LOCUS       Exported                7138 bp ds-DNA     circular SYN 01-AUG-2014
DEFINITION  2x35SpGREENLUC.
ACCESSION   .
VERSION     .
KEYWORDS    pGREENLUC2x23S
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7138)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 26, 2018 from SnapGene 4.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7138
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=pGREENLUC2x23S"
     primer_bind     5..21
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="/dnas_title=SK primer"
                     /note="/vntifkey=28"
     CDS             53..1705
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /note="/dnas_title=luciferase"
                     /note="/vntifkey=4"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(149..168)
                     /label=Primer 1
     polyA_signal    1762..1938
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
                     /note="/dnas_title=CaMV poly(A) signal"
                     /note="/vntifkey=25"
     misc_feature    1948..1972
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
                     /note="/dnas_title=RB T-DNA repeat"
                     /note="/vntifkey=21"
     rep_origin      complement(2063..2651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="/dnas_title=ori"
                     /note="/vntifkey=33"
     CDS             complement(2822..3637)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /note="/dnas_title=KanR"
                     /note="/vntifkey=4"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3928..4363
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
                     /note="/dnas_title=pSa ori"
                     /note="/vntifkey=33"
     misc_feature    4498..4520
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
                     /note="/dnas_title=LB T-DNA repeat"
                     /note="/vntifkey=21"
     promoter        4582..4927
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
                     /note="/dnas_title=CaMV 35S promoter"
                     /note="/vntifkey=30"
     CDS             4938..5873
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /note="/dnas_title=Rluc"
                     /note="/vntifkey=4"
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    5930..6106
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
                     /note="/dnas_title=CaMV poly(A) signal"
                     /note="/vntifkey=25"
     primer_bind     6270..6286
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="/dnas_title=M13 fwd"
                     /note="/vntifkey=28"
     promoter        6293..6311
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="/dnas_title=T7 promoter"
                     /note="/vntifkey=30"
     primer_bind     6337..6353
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="/dnas_title=KS primer"
                     /note="/vntifkey=28"
     promoter        6402..7128
                     /label=2X35S Promoter
                     /note="/dnas_title=2X35S Promoter"
                     /note="/vntifkey=29"
     TATA_signal     7096..7102
                     /label=TATA box
                     /note="/dnas_title=TATA box"
                     /note="/vntifkey=41"
ORIGIN
        1 gggggatcca ctagttctag agcggccgcc accgcggtgg agatcgaatt ccatggaaga
       61 cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg
      121 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
      181 agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt
      241 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
      301 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
      361 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc
      421 taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct
      481 cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
      541 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc
      601 agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg
      661 tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag
      721 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
      781 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
      841 cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca
      901 aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa
      961 atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt
     1021 cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac
     1081 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     1141 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     1201 cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa
     1261 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     1321 agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa
     1381 gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa
     1441 catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     1501 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     1561 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     1621 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     1681 gaagggcgga aagatcgccg tgtaattcta gagaattcgc tgaaatcacc agtctctctc
     1741 tacaaatcta tctctctcta ttttctccat aaataatgtg tgagtagttt cccgataagg
     1801 gaaattaggg ttcttatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat
     1861 gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc
     1921 agtactaaaa tccagatcca ctagccttga caggatatat tggcgggtaa actaagtcgc
     1981 tgtatgtgtt tgtttgagat ctcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2041 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2101 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2161 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2221 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2281 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2341 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2401 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2461 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2521 gcgctctgct gaagccagtt accttcggaa gaagagttgg tagctcttga tccggcaaac
     2581 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2641 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2701 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2761 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgtgtaacat tggtctagtg
     2821 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     2881 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     2941 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     3001 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     3061 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     3121 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     3181 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     3241 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     3301 attcttctaa tacctggaat gctgttttcc ctgggatcgc agtggtgagt aaccatgcat
     3361 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     3421 ttagtctgac catctcatct gtaacaacat tggcaacgct acctttgcca tgtttcagaa
     3481 acaactctgg cgcatcgggc ttcccataca atcggtagat tgtcgcacct gattgcccga
     3541 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     3601 gccttgagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     3661 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc
     3721 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg
     3781 atcagatcac gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa
     3841 tcaccaactg gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc
     3901 tggatgatgg ggcgattcag gcgatcccca tccaacagcc cgccgtcgag cgggcttttt
     3961 tatccccgga agcctgtgga tagagggtag ttatccacgt gaaaccgcta atgccccgca
     4021 aagccttgat tcacggggct ttccggcccg ctccaaaaac tatccacgtg aaatcgctaa
     4081 tcagggtacg tgaaatcgct aatcggagta cgtgaaatcg ctaataaggt cacgtgaaat
     4141 cgctaatcaa aaaggcacgt gagaacgcta atagcccttt cagatcaaca gcttgcaaac
     4201 acccctcgct ccggcaagta gttacagcaa gtagtatgtt caattagctt ttcaattatg
     4261 aatatatata tcaattattg gtcgcccttg gcttgtggac aatgcgctac gcgcaccggc
     4321 tccgcccgtg gacaaccgca agcggttgcc caccgtcgag cgccagcgcc tttgcccaca
     4381 acccggcggc cggccgcaac agatcgtttt ataaattttt ttttttgaaa aagaaaaagc
     4441 ccgaaaggcg gcaacctctc gggcttctgg atttccgatc cccggaatta gagatcttgg
     4501 caggatatat tgtggtgtaa cgttatcgta cccctactcc aaaaatgtca aagatacagt
     4561 ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatttcgg gaaacctcct
     4621 cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa aggaaggtgg
     4681 ctcctacaaa tgccatcatt gcgataaagg aaaggctatc attcaagatg cctctgccga
     4741 cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc
     4801 aaccacgtct tcaaagcaag tggattgatg tgacatctcc actgacgtaa gggatgacgc
     4861 acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat ttcatttgga
     4921 gaggacagcc caccaccatg acttcgaaag tttatgatcc agaacaaagg aaacggatga
     4981 taactggtcc gcagtggtgg gccagatgta aacaaatgaa tgttcttgat tcatttatta
     5041 attattatga ttcagaaaaa catgcagaaa atgctgttat ttttttacat ggtaacgcgg
     5101 cctcttctta tttatggcga catgttgtgc cacatattga gccagtagcg cggtgtatta
     5161 taccagacct tattggtatg ggcaaatcag gcaaatctgg taatggttct tataggttac
     5221 ttgatcatta caaatatctt actgcatggt ttgaacttct taatttacca aagaagatca
     5281 tttttgtcgg ccatgattgg ggtgcttgtt tggcatttca ttatagctat gagcatcaag
     5341 ataagatcaa agcaatagtt cacgctgaaa gtgtagtaga tgtgattgaa tcatgggatg
     5401 aatggcctga tattgaagaa gatattgcgt tgatcaaatc tgaagaagga gaaaaaatgg
     5461 ttttggagaa taacttcttc gtggaaacca tgttgccatc aaaaatcatg agaaagttag
     5521 aaccagaaga atttgcagca tatcttgaac cattcaaaga gaaaggtgaa gttcgtcgtc
     5581 caacattatc atggcctcgt gaaatcccgt tagtaaaagg tggtaaacct gacgttgtac
     5641 aaattgttag gaattataat gcttatctac gtgcaagtga tgatttacca aaaatgttta
     5701 ttgaatcgga cccaggattc ttttccaatg ctattgttga aggtgccaag aagtttccta
     5761 atactgaatt tgtcaaagta aaaggtcttc atttttcgca agaagatgca cctgatgaaa
     5821 tgggaaaata tatcaaatcg ttcgttgagc gagttctcaa aaatgaacaa taattctagc
     5881 cggtacgctg aaatcaccag tctctctcta caaatctatc tctctctatt ttctccataa
     5941 ataatgtgtg agtagtttcc cgataaggga aattagggtt cttatagggt ttcgctcatg
     6001 tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata
     6061 aaatttctaa ttcctaaaac caaaatccag tactaaaatc cagatcgata acattaacgt
     6121 ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6181 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     6241 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     6301 ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcgacggta tcgataagct
     6361 tgatatcgaa ttcctgcagg tcaacatggt ggagcacgac acacttgtct actccaaaaa
     6421 tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac aaagggtaat
     6481 atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg tgaagatagt
     6541 ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ccatcgttga
     6601 agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga
     6661 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata acatggtgga
     6721 gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc
     6781 aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc
     6841 tatctgtcac tttattgtga agatagtgga aaaggaaggt ggctcctaca aatgccatca
     6901 ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc gacagtggtc ccaaagatgg
     6961 acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca
     7021 agtggattga tgtgatatct ccactgacgt aagggatgac gcacaatccc actatccttc
     7081 gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggacct cgagaatt
//
