LOCUS       sequence-385677-        8079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP when there is readthrough transcription from the 
            Drosophila U5:34A gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8079)
  AUTHORS   Fujiwara R, Zhai SN, Liang D, Shah AP, Tracey M, Ma XK, Fields CJ, 
            Mendoza-Figueroa MS, Meline MC, Tatomer DC, Yang L, Wilusz JE
  TITLE     IntS6 and the Integrator phosphatase module tune the efficiency of 
            select premature transcription termination events.
  JOURNAL   Mol Cell. 2023 Nov 14:S1097-2765(23)00902-4. doi: 
            10.1016/j.molcel.2023.10.035.
  PUBMED    37995689
REFERENCE   2  (bases 1 to 8079)
  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. 
            2023 Nov 14:S1097-2765(23)00902-4. doi: 
            10.1016/j.molcel.2023.10.035."
COMMENT     Sequence Label: Hy_U5:34A eGFP SV40
FEATURES             Location/Qualifiers
     source          1..8079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    147..268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     902..918
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     903..919
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(937..939,940..942,943..1656)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(937..939,940..942,943..1653)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(982..1003)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1243..1262)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1590..1611
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1665..1786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2496..2513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2667..3255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2747..2766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3426..4217,4218..4286))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4049..4068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4287..4391)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4459..4477
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5873..6152
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             6198..7223
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6198..7223
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     intron          7586..7651
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7781..7801
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc agccatacca
       61 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
      121 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
      181 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
      241 gtttgtccaa actcatcaat gtatcttatc atgtctggat taattaaaac ttgaaacgga
      301 aaacgaaaag ggtctcgatg ggcaaacggc cagaaccatt tgggaattat aaatgtgaaa
      361 gtagaaagtt atcgtacttt ctattgcata tttaaggggt ttatggttca tatactacag
      421 acattcccgt tccaggtagc cacatgatat aatatgacta agatccctac cataaacata
      481 ctttaggggt tgtttgcgtt ttacaatcat tatgattccc aacatgttca agctcgttct
      541 aaatgatcgg gctgatactt ttcgttgtaa atttactctg gtttctcttc aattgtcgaa
      601 taaatctttc gccttttact aaagatttcc gtggagagga acactctaat gagtctaaaa
      661 taatcttttg tagtgcccgg cgacttcggt agctgggcca taaggaattc attgtaaagt
      721 tttttgctta ttattataag gtctggtggt catatacatg gactactcta ctaaggttac
      781 cagaagagtg atgaaatttt actcgaatcg agtttgtacc gtagacggcg ctaaatgtgc
      841 aataaatcag cttattatat tgtgcagtca attatatatt cttaatgtct atagtgactg
      901 ctcgaggtcg acggtatcga taagcttgat atcaccatgg tgagcaaggg cgaggagctg
      961 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     1021 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1081 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     1141 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     1201 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1261 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     1321 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     1381 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     1441 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     1501 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1561 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1621 gggatcactc tcggcatgga cgagctgtac aagtaagcgg ccgcaacttg tttattgcag
     1681 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1741 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacta gtggatccac
     1801 taggggccgc cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     1861 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     1921 ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcat tggaccgctg
     1981 atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta
     2041 ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc
     2101 acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg
     2161 gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
     2221 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct
     2281 ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
     2341 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
     2401 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
     2461 gtctggaaac gcggaagtca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2521 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     3121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3301 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3541 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3721 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4021 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     4081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4381 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     4441 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattcaca
     4501 tttgtacgaa tttttttttt atcaaaagtt cgagtttttc accaatttcc tcatcaaccg
     4561 agcaaggcaa acggctttga ataatatggt gttatatata catatatcaa atcgctgctg
     4621 actgcgtgat tgatggcccc aagattacat attatcgaat caggattcag aaggagatca
     4681 atgtcaaatg cggacaggaa catgaaagac gcctgttatg cgcaattaaa aatttgggtt
     4741 taattgctgt ggaaactgtt gttggcggca tcttaagttc ctgtttaaca acatcaacta
     4801 cttatgtacg tagaagcgtt taagccattt gcatacagat gagaactggc ttttgtgcta
     4861 atcagtcaag atgactccga tgatgatgac tcattacctg accagttttc gctgctttct
     4921 tttcaacaac tacttgtata tgtattgtat ccaatagcaa tacattgaat ttccatggtc
     4981 tagtcacgta ttatcattta attgacacca agtcgtgtta ttgttgagct atcgagttca
     5041 gctcaaacat ttcttattcc catgaataag ccggcaaaaa tatgcaatct atgaaagtta
     5101 atataagcaa accttacttt gactcaatac caatgcactt tgtgtcgata ggttcacgca
     5161 attgaggcga ttattccgat aacccaagcg attgactgtt cccgtttcga ttccaattga
     5221 aatttggaaa tgtacaatag ttttgctata tgctgtcaag tacgctctta tcttctctgg
     5281 gttttcttca gagtttcgaa acgcttcttc ttttttttgt tttttttttt ttggaatctc
     5341 gtattttgga aggggctccc ctctggaatt tgttacactg tcgttatcat tgcgaacaag
     5401 cggcccgaag ctatcagcga ctttaacatt tacaatgcac ttttttacga ccaattaaat
     5461 gtacattttc ctttcttcgc ccgttgataa gcgaacgcga tgtggcgcag gcaatgtgtt
     5521 gctcttgcga cacaaacgca atcaaaatgg attcaatttc gctttttccc agtgaaacga
     5581 agaacgaacc gaccatcatg atatgctcct ctgcatgttg cgtattgaat caatgacaat
     5641 ttcaattaag ccgcccgttc gtcatgcgtt ttcgtgcgct tcgaaatgct gataacgctg
     5701 ctgtcctcca actgctttgc atgtggacac aattccattt atttaattct tttatttgga
     5761 tcggttaaat taaaaagcgc cttgttacgc atttaacgtt gtttccggtg cgtggtggtt
     5821 tcatgcttct gggaacggca aatgggttta ggattgggaa cccctcatca tctgttggaa
     5881 tatactattc aacctacaaa agtaacgtta aacaacacta ctttatattt gatatgaatg
     5941 gccacacctt ttatgccata aaacatattg taagagaata ccactctttt tattccttct
     6001 ttccttcttg tacgtttttt gctgtaagta ggtcgtggtg ctggtgttgc agttgaaata
     6061 acttaaaata taaatcataa aactcaaaca taaacttgac tatttattta tttattaaga
     6121 aaggaaatat aaattataaa ttacaacagg ttatggacct gcagccaagc ttggcgctcg
     6181 tccgggggca atgagatatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
     6241 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
     6301 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
     6361 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     6421 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     6481 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     6541 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     6601 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     6661 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     6721 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     6781 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     6841 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     6901 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     6961 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     7021 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     7081 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     7141 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     7201 ctcgtccgag ggcaaaggaa tagagtagat gccgaccgaa caagagctga tttcgagaac
     7261 gcctcagcca gcaactcgcg cgagcctagc aaggcaaatg cgagagaacg gccttacgct
     7321 tggtggcaca gttctcgtcc acagttcgct aagctcgctc ggctgggtcg cgggagggcc
     7381 ggtcgcagtg attcaggccc ttctggattg tgttggtccc cagggcacga ttgtcatgcc
     7441 cacgcactcg ggtgatctga ctgatcccgc agattggaga tcgccgcccg tgcctgccga
     7501 ttgggtgcag atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact
     7561 acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa
     7621 actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga
     7681 gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag
     7741 tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt
     7801 agaagacccc aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag
     7861 taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata
     7921 caagaaaatt atggaaaaat attctgtaac ctttataagt aggcataaca gttataatca
     7981 taacatactg ttttttctta ctccacacag gcatagagtg tctgctatta ataactatgc
     8041 tcaaaaattg tgtaccttta gctttttaat ttgtaaagg
//
