LOCUS       sequence-258397-        8195 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP mRNA from the Drosophila Hml promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8195)
  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 8195)
  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_pHml eGFP SV40
FEATURES             Location/Qualifiers
     source          1..8195
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             71..91
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    516..637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1387..1403
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1388..1404
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1422..1424,1425..1427,1428..2141)
                     /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(1422..1424,1425..1427,1428..2138)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1467..1488)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1728..1747)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2075..2096
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2150..2271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2981..2998)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3152..3740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3232..3251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3911..4702,4703..4771))
                     /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     4534..4553
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4772..4876)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4944..4962
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6358..6637
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             6683..7708
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6683..7708
                     /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          8071..8136
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
ORIGIN
        1 cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca
       61 ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt
      121 gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta
      181 caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac
      241 ctttataagt aggcataaca gttataatca taacatactg ttttttctta ctccacacag
      301 gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat
      361 ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca
      421 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
      481 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
      541 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
      601 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatt aattaaattt
      661 ggtacccaaa agttatttct gtaggcaggt tcaatacact ttactaattt tgacaccaat
      721 gcgccctcta gcgggctacg agacaagctg ctgctcgaac gcatgaaccg taaactgctt
      781 gcattcagta tttgcactgc ttgaaccgat aacttttaac ggtcacaatg tagtacgttg
      841 aatatttgaa attagttcac ccttggccaa cattaaagtg attatattcc accacccaag
      901 ttcaatgcca ttggagaagt ttctgcgtag gacgctttac atccagcagc tcttcgttga
      961 ccatataact tttgaatggc gacatatcag cagagggggc taccccaagt cagaaaattc
     1021 ttacacccat cgtggactct agtaaattat gacataggtc ccggaaatgc cgctatcagg
     1081 aaggcattag catcagctta aacttggcac agatttgtta cattcgcttt gctaatttaa
     1141 gaacggcgat gccgataagc tgatgatggt gctttttttg cggatcgtgg ataaatagtg
     1201 ggcttgagcc taacaattcc gagtgttctg ataaaccagc tacggaatcg ggcctataaa
     1261 aggggctccc cattcatggg gaacattagt cgcgatcgcg catcgaaatc gtcaagttca
     1321 gtggatataa ttacagtgaa aaagtgctaa gtgtaaaagt ttttaaaagt gagttctaag
     1381 ctggactcga ggtcgacggt atcgataagc ttgatatcac catggtgagc aagggcgagg
     1441 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1501 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1561 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1621 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1681 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1741 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1801 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1861 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1921 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1981 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2041 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2101 ccgccgggat cactctcggc atggacgagc tgtacaagta agcggccgca acttgtttat
     2161 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2221 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aactagtgga
     2281 tccactaggg gccgccgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
     2341 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
     2401 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac
     2461 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
     2521 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
     2581 gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag
     2641 aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa
     2701 catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg
     2761 gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
     2821 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
     2881 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
     2941 gtaaagtctg gaaacgcgga agtcagcgct cttccgcttc ctcgctcact gactcgctgc
     3001 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3061 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3121 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3181 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3241 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3301 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3361 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3421 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3481 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3541 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3601 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3661 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3721 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3781 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3841 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3901 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3961 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4021 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4081 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4141 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4201 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
     4261 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4321 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4381 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4441 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4501 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     4561 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4621 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4681 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4741 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4801 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4861 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4921 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat
     4981 tcacatttgt acgaattttt tttttatcaa aagttcgagt ttttcaccaa tttcctcatc
     5041 aaccgagcaa ggcaaacggc tttgaataat atggtgttat atatacatat atcaaatcgc
     5101 tgctgactgc gtgattgatg gccccaagat tacatattat cgaatcagga ttcagaagga
     5161 gatcaatgtc aaatgcggac aggaacatga aagacgcctg ttatgcgcaa ttaaaaattt
     5221 gggtttaatt gctgtggaaa ctgttgttgg cggcatctta agttcctgtt taacaacatc
     5281 aactacttat gtacgtagaa gcgtttaagc catttgcata cagatgagaa ctggcttttg
     5341 tgctaatcag tcaagatgac tccgatgatg atgactcatt acctgaccag ttttcgctgc
     5401 tttcttttca acaactactt gtatatgtat tgtatccaat agcaatacat tgaatttcca
     5461 tggtctagtc acgtattatc atttaattga caccaagtcg tgttattgtt gagctatcga
     5521 gttcagctca aacatttctt attcccatga ataagccggc aaaaatatgc aatctatgaa
     5581 agttaatata agcaaacctt actttgactc aataccaatg cactttgtgt cgataggttc
     5641 acgcaattga ggcgattatt ccgataaccc aagcgattga ctgttcccgt ttcgattcca
     5701 attgaaattt ggaaatgtac aatagttttg ctatatgctg tcaagtacgc tcttatcttc
     5761 tctgggtttt cttcagagtt tcgaaacgct tcttcttttt tttgtttttt tttttttgga
     5821 atctcgtatt ttggaagggg ctcccctctg gaatttgtta cactgtcgtt atcattgcga
     5881 acaagcggcc cgaagctatc agcgacttta acatttacaa tgcacttttt tacgaccaat
     5941 taaatgtaca ttttcctttc ttcgcccgtt gataagcgaa cgcgatgtgg cgcaggcaat
     6001 gtgttgctct tgcgacacaa acgcaatcaa aatggattca atttcgcttt ttcccagtga
     6061 aacgaagaac gaaccgacca tcatgatatg ctcctctgca tgttgcgtat tgaatcaatg
     6121 acaatttcaa ttaagccgcc cgttcgtcat gcgttttcgt gcgcttcgaa atgctgataa
     6181 cgctgctgtc ctccaactgc tttgcatgtg gacacaattc catttattta attcttttat
     6241 ttggatcggt taaattaaaa agcgccttgt tacgcattta acgttgtttc cggtgcgtgg
     6301 tggtttcatg cttctgggaa cggcaaatgg gtttaggatt gggaacccct catcatctgt
     6361 tggaatatac tattcaacct acaaaagtaa cgttaaacaa cactacttta tatttgatat
     6421 gaatggccac accttttatg ccataaaaca tattgtaaga gaataccact ctttttattc
     6481 cttctttcct tcttgtacgt tttttgctgt aagtaggtcg tggtgctggt gttgcagttg
     6541 aaataactta aaatataaat cataaaactc aaacataaac ttgactattt atttatttat
     6601 taagaaagga aatataaatt ataaattaca acaggttatg gacctgcagc caagcttggc
     6661 gctcgtccgg gggcaatgag atatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     6721 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     6781 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     6841 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     6901 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     6961 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     7021 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     7081 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     7141 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     7201 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     7261 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     7321 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     7381 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     7441 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     7501 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     7561 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     7621 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     7681 cagcactcgt ccgagggcaa aggaatagag tagatgccga ccgaacaaga gctgatttcg
     7741 agaacgcctc agccagcaac tcgcgcgagc ctagcaaggc aaatgcgaga gaacggcctt
     7801 acgcttggtg gcacagttct cgtccacagt tcgctaagct cgctcggctg ggtcgcggga
     7861 gggccggtcg cagtgattca ggcccttctg gattgtgttg gtccccaggg cacgattgtc
     7921 atgcccacgc actcgggtga tctgactgat cccgcagatt ggagatcgcc gcccgtgcct
     7981 gccgattggg tgcagatctt tgtgaaggaa ccttacttct gtggtgtgac ataattggac
     8041 aaactaccta cagagattta aagctctaag gtaaatataa aatttttaag tgtataatgt
     8101 gttaaactac tgattctaat tgtttgtgta ttttagattc caacctatgg aactgatgaa
     8161 tgggagcagt ggtggaatgc ctttaatgag gaaaa
//
