LOCUS       40924_22513        7698 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGPro4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7698)
  AUTHORS   Chen GQ, Thilmony R, Lin J-T.
  TITLE     Transformation of Lesquerella fendleri with the new binary vector 
            pGPro4-35S
  JOURNAL   Online J Biol Sci 11 (3), 90-95 (2012)
REFERENCE   2  (bases 1 to 7698)
  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   3  (bases 1 to 7698)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7698)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Online J 
            Biol Sci"; date: "2012"; volume: "11"; issue: "3"; pages: "90-95"
COMMENT     SGRef: number: 2; 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: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7698
                     /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"
     CDS             558..2363
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="LRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIA
                     VPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVM
                     EHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDF
                     FNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQV
                     VATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQFL
                     INHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWA
                     DEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDK
                     NHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     CDS             2388..3104
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3114..3131
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3163..3415
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3425..3458
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    3425..3458
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        3545..3724
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             3760..4782
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     protein_bind    complement(4817..4850)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4865..5039
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    complement(5117..5141)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(5151..5173)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     rep_origin      complement(5308..5743)
                     /direction=LEFT
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6034..6846
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7020..7608
                     /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 tggtagatct gactagttta cgtcctgtag aaaccccaac ccgtgaaatc aaaaaactcg
      601 acggcctgtg ggcattcagt ctggatcgcg aaaactgtgg aattgatcag cgttggtggg
      661 aaagcgcgtt acaagaaagc cgggcaattg ctgtgccagg cagttttaac gatcagttcg
      721 ccgatgcaga tattcgtaat tatgcgggca acgtctggta tcagcgcgaa gtctttatac
      781 cgaaaggttg ggcaggccag cgtatcgtgc tgcgtttcga tgcggtcact cattacggca
      841 aagtgtgggt caataatcag gaagtgatgg agcatcaggg cggctatacg ccatttgaag
      901 ccgatgtcac gccgtatgtt attgccggga aaagtgtacg tatcaccgtt tgtgtgaaca
      961 acgaactgaa ctggcagact atcccgccgg gaatggtgat taccgacgaa aacggcaaga
     1021 aaaagcagtc ttacttccat gatttcttta actatgccgg aatccatcgc agcgtaatgc
     1081 tctacaccac gccgaacacc tgggtggacg atatcaccgt ggtgacgcat gtcgcgcaag
     1141 actgtaacca cgcgtctgtt gactggcagg tggtggccaa tggtgatgtc agcgttgaac
     1201 tgcgtgatgc ggatcaacag gtggttgcaa ctggacaagg cactagcggg actttgcaag
     1261 tggtgaatcc gcacctctgg caaccgggtg aaggttatct ctatgaactg tgcgtcacag
     1321 ccaaaagcca gacagagtgt gatatctacc cgcttcgcgt cggcatccgg tcagtggcag
     1381 tgaagggcga acagttcctg attaaccaca aaccgttcta ctttactggc tttggtcgtc
     1441 atgaagatgc ggacttgcgt ggcaaaggat tcgataacgt gctgatggtg cacgaccacg
     1501 cattaatgga ctggattggg gccaactcct accgtacctc gcattaccct tacgctgaag
     1561 agatgctcga ctgggcagat gaacatggca tcgtggtgat tgatgaaact gctgctgtcg
     1621 gctttcagct gtctttaggc attggtttcg aagcgggcaa caagccgaaa gaactgtaca
     1681 gcgaagaggc agtcaacggg gaaactcagc aagcgcactt acaggcgatt aaagagctga
     1741 tagcgcgtga caaaaaccac ccaagcgtgg tgatgtggag tattgccaac gaaccggata
     1801 cccgtccgca aggtgcacgg gaatatttcg cgccactggc ggaagcaacg cgtaaactcg
     1861 acccgacgcg tccgatcacc tgcgtcaatg taatgttctg cgacgctcac accgatacca
     1921 tcagcgatct ctttgatgtg ctgtgcctga accgttatta cggatggtat gtccaaagcg
     1981 gcgatttgga aacggcagag aaggtactgg aaaaagaact tctggcctgg caggagaaac
     2041 tgcatcagcc gattatcatc accgaatacg gcgtggatac gttagccggg ctgcactcaa
     2101 tgtacaccga catgtggagt gaagagtatc agtgtgcatg gctggatatg tatcaccgcg
     2161 tctttgatcg cgtcagcgcc gtcgtcggtg aacaggtatg gaatttcgcc gattttgcga
     2221 cctcgcaagg catattgcgc gttggcggta acaagaaagg gatcttcact cgcgaccgca
     2281 aaccgaagtc ggcggctttt ctgctgcaaa aacgctggac tggcatgaac ttcggtgaaa
     2341 aaccgcagca gggaggcaaa caagctagcg gagcaggtgg cgctggtatg gtgagcaagg
     2401 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2461 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2521 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2581 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2641 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2701 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2761 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2821 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2881 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2941 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     3001 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3061 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagct agccaccacc
     3121 accaccacca cgtgtgaatt ggtgaccagc tcgaatttcc ccgatcgttc aaacatttgg
     3181 caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt
     3241 ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga
     3301 tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata
     3361 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact agatcggggg
     3421 taccataact tcgtatagca tacattatac gaagttatgc atgccgatac atgagaatta
     3481 agggagtcac gttatgaccc ccgccgatga cgcgggacaa gccgttttac gtttggaact
     3541 gacagaaccg caacgttgaa ggagccactc agccgcgggt ttctggagtt taatgagcta
     3601 agcacatacg tcagaaacca ttattgcgcg ttcaaaagtc gcctaaggtc actatcagct
     3661 agcaaatatt tcttgtcaaa aatgctccac tgacgttcca taaattcccc tcggtatcca
     3721 attagagtct catattcact ctcaactcac tagcccacca tgaaaaagcc tgaactcacc
     3781 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     3841 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     3901 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     3961 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggagtttag cgagagcctg
     4021 acctattgca tctcccgccg ttcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     4081 ctgcccgctg ttctacaacc ggtcgcggag gctatggatg cgatcgctgc ggccgatctt
     4141 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     4201 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     4261 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     4321 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     4381 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     4441 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     4501 tacttcgagc ggaggcatcc ggagcttgca ggatcgccac gactccgggc gtatatgctc
     4561 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     4621 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     4681 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     4741 agtggaaacc gacgccccag cactcgtccg agggcaaaga aatagagtag atgccgaccg
     4801 gatctgtcga actagcataa cttcgtatag catacattat acgaagttat tcgacaagct
     4861 cgagtttctc cataataatg tgtgagtagt tcccagataa gggaattagg gttcctatag
     4921 ggtttcgctc atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat
     4981 acttctatca ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcc
     5041 cccgaattaa ttcggcgtta attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt
     5101 gtctaagcgt caatttgttt acaccacaat atatcctgcc accgttaacg ttacaccaca
     5161 atatatcctg ccaagatctc taattccggg gatcggaaat ccagaagccc gagaggttgc
     5221 cgcctttcgg gctttttctt tttcaaaaaa aaaaatttat aaaacgatct gttgcggccg
     5281 gccgccgggt tgtgggcaaa ggcgctggcg ctcgacggtg ggcaaccgct tgcggttgtc
     5341 cacgggcgga gccggtgcgc gtagcgcatt gtccacaagc caagggcgac caataattga
     5401 tatatatatt cataattgaa aagctaattg aacatactac ttgctgtaac tacttgccgg
     5461 agcgaggggt gtttgcaagc tgttgatctg aaagggctat tagcgttctc acgtgccttt
     5521 ttgattagcg atttcacgtg accttattag cgatttcacg tactccgatt agcgatttca
     5581 cgtaccctga ttagcgattt cacgtggata gtttttggag cgggccggaa agccccgtga
     5641 atcaaggctt tgcggggcat tagcggtttc acgtggataa ctaccctcta tccacaggct
     5701 tccggggata aaaaagcccg ctcgacggcg ggctgttgga tggggatcgc ctgaatcgcc
     5761 ccatcatcca gccagaaagt gagggagcca cggttgatga gagctttgtt gtaggtggac
     5821 cagttggtga ttttgaactt ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc
     5881 gtgatctgat ccttcaactc agcaaaagtt cgatttattc aacaaagcca cgttgtgtct
     5941 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg
     6001 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
     6061 tgctcaaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     6121 cgcgataatg tcgggcaatc aggtgcgaca atctaccgat tgtatgggaa gcccgatgcg
     6181 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgttgttac agatgagatg
     6241 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     6301 actcctgatg atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta
     6361 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     6421 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     6481 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     6541 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     6601 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     6661 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     6721 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     6781 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     6841 tttttctaat cactagacca atgttacaca tatatacttt agattgattt aaaacttcat
     6901 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6961 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7021 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7081 gcggtggttt gtttgccgga tcaagagcta ccaactcttc ttccgaaggt aactggcttc
     7141 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7201 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7261 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7321 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7381 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7441 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7501 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7561 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7621 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgag atctcaaaca
     7681 aacacataca gcgactta
//
