LOCUS       sequence-258486-        8411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP mRNA from the Drosophila Sirup promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8411)
  AUTHORS   Tatomer DC, Elrod ND, Liang D, Xiao MS, Jiang JZ, Jonathan M, Huang 
            KL, Wagner EJ, Cherry S, Wilusz JE
  TITLE     The Integrator complex cleaves nascent mRNAs to attenuate 
            transcription.
  JOURNAL   Genes Dev. 2019 Sep 17. pii: gad.330167.119. doi: 
            10.1101/gad.330167.119.
  PUBMED    31530651
REFERENCE   2  (bases 1 to 8411)
  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: "Genes Dev. 
            2019 Sep 17. pii: gad.330167.119. doi: 10.1101/gad.330167.119."
COMMENT     Sequence Label: Hy_pSirup eGFP SV40
FEATURES             Location/Qualifiers
     source          1..8411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     455..471
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     456..472
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(490..492,493..495,496..1209)
                     /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(490..492,493..495,496..1206)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(535..556)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(796..815)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1143..1164
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1218..1339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2049..2066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2220..2808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2300..2319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2979..3770,3771..3839))
                     /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     3602..3621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3840..3944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4012..4030
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5426..5705
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             5751..6776
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5751..6776
                     /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          7139..7204
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7334..7354
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7779..7900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 atcttgagta aatggttggc ttttaatacg ttttagttgc aatatttttc gcagtttctt
       61 ttttacaaaa acggatttaa atttaaaagc cagcgttgcc agatagcatg tagctatagt
      121 tgccgttcaa tgcctagcag agtcatgcac cctggtgtgc gtctccgctg tgcgtatttt
      181 atggctatat ttagtacatt ttcagtgcca ccacaacata tggcacaata aataaagcaa
      241 ctaaacaagc aacgcgggct ataaaagcga aacgcagcct cgcagattgc aattcagacg
      301 cgaagcaaca agcaagtgac attgactccc aaccgaaagt gagacttttg agtaaacaca
      361 caaagtgtac ctgcgactaa gaagcgaaca aaaacgaaga ttcgccggca tcatcttgca
      421 taaccgataa aggaaatcca aacaaaaaac aaactcgagg tcgacggtat cgataagctt
      481 gatatcacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      541 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      601 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      661 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
      721 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      781 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
      841 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      901 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
      961 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1021 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1081 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1141 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1201 tacaagtaag cggccgcaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1261 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1321 aactcatcaa tgtatcttaa ctagtggatc cactaggggc cgccgacgcg aggctggatg
     1381 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     1441 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     1501 cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta tgccgcctcg
     1561 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc
     1621 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc
     1681 acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg cggagaactg
     1741 tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc
     1801 gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc
     1861 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     1921 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     1981 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgctct
     2041 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2641 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3121 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     3301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3361 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     3601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3841 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3961 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4021 atcacgaggc cctttcgtct tcaagaattc acatttgtac gaattttttt tttatcaaaa
     4081 gttcgagttt ttcaccaatt tcctcatcaa ccgagcaagg caaacggctt tgaataatat
     4141 ggtgttatat atacatatat caaatcgctg ctgactgcgt gattgatggc cccaagatta
     4201 catattatcg aatcaggatt cagaaggaga tcaatgtcaa atgcggacag gaacatgaaa
     4261 gacgcctgtt atgcgcaatt aaaaatttgg gtttaattgc tgtggaaact gttgttggcg
     4321 gcatcttaag ttcctgttta acaacatcaa ctacttatgt acgtagaagc gtttaagcca
     4381 tttgcataca gatgagaact ggcttttgtg ctaatcagtc aagatgactc cgatgatgat
     4441 gactcattac ctgaccagtt ttcgctgctt tcttttcaac aactacttgt atatgtattg
     4501 tatccaatag caatacattg aatttccatg gtctagtcac gtattatcat ttaattgaca
     4561 ccaagtcgtg ttattgttga gctatcgagt tcagctcaaa catttcttat tcccatgaat
     4621 aagccggcaa aaatatgcaa tctatgaaag ttaatataag caaaccttac tttgactcaa
     4681 taccaatgca ctttgtgtcg ataggttcac gcaattgagg cgattattcc gataacccaa
     4741 gcgattgact gttcccgttt cgattccaat tgaaatttgg aaatgtacaa tagttttgct
     4801 atatgctgtc aagtacgctc ttatcttctc tgggttttct tcagagtttc gaaacgcttc
     4861 ttcttttttt tgtttttttt tttttggaat ctcgtatttt ggaaggggct cccctctgga
     4921 atttgttaca ctgtcgttat cattgcgaac aagcggcccg aagctatcag cgactttaac
     4981 atttacaatg cactttttta cgaccaatta aatgtacatt ttcctttctt cgcccgttga
     5041 taagcgaacg cgatgtggcg caggcaatgt gttgctcttg cgacacaaac gcaatcaaaa
     5101 tggattcaat ttcgcttttt cccagtgaaa cgaagaacga accgaccatc atgatatgct
     5161 cctctgcatg ttgcgtattg aatcaatgac aatttcaatt aagccgcccg ttcgtcatgc
     5221 gttttcgtgc gcttcgaaat gctgataacg ctgctgtcct ccaactgctt tgcatgtgga
     5281 cacaattcca tttatttaat tcttttattt ggatcggtta aattaaaaag cgccttgtta
     5341 cgcatttaac gttgtttccg gtgcgtggtg gtttcatgct tctgggaacg gcaaatgggt
     5401 ttaggattgg gaacccctca tcatctgttg gaatatacta ttcaacctac aaaagtaacg
     5461 ttaaacaaca ctactttata tttgatatga atggccacac cttttatgcc ataaaacata
     5521 ttgtaagaga ataccactct ttttattcct tctttccttc ttgtacgttt tttgctgtaa
     5581 gtaggtcgtg gtgctggtgt tgcagttgaa ataacttaaa atataaatca taaaactcaa
     5641 acataaactt gactatttat ttatttatta agaaaggaaa tataaattat aaattacaac
     5701 aggttatgga cctgcagcca agcttggcgc tcgtccgggg gcaatgagat atgaaaaagc
     5761 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     5821 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     5881 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     5941 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     6001 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     6061 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     6121 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     6181 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     6241 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     6301 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     6361 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     6421 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     6481 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     6541 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     6601 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     6661 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     6721 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagagta
     6781 gatgccgacc gaacaagagc tgatttcgag aacgcctcag ccagcaactc gcgcgagcct
     6841 agcaaggcaa atgcgagaga acggccttac gcttggtggc acagttctcg tccacagttc
     6901 gctaagctcg ctcggctggg tcgcgggagg gccggtcgca gtgattcagg cccttctgga
     6961 ttgtgttggt ccccagggca cgattgtcat gcccacgcac tcgggtgatc tgactgatcc
     7021 cgcagattgg agatcgccgc ccgtgcctgc cgattgggtg cagatctttg tgaaggaacc
     7081 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt
     7141 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt
     7201 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga
     7261 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca
     7321 acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact ttccttcaga
     7381 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat
     7441 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatattctgt
     7501 aacctttata agtaggcata acagttataa tcataacata ctgttttttc ttactccaca
     7561 caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt
     7621 aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta gagatcataa
     7681 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     7741 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     7801 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     7861 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg attaattaaa
     7921 actccagaga atttcagtag ccctttacga tttcggcttc ttaacttatt tggttgataa
     7981 gccaagataa tgtccgggac ccaattccta atcgcctgct ctcaattacc gatgggactt
     8041 tgcgataaga acagggcgac tgggcgaccg tatagccgtg caactaacct ttgatataac
     8101 atgaaaaggg tgagctcgct ggatttcaac gatcgatggc tctaatcagc tttacatgtc
     8161 tttaatatga caacataatg aaaagcataa agaacattac aaactccaga taggcgcgat
     8221 tatacgtcgg tttcttatac aaaaaaaaaa aaaaatgata aaaatgccag ttgataatat
     8281 attaagaaat ggtttgaaca tttttgaatt taattattat gcagaagtga gcaaaaaaat
     8341 gtgcaatggc ttttaagaag tctgatagta attaaagcgt aataaaaaga aaatgcaact
     8401 aactgataaa t
//
