LOCUS       40924_22518        8663 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGPro5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8663)
  AUTHORS   Thilmony R.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-AUG-2011) Crop Improvement and Utilization research 
            Unit, USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   2  (bases 1 to 8663)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8663)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (17-AUG-2011) Crop Improvement and Utilization research Unit, 
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..25)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    264..285
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        300..330
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    338..354
                     /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)."
     primer_bind     362..378
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        399..417
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    430..537
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     sig_peptide     540..620
                     /label=GRP signal sequence
                     /note="signal sequence from Oryza sativa glycine-rich cell
                     wall protein"
     intron          630..819
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             814..2637
                     /codon_start=1
                     /label=GUSPlus(TM)
                     /note="beta-glucuronidase"
                     /translation="LQNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYESKLT
                     DTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAIVYV
                     NGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGLGKV
                     IRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAETVK
                     VSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEPFGV
                     RTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFRTAH
                     YPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHHQDV
                     LRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLFVMA
                     TPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITEYGA
                     DTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRVQGN
                     KKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     CDS             2644..2661
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2693..2945
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2955..2988
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    2955..2988
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        2995..4704
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin
                     gene RUBQ2"
     CDS             4725..5747
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     protein_bind    complement(5782..5815)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5830..6004
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    complement(6082..6106)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(6116..6138)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     rep_origin      complement(6273..6708)
                     /direction=LEFT
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6999..7811
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7985..8573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaggcc ttcatgttct ttcctgcgtt atcccctgat
       61 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
      121 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
      181 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
      241 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
      301 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
      361 acaggaaaca gctatgacca tgattacgcc aagctcggaa ttaaccctca ctaaagggaa
      421 caaaagctgg agctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg
      481 caggaattcg atatcaagct tatcgatacc gtcgacctcg agggggggcc cggtacccca
      541 tggctactac taagcatttg gctcttgcca tccttgtcct ccttagcatt ggtatgacca
      601 ccagtgcaag aaccctccta gatctgaggg taaatttcta gtttttctcc ttcattttct
      661 tggttaggac ccttttctct ttttattttt ttgagctttg atctttcttt aaactgatct
      721 attttttaat tgattggtta tggtgtaaat attacatagc tttaactgat aatctgatta
      781 ctttatttcg tgtgtctatg atgatgatga tagttacaga accgacgaac tagtctgtac
      841 ccgatcaaca ccgagacccg tggcgtcttc gacctcaatg gcgtctggaa cttcaagctg
      901 gactacggga aaggactgga agagaagtgg tacgaaagca agctgaccga cactattagt
      961 atggccgtcc caagcagtta caatgacatt ggcgtgacca aggaaatccg caaccatatc
     1021 ggatatgtct ggtacgaacg tgagttcacg gtgccggcct atctgaagga tcagcgtatc
     1081 gtgctccgct tcggctctgc aactcacaaa gcaattgtct atgtcaatgg tgagctggtc
     1141 gtggagcaca agggcggatt cctgccattc gaagcggaaa tcaacaactc gctgcgtgat
     1201 ggcatgaatc gcgtcaccgt cgccgtggac aacatcctcg acgatagcac cctcccggtg
     1261 gggctgtaca gcgagcgcca cgaagagggc ctcggaaaag tcattcgtaa caagccgaac
     1321 ttcgacttct tcaactatgc aggcctgcac cgtccggtga aaatctacac gaccccgttt
     1381 acgtacgtcg aggacatctc ggttgtgacc gacttcaatg gcccaaccgg gactgtgacc
     1441 tatacggtgg actttcaagg caaagccgag accgtgaaag tgtcggtcgt ggatgaggaa
     1501 ggcaaagtgg tcgcaagcac cgagggcctg agcggtaacg tggagattcc gaatgtcatc
     1561 ctctgggaac cactgaacac gtatctctac cagatcaaag tggaactggt gaacgacgga
     1621 ctgaccatcg atgtctatga agagccgttc ggcgtgcgga ccgtggaagt caacgacggc
     1681 aagttcctca tcaacaacaa accgttctac ttcaagggct ttggcaaaca tgaggacact
     1741 cctatcaacg gccgtggctt taacgaagcg agcaatgtga tggatttcaa tatcctcaaa
     1801 tggatcggcg ccaacagctt ccggaccgca cactatccgt actctgaaga gttgatgcgt
     1861 cttgcggatc gcgagggtct ggtcgtgatc gacgagactc cggcagttgg cgtgcacctc
     1921 aacttcatgg ccaccacggg actcggcgaa ggcagcgagc gcgtcagtac ctgggagaag
     1981 attcggacgt ttgagcacca tcaagacgtt ctccgtgaac tggtgtctcg tgacaagaac
     2041 catccaagcg tcgtgatgtg gagcatcgcc aacgaggcgg cgactgagga agagggcgcg
     2101 tacgagtact tcaagccgtt ggtggagctg accaaggaac tcgacccaca gaagcgtccg
     2161 gtcacgatcg tgctgtttgt gatggctacc ccggagacgg acaaagtcgc cgaactgatt
     2221 gacgtcatcg cgctcaatcg ctataacgga tggtacttcg atggcggtga tctcgaagcg
     2281 gccaaagtcc atctccgcca ggaatttcac gcgtggaaca agcgttgccc aggaaagccg
     2341 atcatgatca ctgagtacgg cgcagacacc gttgcgggct ttcacgacat tgatccagtg
     2401 atgttcaccg aggaatatca agtcgagtac taccaggcga accacgtcgt gttcgatgag
     2461 tttgagaact tcgtgggtga gcaagcgtgg aacttcgcgg acttcgcgac ctctcagggc
     2521 gtgatgcgcg tccaaggaaa caagaagggc gtgttcactc gtgaccgcaa gccgaagctc
     2581 gccgcgcacg tctttcgcga gcgctggacc aacattccag atttcggcta caagaacgct
     2641 agccatcacc atcaccatca cgtgtgaatt ggtgaccagc tcgaatttcc ccgatcgttc
     2701 aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat
     2761 catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt
     2821 atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga
     2881 aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact
     2941 agatcggggg taccataact tcgtatagca tacattatac gaagttatgc atgcattcgg
     3001 gtcaaggcgg aagccagcgc gccaccccac gtcagcaaat acggaggcgc ggggttgacg
     3061 gcgtcacccg gtcctaacgg cgaccaacaa accagccaga agaaattaca gtaaaaaaaa
     3121 agtaaattgc actttgatcc accttttatt acctaagtct caatttggat cacccttaaa
     3181 cctatctttt caatttgggc cgggttgtgg tttggactac catgaacaac ttttcgtcat
     3241 gtctaacttc cctttcagca aacatatgaa ccatatatag aggagatcgg ccgtatacta
     3301 gagctgatgt gtttaaggtc gttgattgca cgagaaaaaa aaatccaaat cgcaacaata
     3361 gcaaatttat ctggttcaaa gtgaaaagat atgtttaaag gtagtccaaa gtaaaactta
     3421 tagataataa aatgtggtcc aaagcgtaat tcactcaaaa aaaatcaacg agacgtgtac
     3481 caaacggaga caaacggcat cttctcgaaa tttcccaacc gctcgctcgc ccgcctcgtc
     3541 ttcccggaaa ccgcggtggt ttcagcgtgg cggattctcc aagcagacgg agacgtcacg
     3601 gcacgggact cctcccacca cccaaccgcc ataaatacca gccccctcat ctcctctcct
     3661 cgcatcagct ccacccccga aaaatttctc cccaatctcg cgaggctctc gtcgtcgaat
     3721 cgaatcctct cgcgtcctca aggtacgctg cttctcctct cctcgcttcg tttcgattcg
     3781 atttcggacg ggtgaggttg ttttgttgct agatccgatt ggtggttagg gttgtcgatg
     3841 tgattatcgt gagatgttta ggggttgtag atctgatggt tgtgatttgg gcacggttgg
     3901 ttcgataggt ggaatcgtgg ttaggttttg ggattggatg ttggttctga tgattggggg
     3961 gaatttttac ggttagatga attgttggat gattcgattg gggaaatcgg tgtagatctg
     4021 ttggggaatt gtggaactag tcatgcctga gtgattggtg cgatttgtag cgtgttccat
     4081 cttgtaggcc ttgttgcgag catgttcaga tctactgttc cgctcttgat tgagttattg
     4141 gtgccttggg ttggtgcaaa cacaggcttt aatatgttat atctgttttg tgtttgatgt
     4201 agatctgtag ggtagttctt cttagacatg gttcaattat gtagcttgtg cgtttcgatt
     4261 tgatttcata tgttcacaga ttagataatg atgaactctt ttaattaatt gtcaatggta
     4321 aataggaagt cttgtcgcta tatctgtcat aatgatctca tgttactatc tgccagtaat
     4381 ttatgctaag aactatatta gaatatcatg ttacaatctg tagtaatatc atgttacaat
     4441 ctgtagttca tctatataat ctattgtggt aatttctttt tactatctgt gtgaagatta
     4501 ttgccactag ttcattctac ttatttctga agttcaggat acgtgtgctg ttactaccta
     4561 tctgaataca tgtgtgatgt gcctgttact atctttttga atacatgtat gttctgttgg
     4621 aatatgtttg ctgtttgatc cgttgttgtg tccttaatct tgtgctagtt cttaccctat
     4681 ctgtttggtg attatttctt gcagattcag atcgctagcc caccatgaaa aagcctgaac
     4741 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     4801 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     4861 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     4921 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggag tttagcgaga
     4981 gcctgaccta ttgcatctcc cgccgttcac agggtgtcac gttgcaagac ctgcctgaaa
     5041 ccgaactgcc cgctgttcta caaccggtcg cggaggctat ggatgcgatc gctgcggccg
     5101 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     5161 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     5221 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     5281 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     5341 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     5401 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     5461 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccacgactc cgggcgtata
     5521 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     5581 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     5641 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     5701 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaagaaatag agtagatgcc
     5761 gaccggatct gtcgaactag cataacttcg tatagcatac attatacgaa gttattcgac
     5821 aagctcgagt ttctccataa taatgtgtga gtagttccca gataagggaa ttagggttcc
     5881 tatagggttt cgctcatgtg ttgagcatat aagaaaccct tagtatgtat ttgtatttgt
     5941 aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagta ctaaaatcca
     6001 gatcccccga attaattcgg cgttaattca gtacattaaa aacgtccgca atgtgttatt
     6061 aagttgtcta agcgtcaatt tgtttacacc acaatatatc ctgccaccgt taacgttaca
     6121 ccacaatata tcctgccaag atctctaatt ccggggatcg gaaatccaga agcccgagag
     6181 gttgccgcct ttcgggcttt ttctttttca aaaaaaaaaa tttataaaac gatctgttgc
     6241 ggccggccgc cgggttgtgg gcaaaggcgc tggcgctcga cggtgggcaa ccgcttgcgg
     6301 ttgtccacgg gcggagccgg tgcgcgtagc gcattgtcca caagccaagg gcgaccaata
     6361 attgatatat atattcataa ttgaaaagct aattgaacat actacttgct gtaactactt
     6421 gccggagcga ggggtgtttg caagctgttg atctgaaagg gctattagcg ttctcacgtg
     6481 cctttttgat tagcgatttc acgtgacctt attagcgatt tcacgtactc cgattagcga
     6541 tttcacgtac cctgattagc gatttcacgt ggatagtttt tggagcgggc cggaaagccc
     6601 cgtgaatcaa ggctttgcgg ggcattagcg gtttcacgtg gataactacc ctctatccac
     6661 aggcttccgg ggataaaaaa gcccgctcga cggcgggctg ttggatgggg atcgcctgaa
     6721 tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg
     6781 tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa
     6841 gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccacgttg
     6901 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     6961 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     7021 cgtcttgctc aaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     7081 gggctcgcga taatgtcggg caatcaggtg cgacaatcta ccgattgtat gggaagcccg
     7141 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgtt gttacagatg
     7201 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     7261 tccgtactcc tgatgatgca tggttactca ccactgcgat cccagggaaa acagcattcc
     7321 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     7381 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     7441 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     7501 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     7561 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     7621 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     7681 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     7741 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     7801 atgagttttt ctaatcacta gaccaatgtt acacatatat actttagatt gatttaaaac
     7861 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7921 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7981 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     8041 taccagcggt ggtttgtttg ccggatcaag agctaccaac tcttcttccg aaggtaactg
     8101 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     8161 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     8221 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8281 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8341 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8401 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8461 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8521 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8581 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgagatctc
     8641 aaacaaacac atacagcgac tta
//
