LOCUS       sequence-288591-        7207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of tubgenomic-PTC62-delIntron3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7207)
  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 7207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 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-PTC62-deltaIntron3_D
FEATURES             Location/Qualifiers
     source          1..7207
                     /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    3943..4064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3980..3999)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4034..4053
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4071..4526)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4208..4229)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4420..4439
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4553..4657
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4659..5016
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4659..4850
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4682..4702)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4867..5002
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5051..5845
                     /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             5051..5845
                     /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(5105..5124)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5715..5734
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6033..6052)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6077..6124
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6453..7041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6942..6961
                     /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 ccataagccg tgctggtgga tttggagcct ggtacaatgg actccgtgag gtctggtcca
     2701 tttggtcaga tcttcagacc agacaacttt gtgttcggcc agagtggtgc tggaaacaac
     2761 tgggccaagg gccactacac tgaaggagct gagctggttg attccgttct ggatgtggtc
     2821 cgaaaagagg ctgagagctg cgactgtctg cagggcttcc aactcactca ctcactgggt
     2881 ggaggtacag ggtctggtat gggcaccctc ctcattagca aaatccgcga ggagtatccc
     2941 gaccgcatca tgaacacctt cagcgtggtg ccctctccta aagtctcgga cactgtggtc
     3001 gagccctaca acgccacact gtccgtccat cagctagtag agaacacaga cgagacctat
     3061 tgtattgata acgaggccct gtacgatatc tgcttccgca cactcaaact cacaaccccc
     3121 acatacggag acctcaacca tctcgtctcc gccacaatga gcggtgtgac cacttgcttg
     3181 aggtttccag gccagttgaa cgctgatctc cgtaaattgg cggtcaacat ggtgcccttc
     3241 ccccgactgc acttcttcat gcctggcttc gcgcctctga ctagcagggg aagccagcag
     3301 tatcgtgcac ttaccgttcc cgaactcacc cagcagatgt tcgatgccaa aaacatgatg
     3361 gctgcctgcg acccacgtca cggccgttat ctgacggtcg ccgccgtctt ccgtggtcgc
     3421 atgtccatga aggaggtgga cgagcagatg ctcaacgtcc agaacaagaa cagcagctac
     3481 ttcgttgaat ggatcccaaa caacgtcaag accgccgtct gcgacattcc accacgaggc
     3541 ctcaagatgg ccgccacctt catcggcaac agcaccgcca tccaggagct cttcaagcgc
     3601 atctcagagc agttcacggc tatgttcagg cgcaaggctt tcctgcattg gtacaccgga
     3661 gagggcatgg atgagatgga gttcacagag gccgagagca acatgaacga cctggtgtcc
     3721 gagtaccagc agtaccagga cgccaccgct gaagaggaag gggagtttga ggaggaggga
     3781 gaggaggagc tcgcttaagg taccgcgggc ccgggatcca ccggatctag ataactgatc
     3841 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3901 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3961 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4021 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     4081 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4141 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4201 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4261 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4321 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4381 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4441 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4501 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4561 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4621 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     4681 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4741 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4801 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4861 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4921 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4981 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5041 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5101 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5161 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5221 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5281 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5341 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5401 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5461 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5521 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5581 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5641 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5701 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5761 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5821 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5881 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5941 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6001 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6061 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6121 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6181 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6241 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6301 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6361 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6421 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6481 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6541 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     6601 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6661 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6721 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6781 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6841 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6901 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6961 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7021 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7081 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7141 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
     7201 atatatg
//
