LOCUS       40924_22508        8419 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGPro4-35S, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8419)
  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 8419)
  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 8419)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8419)
  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..8419
                     /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"
     promoter        530..1206
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     CDS             1279..3084
                     /codon_start=1
                     /label=GUS
                     /note="beta-glucuronidase"
                     /translation="LRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIA
                     VPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVM
                     EHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDF
                     FNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQV
                     VATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQFL
                     INHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWA
                     DEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDK
                     NHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     CDS             3109..3825
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3835..3852
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3884..4136
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4146..4179
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    4146..4179
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        4266..4445
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             4481..5503
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     protein_bind    complement(5538..5571)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    5586..5760
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    complement(5838..5862)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(5872..5894)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA
                     (truncated)"
     rep_origin      complement(6029..6464)
                     /direction=LEFT
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6755..7567
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7741..8329
                     /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 cgggagcagc ttgccaacat ggtggagcac gacactctcg
      481 tctactccaa gaatatcaaa gatacagtct cagaagacca aagggctatt gagacttttc
      541 aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc tgtcacttca
      601 tcaaaaggac agtagaaaag gaaggtggca cctacaaatg ccatcattgc gataaaggaa
      661 aggctatcgt tcaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga
      721 ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
      781 aacatggtgg agcacgacac tcccgtctac tccaagaata tcaaagatac agtctcagaa
      841 gaccaaaggg ctattgagac ttttcaacaa agggtaatat cgggaaacct cctcggattc
      901 cattgcccag ctatctgtca cttcatcaaa aggacagtag aaaaggaagg tggcacctac
      961 aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctgc cgacagtggt
     1021 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg
     1081 tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc
     1141 cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggaca
     1201 cgctgaaatc accagtctct ctctacaaat ctatctctct cgagctttcg cagatccgcc
     1261 atggtagatc tgactagttt acgtcctgta gaaaccccaa cccgtgaaat caaaaaactc
     1321 gacggcctgt gggcattcag tctggatcgc gaaaactgtg gaattgatca gcgttggtgg
     1381 gaaagcgcgt tacaagaaag ccgggcaatt gctgtgccag gcagttttaa cgatcagttc
     1441 gccgatgcag atattcgtaa ttatgcgggc aacgtctggt atcagcgcga agtctttata
     1501 ccgaaaggtt gggcaggcca gcgtatcgtg ctgcgtttcg atgcggtcac tcattacggc
     1561 aaagtgtggg tcaataatca ggaagtgatg gagcatcagg gcggctatac gccatttgaa
     1621 gccgatgtca cgccgtatgt tattgccggg aaaagtgtac gtatcaccgt ttgtgtgaac
     1681 aacgaactga actggcagac tatcccgccg ggaatggtga ttaccgacga aaacggcaag
     1741 aaaaagcagt cttacttcca tgatttcttt aactatgccg gaatccatcg cagcgtaatg
     1801 ctctacacca cgccgaacac ctgggtggac gatatcaccg tggtgacgca tgtcgcgcaa
     1861 gactgtaacc acgcgtctgt tgactggcag gtggtggcca atggtgatgt cagcgttgaa
     1921 ctgcgtgatg cggatcaaca ggtggttgca actggacaag gcactagcgg gactttgcaa
     1981 gtggtgaatc cgcacctctg gcaaccgggt gaaggttatc tctatgaact gtgcgtcaca
     2041 gccaaaagcc agacagagtg tgatatctac ccgcttcgcg tcggcatccg gtcagtggca
     2101 gtgaagggcg aacagttcct gattaaccac aaaccgttct actttactgg ctttggtcgt
     2161 catgaagatg cggacttgcg tggcaaagga ttcgataacg tgctgatggt gcacgaccac
     2221 gcattaatgg actggattgg ggccaactcc taccgtacct cgcattaccc ttacgctgaa
     2281 gagatgctcg actgggcaga tgaacatggc atcgtggtga ttgatgaaac tgctgctgtc
     2341 ggctttcagc tgtctttagg cattggtttc gaagcgggca acaagccgaa agaactgtac
     2401 agcgaagagg cagtcaacgg ggaaactcag caagcgcact tacaggcgat taaagagctg
     2461 atagcgcgtg acaaaaacca cccaagcgtg gtgatgtgga gtattgccaa cgaaccggat
     2521 acccgtccgc aaggtgcacg ggaatatttc gcgccactgg cggaagcaac gcgtaaactc
     2581 gacccgacgc gtccgatcac ctgcgtcaat gtaatgttct gcgacgctca caccgatacc
     2641 atcagcgatc tctttgatgt gctgtgcctg aaccgttatt acggatggta tgtccaaagc
     2701 ggcgatttgg aaacggcaga gaaggtactg gaaaaagaac ttctggcctg gcaggagaaa
     2761 ctgcatcagc cgattatcat caccgaatac ggcgtggata cgttagccgg gctgcactca
     2821 atgtacaccg acatgtggag tgaagagtat cagtgtgcat ggctggatat gtatcaccgc
     2881 gtctttgatc gcgtcagcgc cgtcgtcggt gaacaggtat ggaatttcgc cgattttgcg
     2941 acctcgcaag gcatattgcg cgttggcggt aacaagaaag ggatcttcac tcgcgaccgc
     3001 aaaccgaagt cggcggcttt tctgctgcaa aaacgctgga ctggcatgaa cttcggtgaa
     3061 aaaccgcagc agggaggcaa acaagctagc ggagcaggtg gcgctggtat ggtgagcaag
     3121 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3181 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3241 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3301 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3361 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3421 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3481 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3541 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     3601 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     3661 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     3721 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3781 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaagc tagccaccac
     3841 caccaccacc acgtgtgaat tggtgaccag ctcgaatttc cccgatcgtt caaacatttg
     3901 gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt
     3961 tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag
     4021 atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat
     4081 atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcgggg
     4141 gtaccataac ttcgtatagc atacattata cgaagttatg catgccgata catgagaatt
     4201 aagggagtca cgttatgacc cccgccgatg acgcgggaca agccgtttta cgtttggaac
     4261 tgacagaacc gcaacgttga aggagccact cagccgcggg tttctggagt ttaatgagct
     4321 aagcacatac gtcagaaacc attattgcgc gttcaaaagt cgcctaaggt cactatcagc
     4381 tagcaaatat ttcttgtcaa aaatgctcca ctgacgttcc ataaattccc ctcggtatcc
     4441 aattagagtc tcatattcac tctcaactca ctagcccacc atgaaaaagc ctgaactcac
     4501 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     4561 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     4621 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     4681 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggagttta gcgagagcct
     4741 gacctattgc atctcccgcc gttcacaggg tgtcacgttg caagacctgc ctgaaaccga
     4801 actgcccgct gttctacaac cggtcgcgga ggctatggat gcgatcgctg cggccgatct
     4861 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     4921 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     4981 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     5041 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     5101 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     5161 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     5221 ctacttcgag cggaggcatc cggagcttgc aggatcgcca cgactccggg cgtatatgct
     5281 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     5341 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     5401 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     5461 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag aaatagagta gatgccgacc
     5521 ggatctgtcg aactagcata acttcgtata gcatacatta tacgaagtta ttcgacaagc
     5581 tcgagtttct ccataataat gtgtgagtag ttcccagata agggaattag ggttcctata
     5641 gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa
     5701 tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc
     5761 ccccgaatta attcggcgtt aattcagtac attaaaaacg tccgcaatgt gttattaagt
     5821 tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccgttaac gttacaccac
     5881 aatatatcct gccaagatct ctaattccgg ggatcggaaa tccagaagcc cgagaggttg
     5941 ccgcctttcg ggctttttct ttttcaaaaa aaaaaattta taaaacgatc tgttgcggcc
     6001 ggccgccggg ttgtgggcaa aggcgctggc gctcgacggt gggcaaccgc ttgcggttgt
     6061 ccacgggcgg agccggtgcg cgtagcgcat tgtccacaag ccaagggcga ccaataattg
     6121 atatatatat tcataattga aaagctaatt gaacatacta cttgctgtaa ctacttgccg
     6181 gagcgagggg tgtttgcaag ctgttgatct gaaagggcta ttagcgttct cacgtgcctt
     6241 tttgattagc gatttcacgt gaccttatta gcgatttcac gtactccgat tagcgatttc
     6301 acgtaccctg attagcgatt tcacgtggat agtttttgga gcgggccgga aagccccgtg
     6361 aatcaaggct ttgcggggca ttagcggttt cacgtggata actaccctct atccacaggc
     6421 ttccggggat aaaaaagccc gctcgacggc gggctgttgg atggggatcg cctgaatcgc
     6481 cccatcatcc agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga
     6541 ccagttggtg attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg
     6601 cgtgatctga tccttcaact cagcaaaagt tcgatttatt caacaaagcc acgttgtgtc
     6661 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     6721 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     6781 ttgctcaagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     6841 tcgcgataat gtcgggcaat caggtgcgac aatctaccga ttgtatggga agcccgatgc
     6901 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgttgtta cagatgagat
     6961 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     7021 tactcctgat gatgcatggt tactcaccac tgcgatccca gggaaaacag cattccaggt
     7081 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     7141 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     7201 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     7261 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     7321 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     7381 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     7441 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     7501 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     7561 gtttttctaa tcactagacc aatgttacac atatatactt tagattgatt taaaacttca
     7621 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7681 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7741 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7801 agcggtggtt tgtttgccgg atcaagagct accaactctt cttccgaagg taactggctt
     7861 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7921 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7981 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     8041 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     8101 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     8161 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     8221 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     8281 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     8341 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatga gatctcaaac
     8401 aaacacatac agcgactta
//
