LOCUS       sequence-358321-        8201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of tubgenomic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8201)
  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 8201)
  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_D
FEATURES             Location/Qualifiers
     source          1..8201
                     /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    4937..5058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4974..4993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5028..5047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5065..5520)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5202..5223)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5414..5433
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5547..5651
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5653..6010
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5653..5844
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5676..5696)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5861..5996
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6045..6839
                     /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             6045..6839
                     /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(6099..6118)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6709..6728
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7027..7046)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7071..7118
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7447..8035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7936..7955
                     /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 gatttaggtt tttgcgggaa
     1501 aagctgacac taatccttgc ttgttttaga tccacactcg ggaaaattta aactggctaa
     1561 ttaatgttgt gtttgacttg actgttgcgt tagtcgtctt aatatttgta ataaaaacat
     1621 gtttgactaa acgtcatttg tctgaatatg atccttaatc tcacccgcaa caagttctta
     1681 aacgaatagc ctttgaagtc tattcctatc gaaggaaaat gattgggtta ctacaaaaat
     1741 gtcgattatt gtaagaaagt catcgtggtt tgaatgaact aatggtgaaa caatgcgtta
     1801 gttttagggt gttggccgcg cctgaccggc gtagagtctg attcgcaagg gactcgactc
     1861 acctgagctt gtttggaaga gccgagtctc gttctacaat tgttgtcacc agactgaaag
     1921 gcaccgctga ctcatttgaa cggtttgctc tcgccagttc gtttgagtcg atttgcttga
     1981 acgaatcacg tgtcgtaatg tggccggttg tatctggttc agtgtgtatg gaaaagtgcg
     2041 ggaacgctaa tatcatgacg catgtttccc cctggcctct tgcatggacc aaattcgact
     2101 catgtcgaaa ctttgagcga tatcttgggt taatattcga gacaatagtg gaactgtgat
     2161 tcgtgcgatt tttctctaag 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 ttttttaaat ttttttattt atttattttt taatgtattt taattaaatc
     2581 tttattgtga accctgaaat tctggaattt gtatttttct ttaggtggaa agtatgttcc
     2641 acgtgccgtg ctggtggatt tggagcctgg tacaatggac tccgtgaggt ctggtccatt
     2701 tggtcagatc ttcggaccag acaactttgt gttcggtatt taccttaatt tttcactctt
     2761 gccctcattc agaatccagg gttttccctg cttaaagaat tgaggttttc caatgccatt
     2821 aaattttgtg ccatctctgc ctattaacag tcacgcctac tgacagaccc caagttatgc
     2881 tcctgcactg gtcgtgtgtt ttaactatat tcagtagtag cctctttaaa caatattttt
     2941 tttgtaagat tacagtgttt tagctagcaa atttataact tgttaaccca aggaaaaacc
     3001 cttagtaaac ctcgtttgtg cttttatttg aggtaaacta aagtagttga cagtcgaaca
     3061 gaatcttaaa ttttattttt gtacattttt caaaattgta ctaggactag aatggatgtt
     3121 gaataatttt agaaccaatt tggtgagcgg ttatgatcca cttcaaatgt tgactgctat
     3181 aaaacatctg atttgaatac aatggatcct gcagttgctc cgaacaaaat ttgaatggca
     3241 gattgtgacc tgatatagat ctacagcaat aatcgcatta aaaccatatt aactggattt
     3301 tttttcatat gaagttttgg cagtctattc atattaaaac gatcacttgg agacacttta
     3361 ggatatttta gatgagaaaa taccaatatt taaaaattaa atccacattt gattaaatta
     3421 tcctaattta aaacagatgg aaaattattt atttttaacg gttgcattct caaaacaaaa
     3481 aacgtacaaa atcaatctat caaacagtct atagaacaat acgtgtaata ttttacacta
     3541 attcctatcc agagaaacaa caccaggggc gtagcaactg caagggaacg gtggaaacgt
     3601 ccccctcact tttacagacc ggccatttag aaaccgatta ttttaagaat gtaaactttt
     3661 ggatttcata cagctgcacc ctccgtatat cctatttgca tgtattaact gaactcctga
     3721 atttcttata ggccaaagtg gtgctggaaa caactgggcc aagggccact acactgaagg
     3781 agctgagctg gttgattccg ttctggatgt ggtccgaaaa gaggctgaga gctgcgactg
     3841 tctgcagggc ttccagctca ctcactcact gggtggaggt acagggtctg gtatgggcac
     3901 cctcctcatt agcaaaatcc gcgaggagta tcccgaccgc atcatgaaca ccttcagcgt
     3961 ggtgccctct cctaaagtct cggacactgt ggtcgagccc tacaacgcca cactgtccgt
     4021 ccatcagcta gtagagaaca cagacgagac ctattgtatt gataacgagg ccctgtacga
     4081 tatctgcttc cgcacactca aactcacaac ccccacatac ggagacctca accatctcgt
     4141 ctccgccaca atgagcggtg tgaccacttg cttgaggttt ccaggccagt tgaacgctga
     4201 tctccgtaaa ttggcggtca acatggtgcc cttcccccga ctgcacttct tcatgcctgg
     4261 cttcgcgcct ctgactagca ggggaagcca gcagtatcgt gcacttaccg ttcccgaact
     4321 cacccagcag atgttcgatg ccaaaaacat gatggctgcc tgcgacccac gtcacggccg
     4381 ttatctgacg gtcgccgccg tcttccgagg tcgcatgtcc atgaaggagg tggacgagca
     4441 gatgctcaac gtccagaaca agaacagcag ctacttcgtt gaatggatcc caaacaacgt
     4501 caagaccgcc gtctgcgaca ttccaccacg aggcctcaag atggccgcca ccttcatcgg
     4561 caacagcact gccatccagg agctcttcaa gcgcatctca gagcagttca cggctatgtt
     4621 caggcgcaag gctttcctgc attggtacac cggagagggc atggatgaga tggagttcac
     4681 agaggccgag agcaacatga acgacctggt gtccgagtac cagcagtacc aggacgccac
     4741 cgctgaagag gaaggggagt ttgaggagga gggagaggag gagctcgctt aaggtaccgc
     4801 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     4861 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4921 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4981 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5041 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     5101 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5161 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5221 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5281 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5341 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5401 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5461 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5521 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5581 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5641 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     5701 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5761 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5821 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5881 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5941 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6001 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     6061 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     6121 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     6181 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     6241 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     6301 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     6361 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     6421 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     6481 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6541 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     6601 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6661 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6721 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6781 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6841 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6901 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6961 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     7021 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     7081 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     7141 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     7201 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     7261 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     7321 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     7381 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     7441 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     7501 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7561 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     7621 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7681 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7741 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7801 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7861 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7921 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7981 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8041 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8101 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat
     8161 agtaatcaat tacggggtca ttagttcata gcccatatat g
//
