LOCUS       40924_22523        8588 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGPro8, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8588)
  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 8588)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8588)
  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..8588
                     /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"
     intron          555..744
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             739..2562
                     /codon_start=1
                     /label=GUSPlus(TM)
                     /note="beta-glucuronidase"
                     /translation="LQNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYESKLT
                     DTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAIVYV
                     NGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGLGKV
                     IRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAETVK
                     VSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEPFGV
                     RTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFRTAH
                     YPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHHQDV
                     LRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLFVMA
                     TPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITEYGA
                     DTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRVQGN
                     KKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     CDS             2569..2586
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2618..2870
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2880..2913
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    2880..2913
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        2920..4629
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin
                     gene RUBQ2"
     CDS             4650..5672
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     protein_bind    complement(5707..5740)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5755..5929
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    complement(6007..6031)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(6041..6063)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     rep_origin      complement(6198..6633)
                     /direction=LEFT
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6924..7736
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7910..8498
                     /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 aagcgcgcaa ttaaccctca ctaaagggaa
      421 caaaagctgg agctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg
      481 caggaattcg atatcaagct tatcgatacc gtcgacctcg agggggggcc cggtacccca
      541 tggtagatct gagggtaaat ttctagtttt tctccttcat tttcttggtt aggacccttt
      601 tctcttttta tttttttgag ctttgatctt tctttaaact gatctatttt ttaattgatt
      661 ggttatggtg taaatattac atagctttaa ctgataatct gattacttta tttcgtgtgt
      721 ctatgatgat gatgatagtt acagaaccga cgaactagtc tgtacccgat caacaccgag
      781 acccgtggcg tcttcgacct caatggcgtc tggaacttca agctggacta cgggaaagga
      841 ctggaagaga agtggtacga aagcaagctg accgacacta ttagtatggc cgtcccaagc
      901 agttacaatg acattggcgt gaccaaggaa atccgcaacc atatcggata tgtctggtac
      961 gaacgtgagt tcacggtgcc ggcctatctg aaggatcagc gtatcgtgct ccgcttcggc
     1021 tctgcaactc acaaagcaat tgtctatgtc aatggtgagc tggtcgtgga gcacaagggc
     1081 ggattcctgc cattcgaagc ggaaatcaac aactcgctgc gtgatggcat gaatcgcgtc
     1141 accgtcgccg tggacaacat cctcgacgat agcaccctcc cggtggggct gtacagcgag
     1201 cgccacgaag agggcctcgg aaaagtcatt cgtaacaagc cgaacttcga cttcttcaac
     1261 tatgcaggcc tgcaccgtcc ggtgaaaatc tacacgaccc cgtttacgta cgtcgaggac
     1321 atctcggttg tgaccgactt caatggccca accgggactg tgacctatac ggtggacttt
     1381 caaggcaaag ccgagaccgt gaaagtgtcg gtcgtggatg aggaaggcaa agtggtcgca
     1441 agcaccgagg gcctgagcgg taacgtggag attccgaatg tcatcctctg ggaaccactg
     1501 aacacgtatc tctaccagat caaagtggaa ctggtgaacg acggactgac catcgatgtc
     1561 tatgaagagc cgttcggcgt gcggaccgtg gaagtcaacg acggcaagtt cctcatcaac
     1621 aacaaaccgt tctacttcaa gggctttggc aaacatgagg acactcctat caacggccgt
     1681 ggctttaacg aagcgagcaa tgtgatggat ttcaatatcc tcaaatggat cggcgccaac
     1741 agcttccgga ccgcacacta tccgtactct gaagagttga tgcgtcttgc ggatcgcgag
     1801 ggtctggtcg tgatcgacga gactccggca gttggcgtgc acctcaactt catggccacc
     1861 acgggactcg gcgaaggcag cgagcgcgtc agtacctggg agaagattcg gacgtttgag
     1921 caccatcaag acgttctccg tgaactggtg tctcgtgaca agaaccatcc aagcgtcgtg
     1981 atgtggagca tcgccaacga ggcggcgact gaggaagagg gcgcgtacga gtacttcaag
     2041 ccgttggtgg agctgaccaa ggaactcgac ccacagaagc gtccggtcac gatcgtgctg
     2101 tttgtgatgg ctaccccgga gacggacaaa gtcgccgaac tgattgacgt catcgcgctc
     2161 aatcgctata acggatggta cttcgatggc ggtgatctcg aagcggccaa agtccatctc
     2221 cgccaggaat ttcacgcgtg gaacaagcgt tgcccaggaa agccgatcat gatcactgag
     2281 tacggcgcag acaccgttgc gggctttcac gacattgatc cagtgatgtt caccgaggaa
     2341 tatcaagtcg agtactacca ggcgaaccac gtcgtgttcg atgagtttga gaacttcgtg
     2401 ggtgagcaag cgtggaactt cgcggacttc gcgacctctc agggcgtgat gcgcgtccaa
     2461 ggaaacaaga agggcgtgtt cactcgtgac cgcaagccga agctcgccgc gcacgtcttt
     2521 cgcgagcgct ggaccaacat tccagatttc ggctacaaga acgctagcca tcaccatcac
     2581 catcacgtgt gaattggtga ccagctcgaa tttccccgat cgttcaaaca tttggcaata
     2641 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     2701 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     2761 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     2821 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggggtacca
     2881 taacttcgta tagcatacat tatacgaagt tatgcatgca ttcgggtcaa ggcggaagcc
     2941 agcgcgccac cccacgtcag caaatacgga ggcgcggggt tgacggcgtc acccggtcct
     3001 aacggcgacc aacaaaccag ccagaagaaa ttacagtaaa aaaaaagtaa attgcacttt
     3061 gatccacctt ttattaccta agtctcaatt tggatcaccc ttaaacctat cttttcaatt
     3121 tgggccgggt tgtggtttgg actaccatga acaacttttc gtcatgtcta acttcccttt
     3181 cagcaaacat atgaaccata tatagaggag atcggccgta tactagagct gatgtgttta
     3241 aggtcgttga ttgcacgaga aaaaaaaatc caaatcgcaa caatagcaaa tttatctggt
     3301 tcaaagtgaa aagatatgtt taaaggtagt ccaaagtaaa acttatagat aataaaatgt
     3361 ggtccaaagc gtaattcact caaaaaaaat caacgagacg tgtaccaaac ggagacaaac
     3421 ggcatcttct cgaaatttcc caaccgctcg ctcgcccgcc tcgtcttccc ggaaaccgcg
     3481 gtggtttcag cgtggcggat tctccaagca gacggagacg tcacggcacg ggactcctcc
     3541 caccacccaa ccgccataaa taccagcccc ctcatctcct ctcctcgcat cagctccacc
     3601 cccgaaaaat ttctccccaa tctcgcgagg ctctcgtcgt cgaatcgaat cctctcgcgt
     3661 cctcaaggta cgctgcttct cctctcctcg cttcgtttcg attcgatttc ggacgggtga
     3721 ggttgttttg ttgctagatc cgattggtgg ttagggttgt cgatgtgatt atcgtgagat
     3781 gtttaggggt tgtagatctg atggttgtga tttgggcacg gttggttcga taggtggaat
     3841 cgtggttagg ttttgggatt ggatgttggt tctgatgatt ggggggaatt tttacggtta
     3901 gatgaattgt tggatgattc gattggggaa atcggtgtag atctgttggg gaattgtgga
     3961 actagtcatg cctgagtgat tggtgcgatt tgtagcgtgt tccatcttgt aggccttgtt
     4021 gcgagcatgt tcagatctac tgttccgctc ttgattgagt tattggtgcc ttgggttggt
     4081 gcaaacacag gctttaatat gttatatctg ttttgtgttt gatgtagatc tgtagggtag
     4141 ttcttcttag acatggttca attatgtagc ttgtgcgttt cgatttgatt tcatatgttc
     4201 acagattaga taatgatgaa ctcttttaat taattgtcaa tggtaaatag gaagtcttgt
     4261 cgctatatct gtcataatga tctcatgtta ctatctgcca gtaatttatg ctaagaacta
     4321 tattagaata tcatgttaca atctgtagta atatcatgtt acaatctgta gttcatctat
     4381 ataatctatt gtggtaattt ctttttacta tctgtgtgaa gattattgcc actagttcat
     4441 tctacttatt tctgaagttc aggatacgtg tgctgttact acctatctga atacatgtgt
     4501 gatgtgcctg ttactatctt tttgaataca tgtatgttct gttggaatat gtttgctgtt
     4561 tgatccgttg ttgtgtcctt aatcttgtgc tagttcttac cctatctgtt tggtgattat
     4621 ttcttgcaga ttcagatcgc tagcccacca tgaaaaagcc tgaactcacc gcgacgtctg
     4681 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     4741 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     4801 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     4861 cgctcccgat tccggaagtg cttgacattg gggagtttag cgagagcctg acctattgca
     4921 tctcccgccg ttcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     4981 ttctacaacc ggtcgcggag gctatggatg cgatcgctgc ggccgatctt agccagacga
     5041 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     5101 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     5161 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     5221 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     5281 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     5341 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     5401 ggaggcatcc ggagcttgca ggatcgccac gactccgggc gtatatgctc cgcattggtc
     5461 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     5521 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     5581 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     5641 gacgccccag cactcgtccg agggcaaaga aatagagtag atgccgaccg gatctgtcga
     5701 actagcataa cttcgtatag catacattat acgaagttat tcgacaagct cgagtttctc
     5761 cataataatg tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc
     5821 atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca
     5881 ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcc cccgaattaa
     5941 ttcggcgtta attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt
     6001 caatttgttt acaccacaat atatcctgcc accgttaacg ttacaccaca atatatcctg
     6061 ccaagatctc taattccggg gatcggaaat ccagaagccc gagaggttgc cgcctttcgg
     6121 gctttttctt tttcaaaaaa aaaaatttat aaaacgatct gttgcggccg gccgccgggt
     6181 tgtgggcaaa ggcgctggcg ctcgacggtg ggcaaccgct tgcggttgtc cacgggcgga
     6241 gccggtgcgc gtagcgcatt gtccacaagc caagggcgac caataattga tatatatatt
     6301 cataattgaa aagctaattg aacatactac ttgctgtaac tacttgccgg agcgaggggt
     6361 gtttgcaagc tgttgatctg aaagggctat tagcgttctc acgtgccttt ttgattagcg
     6421 atttcacgtg accttattag cgatttcacg tactccgatt agcgatttca cgtaccctga
     6481 ttagcgattt cacgtggata gtttttggag cgggccggaa agccccgtga atcaaggctt
     6541 tgcggggcat tagcggtttc acgtggataa ctaccctcta tccacaggct tccggggata
     6601 aaaaagcccg ctcgacggcg ggctgttgga tggggatcgc ctgaatcgcc ccatcatcca
     6661 gccagaaagt gagggagcca cggttgatga gagctttgtt gtaggtggac cagttggtga
     6721 ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat
     6781 ccttcaactc agcaaaagtt cgatttattc aacaaagcca cgttgtgtct caaaatctct
     6841 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     6901 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctcaaggc
     6961 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     7021 tcgggcaatc aggtgcgaca atctaccgat tgtatgggaa gcccgatgcg ccagagttgt
     7081 ttctgaaaca tggcaaaggt agcgttgcca atgttgttac agatgagatg gtcagactaa
     7141 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     7201 atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta ttagaagaat
     7261 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     7321 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     7381 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     7441 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     7501 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     7561 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     7621 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     7681 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     7741 cactagacca atgttacaca tatatacttt agattgattt aaaacttcat ttttaattta
     7801 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7981 gtttgccgga tcaagagcta ccaactcttc ttccgaaggt aactggcttc agcagagcgc
     8041 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     8101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     8161 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     8221 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     8281 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     8341 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     8401 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8461 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     8521 tacggttcct ggccttttgc tggccttttg ctcacatgag atctcaaaca aacacataca
     8581 gcgactta
//
