LOCUS       40924_939        8899 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY B donor eGFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8899)
  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  JOURNAL   Unpublished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 8899)
  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 8899)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8899)
  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            "Unpublished"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..8899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      complement(8..219)
                     /label=CaMV 35S terminator
                     /note="CaMV 35S terminator"
                     /regulatory_class="terminator"
     CDS             complement(238..954)
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             complement(959..1188)
                     /codon_start=1
                     /gene="ubq"
                     /product="ubiquitin monomer"
                     /label=ubq
                     /note="ubiquitin monomer confers enhanced expression in 
                     translational fusions"
                     /protein_id="AWK49545.1"
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLADYNIQKESTLHLVLRLRGG"
     gene            complement(959..1188)
                     /gene="ubq"
                     /label=ubq
     regulatory      complement(1189..2907)
                     /label=potato ubiquitin 7 promoter and 5' intron
                     /note="potato ubiquitin 7 promoter and 5' intron"
                     /regulatory_class="promoter"
     protein_bind    2796..2807
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     misc_feature    2925..2979
                     /label=TP901 attB
                     /note="TP901 attB"
     misc_feature    3001..3025
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        3184..3212
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3401..3931
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     misc_feature    4010..4115
                     /label=ParA MRS
                     /note="ParA MRS"
     primer_bind     complement(4138..4154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4162..4178)
                     /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(4186..4216)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4231..4252)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4485..4930
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4931..6349
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     rep_origin      complement(6450..7038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7212..8069)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8070..8174)
                     /label=AmpR promoter
     rep_origin      complement(8200..8655)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     8797..8813
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8844..8899
                     /label=A118 attB
                     /note="A118 attB"
ORIGIN
        1 acgcgccggt cactggattt tggttttagg aattagaaat tttattgata gaagtatttt
       61 acaaatacaa atacatacta agggtttctt atatgctcaa cacatgagcg aaaccctata
      121 agaaccctaa ttcccttatc tgggaactac tcacacatta ttctggagaa aaatagagag
      181 agatagattt gtagagagag actggtgatt tttgcggacg gtacccccaa gcttttactt
      241 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
      301 gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt
      361 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
      421 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
      481 ctgcttgtcg accatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
      541 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
      601 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
      661 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
      721 gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg
      781 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
      841 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
      901 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggcgcc
      961 acctccacgt agacggagca ccaaatggag ggtagactcc ttctggatgt tgtaatcagc
     1021 tagagtacgt ccgtcctcca actgctttcc ggcgaagata agcctttgct gatccggggg
     1081 aatcccttcc ttatcctgga tcttagcctt aacgttgtcg attgtatcag aactttccac
     1141 ctctagggtg atagtctttc cggtgagagt tttcacaaat atctgcatct gcaaattcat
     1201 aaaaacaaca atcagaacaa cgaaaaaaac ttcaatttag ggctttcaca ctccctgatt
     1261 agctgattcg agaattaaaa cctaaacaat ctaaaacacg gggaaaaaac ttcaaattta
     1321 gggctttttc atcaaaacag ccggagaaac tcaaatttag ggctttttca tcaaaacagc
     1381 cggagaaact caaattttag gccttcagaa tcaaaccgac ggaaaaacac aaattctagg
     1441 gctttttcat caaatcaacc ggaaaaactc aaatttaagg cctttttcat caaacaaatg
     1501 taaaaactct aattttaaac ctttagaacg aacgacagaa aaaaaatctc aaactagggg
     1561 ctttttcatc aaaacgaacg gggaaaacgg ccaattgtag gccttttcca tcaaaccaga
     1621 cgtaaaaaaa ctcaaatgtt aggccttttc catcaagaca accgaggaga gactcaaaag
     1681 gaagggctat aacagcgcct gattagctaa ttcgagaatc taaacataca aaaaccaaaa
     1741 atagagaaga ttctaacctt aaagagaaat ttgaaggcaa ttgagagaaa ttataataga
     1801 aatcagatag aagatgaaaa aaatgaggtg cagaatgctt cattttatag atggcatggc
     1861 cgactcaaga cgaaaccgcc tcatagggtt aaccaattat atgaggacac gggtaataat
     1921 gaggtgtagt gtaccaaaac aacacgcaaa agctaccttt cctttttaat ttttatgtgg
     1981 tggggaaaat taggagaatg aggtcgacac ttacacgctc ctttagaaaa acccagaaaa
     2041 atgtaggtca tgtaaaaaaa tatatactaa aaataactta attttatttt aggtattttt
     2101 ttattttaat atatgctcta atatagcgca tttcaagttt atcatgaaca tgaccttaaa
     2161 tataagagcc aaatattagc aaattaggat ataatactac cttgataatc gcacttaaat
     2221 cgtcccttat attatgtatt gctctttttt catattattt tgatattgtt attgacctca
     2281 tattgatatt tgttatttaa aatttgattg aaaatattta atgtaaattg agggtctatt
     2341 agaaacactt tttcttattt ctaagaggta agtattagat tccatgaaat tatattagtg
     2401 ttttattagg ttaaagttaa taatttattt tcttatgata aaaaaaaaaa aagtcagtcg
     2461 acttcactct ttacctcaaa tttattttat gtcttcttaa tgactttttt ttattggaaa
     2521 aatttagtaa tttggttata aaaatagttt atttcatgat atttaagtaa aatttgaatt
     2581 gttttacatt atcaaatata atttagtgac gtggtataat taaaaaaaaa ttgacctctt
     2641 gctttataag aaatattatt attattacta tattgcatat aaaactcaat caatgtgaat
     2701 aataaatggg tttttgtgtc tgtcattatc attatgaaaa acattgaagt tgcttactat
     2761 tcaatttggg tatatgtaat tgatgaccaa atcttgtcat cctcatcaac aaaactaatg
     2821 taaattttaa taaaactttt tgaagtgaaa ttgtcttgga tataaagtga atttaaaaaa
     2881 tcataaaatt gagatttcaa atattggcgc gccgtttaaa caaactgata attgccaaca
     2941 caattaacat ctcaatcaag gtaaatgctt tttgctttta aagtttaaac tatcagtgtt
     3001 tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat
     3061 atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca
     3121 gggttcccct cgggagaaac gtacggacgc acaccgtgga aacggatgaa ggcacgaacc
     3181 cagttgacat aagcctgttc ggttcgtaaa ctgtaatgca agtagcgtat gcgctcacgc
     3241 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
     3301 gcttgttatg actgtttttt tgtacagtct atgcctcggg catccaagca gcaagcgcgt
     3361 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagcaacga
     3421 tgttacgcag cagggcagtc gccctaaaac aaagttaggt ggctcaagta tgggcatcat
     3481 tcgcacatgt aggctcggcc ctgaccaagt caaatccatg cgggctgctc ttgatctttt
     3541 cggtcgtgag ttcggagacg tagccaccta ctcccaacat cagccggact ccgattacct
     3601 cgggaacttg ctccgtagta agacattcat cgcgcttgct gccttcgacc aagaagcggt
     3661 tgttggcgct ctcgcggctt acgttctgcc caagtttgag cagccgcgta gtgagatcta
     3721 tatctatgat ctcgcagtct ccggagagca ccggaggcag ggcattgcca ccgcgctcat
     3781 caatctcctc aagcatgagg ccaacgcgct tggtgcttat gtgatctacg tgcaagcaga
     3841 ttacggtgac gatcccgcag tggctctcta tacaaagttg ggcatacggg aagaagtgat
     3901 gcactttgat atcgacccaa gtaccgccac ctaacaattc gttcaagccg agatcggctt
     3961 cccggccgcg gagttgttcg gtaaattgtc acaacaccgc cgtacgaaag gtcaaattgg
     4021 gtatacccat ttgggcctag tctagccggc atggcgcatt acagcaatac gcaatttaaa
     4081 tgcgcctagc gcattttccc gaccttaatg cgcctaaacg cgcttggcgt aatcatggtc
     4141 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4201 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4261 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4321 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4441 acggttatcc acagaatcag gggataacgc aggaaagaac atggcacata tacctgccgt
     4501 tcactattat ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca
     4561 actttatgcc catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga
     4621 ttttttagtt ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa
     4681 gaggaaaata gaccagttgc aatccaaacg agagtctaat agaatgaggt cgaaaagtaa
     4741 atcgcgcggg tttgttactg ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg
     4801 tatactttgg cgtcacccct tacatatttt aggtcttttt ttattgtgcg taactaactt
     4861 gccatcttca aacaggaggg ctggaagaag cagaccgcta acacagtaca taaaaaagga
     4921 gacatgaacg atgaacatca aaaagtttgc aaaacaagca acagtattaa cctttactac
     4981 cgcactgctg gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc aaaagccata
     5041 taaggaaaca tacggcattt cccatattac acgccatgat atgctgcaaa tccctgaaca
     5101 gcaaaaaaat gaaaaatatc aagttcctga gttcgattcg tccacaatta aaaatatctc
     5161 ttctgcaaaa ggtctggacg tttgggacag ctggccatta caaaacgctg acggcactgt
     5221 cgcaaactat cacggctacc acatcgtctt tgcattagcc ggagatccta aaaatgcgga
     5281 tgacacctcg atttatatgt tctatcaaaa agtcggcgaa acttctattg acagctggaa
     5341 aaacgctggc cgcgtcttta aagacagcga caaattcgat gcaaatgatt ctatcctaaa
     5401 agaccaaaca caagaatggt caggttcagc cacatttaca tctgacggaa aaatccgttt
     5461 attctacact gatttctccg gtaaacatta cggcaaacaa acactgacaa ctgcacaagt
     5521 caacgtatca gcatcagaca gctctttgaa catcaacggt gtagaggact ataaatcaat
     5581 ctttgacggt gacggaaaaa cgtatcaaaa tgtccagcag ttcatcgacg aaggcaacta
     5641 cagctcaggc gacaaccata cgctgagaga tcctcactac gtagaagata aaggccacaa
     5701 atacttagta tttgaagcaa acactggaac tgaagatggc taccaaggcg aagaatcttt
     5761 atttaacaaa gcatactatg gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact
     5821 tctgcaaagc gataaaaaac gcacggctga gttagcaaac ggcgctctcg gtatgattga
     5881 gctaaacgat gattacacac tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt
     5941 aacagatgaa attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt atctgttcac
     6001 tgactcccgc ggatcaaaaa tgacgattga cggcattacg tctaacgata tttacatgct
     6061 tggttatgtt tctaattctt taactggccc atacaagccg ctgaacaaaa ctggccttgt
     6121 gttaaaaatg gatcttgatc ctaacgatgt aacctttact tactcacact tcgctgtacc
     6181 tcaagcgaaa ggaaacaatg tcgtgattac aagctatatg acaaacagag gattctacgc
     6241 agacaaacaa tcaacgtttg cgccgagctt cctgctgaac atcaaaggca agaaaacatc
     6301 tgttgtcaaa gacagcatcc ttgaacaagg acaattaaca gtcaacaaat aaaaacgcaa
     6361 aagaaaatgc cgatatgcta tcggcattac atgtgagcaa aaggccagca aaaggccagg
     6421 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6481 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6541 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6601 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6661 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6721 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6781 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6841 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6901 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6961 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7021 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7081 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7141 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7201 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7261 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7321 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7381 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7441 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7501 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7561 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7621 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7681 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7741 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7801 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7861 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7921 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7981 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8041 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8101 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8161 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     8221 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     8281 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     8341 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     8401 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     8461 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     8521 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     8581 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     8641 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     8701 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     8761 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     8821 ttaacaaaag tacttttaga tctgtaactt tttcggatca agctatgaag gacgcaaaga
     8881 gggaactaaa cacttaatt
//
