LOCUS       sequence-288592-        8224 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian Expression of tubgenomic-PTC90.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8224)
  AUTHORS   Wittkopp N, Huntzinger E, Weiler C, Sauliere J, Schmidt S, Sonawane 
            M, Izaurralde E
  TITLE     Nonsense-mediated mRNA decay effectors are essential for zebrafish 
            embryonic development and survival.
  JOURNAL   Mol Cell Biol. 2009 Jul . 29(13):3517-28.
  PUBMED    19414594
REFERENCE   2  (bases 1 to 8224)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol. 2009 Jul . 29(13):3517-28."
COMMENT     Sequence Label: pT7-EGFP-C1-tubgenomic-PTC90_D
FEATURES             Location/Qualifiers
     source          1..8224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        313..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     467..487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     561..580
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        561..579
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4960..5081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4997..5016)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5051..5070
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5088..5543)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5225..5246)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5437..5456
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5570..5674
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5676..6033
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5676..5867
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5699..5719)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5884..6019
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6068..6862
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6068..6862
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6122..6141)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6732..6751
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7050..7069)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7094..7141
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7470..8058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7959..7978
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
       61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      541 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 atgagggaga tcgtgcattt acaggctgga cagtgcggca accagattgg tgccaaggta
     1441 ggaaatgaac aaatttcatt ttttatcgta atcttgttaa gatttagatt tttgcgggaa
     1501 aagctgacgc taatccttgc ttgttttaga tccacactcg ggaaaattta aactggttaa
     1561 ttaatgttgt gtttgacttg actgttgtgt tagtcgtctt aatatttata ataaaaacat
     1621 gtttgactaa acgatatttg tctgaatatg atccttaatc tcacccgcaa caagttctta
     1681 aacgaatagc ctttgaagtc tattcctatc gaaggaaaat gattgggtta ctacaaaaat
     1741 gtcgattatt gtaagaaagt catcgtggtt tgaatgaact aatggtgaaa caatgcgtta
     1801 gttttagggt gttggccgcg cctgaccggc gtagagtctg attggcaagg gactcgactc
     1861 atctgagctt gtttggaaga gccgagtctc gttctacaat tgttgtcacc agactgaaag
     1921 gcatcgctga ctcatttgaa cggtttgctc tcgccagttc gtttgagtcg atttgcttga
     1981 acgaatcacg tgtcgtaatg tggccggttg tatctggttc agtgtgcatg gaaaagtgcg
     2041 ggaacgctaa tatcatgacg catgtttccc cctggcctca tacatggacc aaattcgact
     2101 catgtcgaac tattgagcga tatcttgggt taatattcga gacaaaagtg gaactgtgat
     2161 tcgtgcgatt tttctcttag ctcgggtggc gtaaaatggc cccggtgact tggcaatctc
     2221 cattttacaa gtgtggagcc cacataacgg gaagcgctgg tcattaaatg ttggattaac
     2281 attttttttt tacagaatct atcaccttaa tttctagatt cggttttctt tgtgggtctt
     2341 aagcattaac atgcatttat gttcctcttg aatccatctt atttcagttc tgggaagtca
     2401 ttagtgacga gcatggaatt gacccaacag gcagttacca tggcgacagt gaccttcagc
     2461 tggaccgaat taatgtttat tataatgaag ccacaggtga gagtctaatt actttactaa
     2521 tatttttcct tttttttatt tttttattta tttaattttt atgtattttt tttattaaat
     2581 ctttattgtg aaccctgaaa ttctggaatt tgtatttttt ctttaggtgg aaagtatgtt
     2641 ccacgtgccg tgctggtgga tttggagcct ggtacaatgg actccgtgag gtctggtcca
     2701 tttggtcaga tcttcagacc agacaactaa gtgttcggta tttaccttaa tttttcactc
     2761 ttgtcctcat tcagaatcaa gggttttccc tgcttaaaga attgaggttt tccaatgcca
     2821 ttaaattttg tgccatctct gcctattaac aaacggtcac gcctactgac agaccccaag
     2881 ttatgctcct gcactggtcg tgtgttttaa ctatattcag tagtagcctt tttaaagcct
     2941 tttttttttt tttttgtaag attacagtgt tttagctagc aaatttataa cttgttaacc
     3001 caaggaaaaa cccttagtaa aaacccttgt ttgtgctttg aggtaaacca aagtagttaa
     3061 cagtcgaaga gaatcttaca tttttatttt tgtacatttt tcaaaattgt actaggacta
     3121 gaatgggtgt tgaataattt taggaccaat ttggtgagcg gttatgatcc acttcaaatg
     3181 ttgaccgcta tagaacatct gatttgaata caatggatcc tgcagttgct ccaaacaaaa
     3241 tttgaatggc agattgtgac ctgatataga tctacagcaa tgatcacatt aaaaccatat
     3301 taactggatt ttttttattt tttcatatga agttttggca gtctattcat attaaaacga
     3361 tctcttagag acactttagg atattttaga tgagaaaata ccaatattta aaaattaaat
     3421 ccacatttga ttaaattatc ctaatttaaa acagatggaa aattatttat ttttaacggt
     3481 tgcattctca aaacaaaaaa aacgtacaaa atcaataatc tatcaaaccg tctatagaac
     3541 aatacgtgta atattttaca ctaattccta tccagagaaa caacaccagg ggcgtagcaa
     3601 ctgcaaggga acggtggaaa cgtccccctc acttttacag accggccatt tagaaaccga
     3661 ttattttaag aatgtaaact tttggatttc atacagctgc accccctgta tatcctattt
     3721 gcatgtatta actgaactcc tgaatttctt ataggccaga gtggtgctgg aaacaactgg
     3781 gccaagggcc actacactga aggagctgag ctggttgatt ccgttctgga tgtggtccga
     3841 aaagaggctg agagctgcga ctgtctgcag ggcttccaac tcactcactc actgggtgga
     3901 ggtacagggt ctggtatggg caccctcctc attagcaaaa tccgcgagga gtatcccgac
     3961 cgcatcatga acaccttcag cgtggtgccc tctcctaaag tctcggacac tgtggtcgag
     4021 ccctacaacg ccacactgtc cgtccatcag ctagtagaga acacagacga gacctattgt
     4081 attgataacg aggccctgta cgatatctgc ttccgcacac tcaaactcac aacccccaca
     4141 tacggagacc tcaaccatct cgtctccgcc acaatgagcg gtgtgaccac ttgcttgagg
     4201 tttccaggcc agttgaacgc tgatctccgt aaattggcgg tcaacatggt gcccttcccc
     4261 cgactgcact tcttcatgcc tggcttcgcg cctctgacta gcaggggaag ccagcagtat
     4321 cgtgcactta ccgttcccga actcacccag cagatgttcg atgccaaaaa catgatggct
     4381 gcctgcgacc cacgtcacgg ccgttatctg acggtcgccg ccgtcttccg tggtcgcatg
     4441 tccatgaagg aggtggacga gcagatgctc aacgtccaga acaagaacag cagctacttc
     4501 gttgaatgga tcccaaacaa cgtcaagacc gccgtctgcg acattccacc acgaggcctc
     4561 aagatggccg ccaccttcat cggcaacagc accgccatcc aggagctctt caagcgcatc
     4621 tcagagcagt tcacggctat gttcaggcgc aaggctttcc tgcattggta caccggagag
     4681 ggcatggatg agatggagtt cacagaggcc gagagcaaca tgaacgacct ggtgtccgag
     4741 taccagcagt accaggacgc caccgctgaa gaggaagggg agtttgagga ggagggagag
     4801 gaggagctcg cttaaggtac cgcgggcccg ggatccaccg gatctagata actgatcata
     4861 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4921 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4981 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5041 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
     5101 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     5161 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     5221 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     5281 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     5341 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     5401 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     5461 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     5521 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5581 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5641 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     5701 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5761 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5821 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5881 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5941 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6001 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     6061 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     6121 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     6181 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     6241 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     6301 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     6361 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     6421 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     6481 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     6541 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     6601 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     6661 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     6721 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6781 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6841 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6901 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6961 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     7021 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     7081 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     7141 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     7201 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     7261 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     7321 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7381 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7441 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7501 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7561 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     7621 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7681 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7741 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7801 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7861 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7921 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7981 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8041 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8101 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8161 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     8221 tatg
//
