LOCUS       sequence-258487-        8355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP mRNA from the Drosophila pst promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8355)
  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 8355)
  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_ppst eGFP SV40
FEATURES             Location/Qualifiers
     source          1..8355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          135..200
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             330..350
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    775..896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1806..1822
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1807..1823
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1841..1843,1844..1846,1847..2560)
                     /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(1841..1843,1844..1846,1847..2557)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1886..1907)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2147..2166)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2494..2515
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2569..2690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3400..3417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3571..4159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3651..3670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4330..5121,5122..5190))
                     /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     4953..4972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5191..5295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5363..5381
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6777..7056
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             7102..8127
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7102..8127
                     /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 gtgatctgac tgatcccgca gattggagat cgccgcccgt gcctgccgat tgggtgcaga
       61 tctttgtgaa ggaaccttac ttctgtggtg tgacataatt ggacaaacta cctacagaga
      121 tttaaagctc taaggtaaat ataaaatttt taagtgtata atgtgttaaa ctactgattc
      181 taattgtttg tgtattttag attccaacct atggaactga tgaatgggag cagtggtgga
      241 atgcctttaa tgaggaaaac ctgttttgct cagaagaaat gccatctagt gatgatgagg
      301 ctactgctga ctctcaacat tctactcctc caaaaaagaa gagaaaggta gaagacccca
      361 aggactttcc ttcagaattg ctaagttttt tgagtcatgc tgtgtttagt aatagaactc
      421 ttgcttgctt tgctatttac accacaaagg aaaaagctgc actgctatac aagaaaatta
      481 tggaaaaata ttctgtaacc tttataagta ggcataacag ttataatcat aacatactgt
      541 tttttcttac tccacacagg catagagtgt ctgctattaa taactatgct caaaaattgt
      601 gtacctttag ctttttaatt tgtaaagggg ttaataagga atatttgatg tatagtgcct
      661 tgactagaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
      721 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
      781 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      841 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
      901 gtctggatta attaactccg acagcgacta tatgtatttt gtattttgtg ttttgtctgg
      961 cactgccttt tggctccgtt ccgttccgtt cctttctctg ccgccgcttt cccctgcact
     1021 tttcctccgg cagatccagc tgatgttgct gtcgcgttgg agttatcgct acaatcgctg
     1081 cgccccgtgg attctcgatc gacacgcggc gatctgcgag attcgaatgg ggcaggagct
     1141 ggtgttgttg ctgctgctgc tgcaagatat cgcgagattg gcacgccgcg aacttaaaaa
     1201 actctgtatt tacaccgacg catgagactg acacgtaggg ttgtttggct cgtgctggaa
     1261 ccgataggcg tagctggagc ggagccggtt cgattgcacg gctatcgata ggcgggaggc
     1321 agcattcata gtttagggtt gccaaagctg gcagtactaa gtttagaatc tgatttttta
     1381 ggtcaccaag tactaaaaat gtttgcaagc aaaacttttg tcagcaagtt ttcaatgtaa
     1441 atgtatattt ttaaatggat ttgcctatat atatatatat atatatattt ttttttataa
     1501 tataatatgt gctcgctaga aatacaaagc aaatatctga ttagcatgag ctcatcgtct
     1561 cgttacattt tgggtttagc acgctagcag ctttttaaat tttgctcacc aaagaaggtg
     1621 acggtggaat acggggtatt catataaagg tatgatccct agagagcttt tctgccgtct
     1681 tgccttttcg gtatttcgag tacgatcgtt caattagtgt gcgccttaac ttaaaagctg
     1741 gtgattccgt ggcacaactc cattagcagc acaactgtgg caacggaaag gtgacaactt
     1801 agggctcgag gtcgacggta tcgataagct tgatatcacc atggtgagca agggcgagga
     1861 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1921 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1981 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2041 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2101 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2161 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2221 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2281 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2341 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2401 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2461 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2521 cgccgggatc actctcggca tggacgagct gtacaagtaa gcggccgcaa cttgtttatt
     2581 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2641 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta actagtggat
     2701 ccactagggg ccgccgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc
     2761 cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca
     2821 tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcattggacc
     2881 gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat
     2941 tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg
     3001 ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga
     3061 attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac
     3121 atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg
     3181 tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     3241 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     3301 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     3361 taaagtctgg aaacgcggaa gtcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     3421 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3481 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3541 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3601 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3661 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3721 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3781 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3841 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3901 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3961 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4021 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4081 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4141 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4201 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4261 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4321 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4381 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4441 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4501 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4561 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4621 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
     4681 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4741 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4801 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4861 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4921 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     4981 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5041 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5101 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5161 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5221 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5281 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     5341 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt
     5401 cacatttgta cgaatttttt ttttatcaaa agttcgagtt tttcaccaat ttcctcatca
     5461 accgagcaag gcaaacggct ttgaataata tggtgttata tatacatata tcaaatcgct
     5521 gctgactgcg tgattgatgg ccccaagatt acatattatc gaatcaggat tcagaaggag
     5581 atcaatgtca aatgcggaca ggaacatgaa agacgcctgt tatgcgcaat taaaaatttg
     5641 ggtttaattg ctgtggaaac tgttgttggc ggcatcttaa gttcctgttt aacaacatca
     5701 actacttatg tacgtagaag cgtttaagcc atttgcatac agatgagaac tggcttttgt
     5761 gctaatcagt caagatgact ccgatgatga tgactcatta cctgaccagt tttcgctgct
     5821 ttcttttcaa caactacttg tatatgtatt gtatccaata gcaatacatt gaatttccat
     5881 ggtctagtca cgtattatca tttaattgac accaagtcgt gttattgttg agctatcgag
     5941 ttcagctcaa acatttctta ttcccatgaa taagccggca aaaatatgca atctatgaaa
     6001 gttaatataa gcaaacctta ctttgactca ataccaatgc actttgtgtc gataggttca
     6061 cgcaattgag gcgattattc cgataaccca agcgattgac tgttcccgtt tcgattccaa
     6121 ttgaaatttg gaaatgtaca atagttttgc tatatgctgt caagtacgct cttatcttct
     6181 ctgggttttc ttcagagttt cgaaacgctt cttctttttt ttgttttttt ttttttggaa
     6241 tctcgtattt tggaaggggc tcccctctgg aatttgttac actgtcgtta tcattgcgaa
     6301 caagcggccc gaagctatca gcgactttaa catttacaat gcactttttt acgaccaatt
     6361 aaatgtacat tttcctttct tcgcccgttg ataagcgaac gcgatgtggc gcaggcaatg
     6421 tgttgctctt gcgacacaaa cgcaatcaaa atggattcaa tttcgctttt tcccagtgaa
     6481 acgaagaacg aaccgaccat catgatatgc tcctctgcat gttgcgtatt gaatcaatga
     6541 caatttcaat taagccgccc gttcgtcatg cgttttcgtg cgcttcgaaa tgctgataac
     6601 gctgctgtcc tccaactgct ttgcatgtgg acacaattcc atttatttaa ttcttttatt
     6661 tggatcggtt aaattaaaaa gcgccttgtt acgcatttaa cgttgtttcc ggtgcgtggt
     6721 ggtttcatgc ttctgggaac ggcaaatggg tttaggattg ggaacccctc atcatctgtt
     6781 ggaatatact attcaaccta caaaagtaac gttaaacaac actactttat atttgatatg
     6841 aatggccaca ccttttatgc cataaaacat attgtaagag aataccactc tttttattcc
     6901 ttctttcctt cttgtacgtt ttttgctgta agtaggtcgt ggtgctggtg ttgcagttga
     6961 aataacttaa aatataaatc ataaaactca aacataaact tgactattta tttatttatt
     7021 aagaaaggaa atataaatta taaattacaa caggttatgg acctgcagcc aagcttggcg
     7081 ctcgtccggg ggcaatgaga tatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     7141 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     7201 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     7261 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     7321 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     7381 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     7441 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     7501 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     7561 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     7621 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     7681 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     7741 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     7801 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     7861 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     7921 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     7981 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     8041 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     8101 agcactcgtc cgagggcaaa ggaatagagt agatgccgac cgaacaagag ctgatttcga
     8161 gaacgcctca gccagcaact cgcgcgagcc tagcaaggca aatgcgagag aacggcctta
     8221 cgcttggtgg cacagttctc gtccacagtt cgctaagctc gctcggctgg gtcgcgggag
     8281 ggccggtcgc agtgattcag gcccttctgg attgtgttgg tccccagggc acgattgtca
     8341 tgcccacgca ctcgg
//
