta tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatt ggggaagaac
     5041 agtatgtcga gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat
     5101 attatatttt actggatgaa ttgttttagt acctagatgt ggcgcaacga tgccggcgac
     5161 aagcaggagc gcaccgactt cttccgcatc aagtgttttg gctctcaggc cgaggcccac
     5221 ggcaagtatt tgggcaaggg gtcgctggta ttcgtgcagg gcaagattcg gaataccaag
     5281 tacgagaagg acggccagac ggtctacggg acgtacgaaa ggtcaaattg ggtataccca
     5341 tttgggccta gtctagccgg catggcgcat tacagcaata cgcaatttaa atgcgcctag
     5401 cgcattttcc cgaccttaat gcgcctaaac gcgcttggcg taatcatggt catagctgtt
     5461 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5521 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5581 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5641 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5701 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5761 cacagaatca ggggataacg caggaaagaa catggcacat atacctgccg ttcactatta
     5821 tttagtgaaa tgagatatta tgatattttc tgaattgtga ttaaaaaggc aactttatgc
     5881 ccatgcaaca gaaactataa aaaatacaga gaatgaaaag aaacagatag attttttagt
     5941 tctttaggcc cgtagtctgc aaatcctttt atgattttct atcaaacaaa agaggaaaat
     6001 agaccagttg caatccaaac gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg
     6061 gtttgttact gataaagcag gcaagaccta aaatgtgtaa agggcaaagt gtatactttg
     6121 gcgtcacccc ttacatattt taggtctttt tttattgtgc gtaactaact tgccatcttc
     6181 aaacaggagg gctggaagaa gcagaccgct aacacagtac ataaaaaagg agacatgaac
     6241 gatgaacatc aaaaagtttg caaaacaagc aacagtatta acctttacta ccgcactgct
     6301 ggcaggaggc gcaactcaag cgtttgcgaa agaaacgaac caaaagccat ataaggaaac
     6361 atacggcatt tcccatatta cacgccatga tatgctgcaa atccctgaac agcaaaaaaa
     6421 tgaaaaatat caagttcctg agttcgattc gtccacaatt aaaaatatct cttctgcaaa
     6481 aggtctggac gtttgggaca gctggccatt acaaaacgct gacggcactg tcgcaaacta
     6541 tcacggctac cacatcgtct ttgcattagc cggagatcct aaaaatgcgg atgacacctc
     6601 gatttatatg ttctatcaaa aagtcggcga aacttctatt gacagctgga aaaacgctgg
     6661 ccgcgtcttt aaagacagcg acaaattcga tgcaaatgat tctatcctaa aagaccaaac
     6721 acaagaatgg tcaggttcag ccacatttac atctgacgga aaaatccgtt tattctacac
     6781 tgatttctcc ggtaaacatt acggcaaaca aacactgaca actgcacaag tcaacgtatc
     6841 agcatcagac agctctttga acatcaacgg tgtagaggac tataaatcaa tctttgacgg
     6901 tgacggaaaa acgtatcaaa atgtccagca gttcatcgac gaaggcaact acagctcagg
     6961 cgacaaccat acgctgagag atcctcacta cgtagaagat aaaggccaca aatacttagt
     7021 atttgaagca aacactggaa ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa
     7081 agcatactat ggcaaaagca catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag
     7141 cgataaaaaa cgcacggctg agttagcaaa cggcgctctc ggtatgattg agctaaacga
     7201 tgattacaca ctgaaaaaag tgatgaaacc gctgattgca tctaacacag taacagatga
     7261 aattgaacgc gcgaacgtct ttaaaatgaa cggcaaatgg tatctgttca ctgactcccg
     7321 cggatcaaaa atgacgattg acggcattac gtctaacgat atttacatgc ttggttatgt
     7381 ttctaattct ttaactggcc catacaagcc gctgaacaaa actggccttg tgttaaaaat
     7441 ggatcttgat cctaacgatg taacctttac ttactcacac ttcgctgtac ctcaagcgaa
     7501 aggaaacaat gtcgtgatta caagctatat gacaaacaga ggattctacg cagacaaaca
     7561 atcaacgttt gcgccgagct tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa
     7621 agacagcatc cttgaacaag gacaattaac agtcaacaaa taaaaacgca aaagaaaatg
     7681 ccgatatgct atcggcatta catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7741 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7801 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7861 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7921 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7981 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8041 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8101 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8161 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     8221 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8281 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8341 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8401 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     8461 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8521 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8581 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8641 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8701 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8761 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8821 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8881 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8941 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9001 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9061 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9121 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9181 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9241 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9301 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9361 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9421 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9481 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     9541 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     9601 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     9661 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     9721 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     9781 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     9841 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     9901 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     9961 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
    10021 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
    10081 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gttccaactc
    10141 gcttaattgc gagtttttat ttcgtttatt tcaattaagg taactaaaaa actccttata
    10201 cctcgaggaa acgccggcga aagcggccgc aaacgtcgac gaaagcg
//
