LOCUS       sequence-258485-        8171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP mRNA from the Drosophila CG8620 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8171)
  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 8171)
  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_pCG8620 eGFP SV40
FEATURES             Location/Qualifiers
     source          1..8171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    86..207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     933..949
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     934..950
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(968..970,971..973,974..1687)
                     /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(968..970,971..973,974..1684)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1013..1034)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1274..1293)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1621..1642
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1696..1817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2527..2544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2698..3286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2778..2797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3457..4248,4249..4317))
                     /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     4080..4099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4318..4422)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4490..4508
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5904..6183
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             6229..7254
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6229..7254
                     /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          7617..7682
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7812..7832
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
       61 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
      121 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
      181 tttgtccaaa ctcatcaatg tatcttatca tgtctggatt aattaaataa tgtgctaaac
      241 ctgatttatg caattattaa gatcccaaaa cattttcaaa aaattgtgta aatttcttaa
      301 tactttatat tttttaaaat tttcgatgtt ttggaaccac aaattaattg ttttaaaatt
      361 acacacttgg acgaaattca aaattaaaat tatatcagta gaatgtcata tttcataatt
      421 tcgtaaaata aaataaaaaa atatatgttt ataaaaatac gcagaatatt tatagtatat
      481 gtatatgcat actatccaga gatacaaaag taagaagaat ttctcaaaaa tatgaaatat
      541 gtttgaggtt acgtacggca atgaaattat gtaacattaa aaacacgtga tcaatggaag
      601 taatttccca aaaacccatg aatgcgattt ggttaaaaat agctcagatg acgtcacatt
      661 tatcgtgtgg tttgtatgag tatgtgactg ataaagaatg gcaaatgttt acagtccgaa
      721 agtcgcgatt agcttggcgc caaagttgcc tataaaagct catcgaccga atgccgcaga
      781 agccagtcaa attcaagtgt tcaagtgaag tagtcgcaac ataagcaaca gatcccgcag
      841 cagatatatc cagttcagag aaacagtagt taatcagtaa ttaaagtgtt gagtgaattt
      901 cttatcgggt cagttgtaaa ctagtgcaaa actcgaggtc gacggtatcg ataagcttga
      961 tatcaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1021 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1081 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1141 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1201 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1261 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1321 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1381 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1441 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1501 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1561 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1621 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1681 caagtaagcg gccgcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1741 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1801 ctcatcaatg tatcttaact agtggatcca ctaggggccg ccgacgcgag gctggatggc
     1861 cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat
     1921 gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct
     1981 taccagccta acttcgatca ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc
     2041 gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg tctgcctccc
     2101 cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac
     2161 ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg gagaactgtg
     2221 aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc
     2281 acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg
     2341 tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg
     2401 atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca
     2461 tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgctcttc
     2521 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2581 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3121 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3301 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3361 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3541 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3601 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3661 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3721 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat
     3781 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3841 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3901 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3961 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4021 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga
     4081 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4141 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4201 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4261 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4321 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4381 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4441 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4501 cacgaggccc tttcgtcttc aagaattcac atttgtacga attttttttt tatcaaaagt
     4561 tcgagttttt caccaatttc ctcatcaacc gagcaaggca aacggctttg aataatatgg
     4621 tgttatatat acatatatca aatcgctgct gactgcgtga ttgatggccc caagattaca
     4681 tattatcgaa tcaggattca gaaggagatc aatgtcaaat gcggacagga acatgaaaga
     4741 cgcctgttat gcgcaattaa aaatttgggt ttaattgctg tggaaactgt tgttggcggc
     4801 atcttaagtt cctgtttaac aacatcaact acttatgtac gtagaagcgt ttaagccatt
     4861 tgcatacaga tgagaactgg cttttgtgct aatcagtcaa gatgactccg atgatgatga
     4921 ctcattacct gaccagtttt cgctgctttc ttttcaacaa ctacttgtat atgtattgta
     4981 tccaatagca atacattgaa tttccatggt ctagtcacgt attatcattt aattgacacc
     5041 aagtcgtgtt attgttgagc tatcgagttc agctcaaaca tttcttattc ccatgaataa
     5101 gccggcaaaa atatgcaatc tatgaaagtt aatataagca aaccttactt tgactcaata
     5161 ccaatgcact ttgtgtcgat aggttcacgc aattgaggcg attattccga taacccaagc
     5221 gattgactgt tcccgtttcg attccaattg aaatttggaa atgtacaata gttttgctat
     5281 atgctgtcaa gtacgctctt atcttctctg ggttttcttc agagtttcga aacgcttctt
     5341 cttttttttg tttttttttt tttggaatct cgtattttgg aaggggctcc cctctggaat
     5401 ttgttacact gtcgttatca ttgcgaacaa gcggcccgaa gctatcagcg actttaacat
     5461 ttacaatgca cttttttacg accaattaaa tgtacatttt cctttcttcg cccgttgata
     5521 agcgaacgcg atgtggcgca ggcaatgtgt tgctcttgcg acacaaacgc aatcaaaatg
     5581 gattcaattt cgctttttcc cagtgaaacg aagaacgaac cgaccatcat gatatgctcc
     5641 tctgcatgtt gcgtattgaa tcaatgacaa tttcaattaa gccgcccgtt cgtcatgcgt
     5701 tttcgtgcgc ttcgaaatgc tgataacgct gctgtcctcc aactgctttg catgtggaca
     5761 caattccatt tatttaattc ttttatttgg atcggttaaa ttaaaaagcg ccttgttacg
     5821 catttaacgt tgtttccggt gcgtggtggt ttcatgcttc tgggaacggc aaatgggttt
     5881 aggattggga acccctcatc atctgttgga atatactatt caacctacaa aagtaacgtt
     5941 aaacaacact actttatatt tgatatgaat ggccacacct tttatgccat aaaacatatt
     6001 gtaagagaat accactcttt ttattccttc tttccttctt gtacgttttt tgctgtaagt
     6061 aggtcgtggt gctggtgttg cagttgaaat aacttaaaat ataaatcata aaactcaaac
     6121 ataaacttga ctatttattt atttattaag aaaggaaata taaattataa attacaacag
     6181 gttatggacc tgcagccaag cttggcgctc gtccgggggc aatgagatat gaaaaagcct
     6241 gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac
     6301 ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt
     6361 ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat
     6421 cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc
     6481 gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct
     6541 gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg
     6601 gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac
     6661 actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact
     6721 gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg
     6781 gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc
     6841 ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat
     6901 tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag
     6961 cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg
     7021 tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat
     7081 gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc
     7141 gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta
     7201 ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga atagagtaga
     7261 tgccgaccga acaagagctg atttcgagaa cgcctcagcc agcaactcgc gcgagcctag
     7321 caaggcaaat gcgagagaac ggccttacgc ttggtggcac agttctcgtc cacagttcgc
     7381 taagctcgct cggctgggtc gcgggagggc cggtcgcagt gattcaggcc cttctggatt
     7441 gtgttggtcc ccagggcacg attgtcatgc ccacgcactc gggtgatctg actgatcccg
     7501 cagattggag atcgccgccc gtgcctgccg attgggtgca gatctttgtg aaggaacctt
     7561 acttctgtgg tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa
     7621 atataaaatt tttaagtgta taatgtgtta aactactgat tctaattgtt tgtgtatttt
     7681 agattccaac ctatggaact gatgaatggg agcagtggtg gaatgccttt aatgaggaaa
     7741 acctgttttg ctcagaagaa atgccatcta gtgatgatga ggctactgct gactctcaac
     7801 attctactcc tccaaaaaag aagagaaagg tagaagaccc caaggacttt ccttcagaat
     7861 tgctaagttt tttgagtcat gctgtgttta gtaatagaac tcttgcttgc tttgctattt
     7921 acaccacaaa ggaaaaagct gcactgctat acaagaaaat tatggaaaaa tattctgtaa
     7981 cctttataag taggcataac agttataatc ataacatact gttttttctt actccacaca
     8041 ggcatagagt gtctgctatt aataactatg ctcaaaaatt gtgtaccttt agctttttaa
     8101 tttgtaaagg ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc
     8161 agccatacca c
//
