LOCUS       sequence-385678-        8207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP mRNA from the Drosophila RnrS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8207)
  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 8207)
  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_pRnrS eGFP SV40
FEATURES             Location/Qualifiers
     source          1..8207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1246..1311
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1441..1461
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1886..2007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2769..2785
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2770..2786
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(2804..2806,2807..2809,2810..3523)
                     /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(2804..2806,2807..2809,2810..3520)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2849..2870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3110..3129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3457..3478
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3532..3653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4363..4380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4534..5122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4614..4633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5293..6084,6085..6153))
                     /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     5916..5935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6154..6258)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6326..6344
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7740..8019
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             join(8065..8207,1..883)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             join(8065..8207,1..883)
                     /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"
ORIGIN
        1 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
       61 atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac
      121 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
      181 gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag
      241 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
      301 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
      361 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
      421 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
      481 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
      541 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
      601 cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg
      661 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
      721 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
      781 aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
      841 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagagtagat gccgaccgaa
      901 caagagctga tttcgagaac gcctcagcca gcaactcgcg cgagcctagc aaggcaaatg
      961 cgagagaacg gccttacgct tggtggcaca gttctcgtcc acagttcgct aagctcgctc
     1021 ggctgggtcg cgggagggcc ggtcgcagtg attcaggccc ttctggattg tgttggtccc
     1081 cagggcacga ttgtcatgcc cacgcactcg ggtgatctga ctgatcccgc agattggaga
     1141 tcgccgcccg tgcctgccga ttgggtgcag atctttgtga aggaacctta cttctgtggt
     1201 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
     1261 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
     1321 tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc
     1381 tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct
     1441 ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt
     1501 ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag
     1561 gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt
     1621 aggcataaca gttataatca taacatactg ttttttctta ctccacacag gcatagagtg
     1681 tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg
     1741 gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac
     1801 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     1861 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     1921 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     1981 tttgtccaaa ctcatcaatg tatcttatca tgtctggatt aattaacgat taaacgtact
     2041 tatcgataaa tatcaacagc cagctgttat gttattgtac tgttatttac atgattcgct
     2101 gtaatggaat tcattcaaag ttgttgaatt gagtatgtta gcacctgttg catttccaaa
     2161 taattcgaga tttccaaaaa aaaaattaat tttaagtcaa tgtccttgga aaaaaatttt
     2221 aactttgttt caccttgatt ttttatctaa ttgtagcaag aggaccaaat tttttcactc
     2281 aaaactaatt aagtggcgtt tcatcagctg ttttctggat tagtctaggg ttgtctcatt
     2341 gcatgaaata tcgatgataa aaaaatttca aaatttattt agtatttgaa actattaata
     2401 ttaatatttt tcaagtgaca agctgggaag ctaaacataa aattgtgcag taaggattcg
     2461 atttatggtt taagaaaaga aaactaccac cccataattg cattagattt accctaaatt
     2521 tataaaaagt gaattgacgc actcgacagc cctgattttc ccatagtttt cccatcacca
     2581 aaaatggcgg caaatcgaaa cagtttgccg cccggcataa aacccaatgt agctgtattt
     2641 cccagatcat ttgccacaca actttcaaac tgtacactta atacacgtcg tggtttaagt
     2701 gaattttacc agagaatcag agaagcgccc ctacctgcta ataataatcc attcaaaaac
     2761 atctcaactc gaggtcgacg gtatcgataa gcttgatatc accatggtga gcaagggcga
     2821 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2881 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2941 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     3001 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     3061 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     3121 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     3181 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     3241 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3301 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3361 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3421 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3481 cgccgccggg atcactctcg gcatggacga gctgtacaag taagcggccg caacttgttt
     3541 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3601 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaactagtg
     3661 gatccactag gggccgccga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     3721 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     3781 ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcattgg
     3841 accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg
     3901 gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     3961 ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
     4021 agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
     4081 aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt
     4141 gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     4201 cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     4261 tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     4321 tcgtaaagtc tggaaacgcg gaagtcagcg ctcttccgct tcctcgctca ctgactcgct
     4381 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4441 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4501 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4561 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4621 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4681 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4741 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4801 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4861 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4921 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     4981 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5041 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5101 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5161 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5221 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5401 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5581 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     5641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5881 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     5941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6241 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6301 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     6361 attcacattt gtacgaattt tttttttatc aaaagttcga gtttttcacc aatttcctca
     6421 tcaaccgagc aaggcaaacg gctttgaata atatggtgtt atatatacat atatcaaatc
     6481 gctgctgact gcgtgattga tggccccaag attacatatt atcgaatcag gattcagaag
     6541 gagatcaatg tcaaatgcgg acaggaacat gaaagacgcc tgttatgcgc aattaaaaat
     6601 ttgggtttaa ttgctgtgga aactgttgtt ggcggcatct taagttcctg tttaacaaca
     6661 tcaactactt atgtacgtag aagcgtttaa gccatttgca tacagatgag aactggcttt
     6721 tgtgctaatc agtcaagatg actccgatga tgatgactca ttacctgacc agttttcgct
     6781 gctttctttt caacaactac ttgtatatgt attgtatcca atagcaatac attgaatttc
     6841 catggtctag tcacgtatta tcatttaatt gacaccaagt cgtgttattg ttgagctatc
     6901 gagttcagct caaacatttc ttattcccat gaataagccg gcaaaaatat gcaatctatg
     6961 aaagttaata taagcaaacc ttactttgac tcaataccaa tgcactttgt gtcgataggt
     7021 tcacgcaatt gaggcgatta ttccgataac ccaagcgatt gactgttccc gtttcgattc
     7081 caattgaaat ttggaaatgt acaatagttt tgctatatgc tgtcaagtac gctcttatct
     7141 tctctgggtt ttcttcagag tttcgaaacg cttcttcttt tttttgtttt tttttttttg
     7201 gaatctcgta ttttggaagg ggctcccctc tggaatttgt tacactgtcg ttatcattgc
     7261 gaacaagcgg cccgaagcta tcagcgactt taacatttac aatgcacttt tttacgacca
     7321 attaaatgta cattttcctt tcttcgcccg ttgataagcg aacgcgatgt ggcgcaggca
     7381 atgtgttgct cttgcgacac aaacgcaatc aaaatggatt caatttcgct ttttcccagt
     7441 gaaacgaaga acgaaccgac catcatgata tgctcctctg catgttgcgt attgaatcaa
     7501 tgacaatttc aattaagccg cccgttcgtc atgcgttttc gtgcgcttcg aaatgctgat
     7561 aacgctgctg tcctccaact gctttgcatg tggacacaat tccatttatt taattctttt
     7621 atttggatcg gttaaattaa aaagcgcctt gttacgcatt taacgttgtt tccggtgcgt
     7681 ggtggtttca tgcttctggg aacggcaaat gggtttagga ttgggaaccc ctcatcatct
     7741 gttggaatat actattcaac ctacaaaagt aacgttaaac aacactactt tatatttgat
     7801 atgaatggcc acacctttta tgccataaaa catattgtaa gagaatacca ctctttttat
     7861 tccttctttc cttcttgtac gttttttgct gtaagtaggt cgtggtgctg gtgttgcagt
     7921 tgaaataact taaaatataa atcataaaac tcaaacataa acttgactat ttatttattt
     7981 attaagaaag gaaatataaa ttataaatta caacaggtta tggacctgca gccaagcttg
     8041 gcgctcgtcc gggggcaatg agatatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     8101 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     8161 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcct
//
