LOCUS       sequence-258488-        8886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP mRNA from the Drosophila CG6770 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8886)
  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 8886)
  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_pCG6770 eGFP SV40
FEATURES             Location/Qualifiers
     source          1..8886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    223..344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1785..1801
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1786..1802
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1820..1822,1823..1825,1826..2539)
                     /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(1820..1822,1823..1825,1826..2536)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1865..1886)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2126..2145)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2473..2494
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2548..2669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3379..3396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3550..4138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3630..3649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4309..5100,5101..5169))
                     /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     4932..4951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5170..5274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5342..5360
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6756..7035
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             7081..8106
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7081..8106
                     /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          8469..8534
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8664..8684
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cacacaggca tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct
       61 ttttaatttg taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc
      121 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
      181 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
      241 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
      301 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggattaat
      361 taagaataaa acggaaggtt accccattca taacttctca gagaaagaga tttgtgaggc
      421 aaaggtcaat agtaaatact tagaataact atgttgagaa gagactgtat cagcaagtta
      481 agaacatttg ctatatttac aaaacgagac gtgggatctc tcgttctgag ttctagtaca
      541 acaaaattgt atctagaata ttcacggaaa cccagtttct ttttaaagcc attaaaaaag
      601 aaccatcgag aaatctaaag attgatttgc taccaacgaa ttgtaaataa agtggtatat
      661 tgtggcaggt tcttgaaaca acattgttga ggccgttcta tcttatctaa gatggtgact
      721 acgatctatt gcgtcgtttg cagacgaact ttcagaacga atacctgcca aatgactaac
      781 tagagcgata gttagcacct tatcaacgcc agaaggctat gtatatatgt atgtacgtaa
      841 catttgttta tataaaatcc attaaaccaa aacaaactga aagcttttcg ctattcagaa
      901 ataaacaaat gcgttgcctg gtgggaggga agtgggtcaa ccgatcaggt tctaatgcac
      961 caaaataaat atccattatg agtatatctt ttgtaaatca aaaatgttat tcgaattata
     1021 attattgcta aattgtttct tgtgccctaa cattcaaata ttgttgagtg taaaaaaatt
     1081 aacaaatttt gagcagtgta tccagaacat atgatgcatt taattctata tttattgggt
     1141 ggagaaacaa atgcagggta aatgcacttt gatggtcgtt tgcggagggc caaggtgggt
     1201 gtgtgttatg taaccactca ggaacgtgtg tatgcttaaa agttcttaaa ttattaaaat
     1261 aaatgaacaa ggcgaaataa tatgtttggc gatttcagtt tttttgttta ccgcgcactt
     1321 tgtaaacagc aaaaaattaa attgcgcagc aaaaattgtc aaggaggcac tctcttgagc
     1381 gcaaaagggt tgtcaggccc aataactatg aattttaata actaaaaatt cttttacata
     1441 aaaaagagca aatatatttt gtaaaagttc agtagaaagt tatagaaaaa ttaaatattt
     1501 ttagtaacta cacgtaagca acaattaaat gcccttctag tagtttcttt agctaaatcc
     1561 ggcaactctg acggagagag agctttggtt acggaacaga aagcttgcgc gcctataaaa
     1621 gccgctgctc gaccgaaaaa aattcaattc cgttcgcagc tcagcgaaaa gtgaacaaaa
     1681 aaagcagaaa agaaatttgg aaagcagcct gcaagtttgc agagaaacaa aaagcaaaca
     1741 ccaaccaaga acgcagttag ttttgcaacc ttaaatcgcc attctcgagg tcgacggtat
     1801 cgataagctt gatatcacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1861 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1921 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1981 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2041 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2101 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2161 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2221 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2281 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2341 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2401 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2461 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2521 ggacgagctg tacaagtaag cggccgcaac ttgtttattg cagcttataa tggttacaaa
     2581 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2641 ggtttgtcca aactcatcaa tgtatcttaa ctagtggatc cactaggggc cgccgacgcg
     2701 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     2761 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     2821 gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta
     2881 tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct
     2941 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga
     3001 agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg
     3061 cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc
     3121 tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg
     3181 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
     3241 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
     3301 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     3361 tcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3421 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3481 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3541 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3601 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3661 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3721 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3781 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3841 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3901 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3961 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4021 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4081 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4201 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4261 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4381 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4441 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4501 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4621 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4741 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4801 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4921 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4981 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5161 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5221 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5281 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     5341 aaataggcgt atcacgaggc cctttcgtct tcaagaattc acatttgtac gaattttttt
     5401 tttatcaaaa gttcgagttt ttcaccaatt tcctcatcaa ccgagcaagg caaacggctt
     5461 tgaataatat ggtgttatat atacatatat caaatcgctg ctgactgcgt gattgatggc
     5521 cccaagatta catattatcg aatcaggatt cagaaggaga tcaatgtcaa atgcggacag
     5581 gaacatgaaa gacgcctgtt atgcgcaatt aaaaatttgg gtttaattgc tgtggaaact
     5641 gttgttggcg gcatcttaag ttcctgttta acaacatcaa ctacttatgt acgtagaagc
     5701 gtttaagcca tttgcataca gatgagaact ggcttttgtg ctaatcagtc aagatgactc
     5761 cgatgatgat gactcattac ctgaccagtt ttcgctgctt tcttttcaac aactacttgt
     5821 atatgtattg tatccaatag caatacattg aatttccatg gtctagtcac gtattatcat
     5881 ttaattgaca ccaagtcgtg ttattgttga gctatcgagt tcagctcaaa catttcttat
     5941 tcccatgaat aagccggcaa aaatatgcaa tctatgaaag ttaatataag caaaccttac
     6001 tttgactcaa taccaatgca ctttgtgtcg ataggttcac gcaattgagg cgattattcc
     6061 gataacccaa gcgattgact gttcccgttt cgattccaat tgaaatttgg aaatgtacaa
     6121 tagttttgct atatgctgtc aagtacgctc ttatcttctc tgggttttct tcagagtttc
     6181 gaaacgcttc ttcttttttt tgtttttttt tttttggaat ctcgtatttt ggaaggggct
     6241 cccctctgga atttgttaca ctgtcgttat cattgcgaac aagcggcccg aagctatcag
     6301 cgactttaac atttacaatg cactttttta cgaccaatta aatgtacatt ttcctttctt
     6361 cgcccgttga taagcgaacg cgatgtggcg caggcaatgt gttgctcttg cgacacaaac
     6421 gcaatcaaaa tggattcaat ttcgcttttt cccagtgaaa cgaagaacga accgaccatc
     6481 atgatatgct cctctgcatg ttgcgtattg aatcaatgac aatttcaatt aagccgcccg
     6541 ttcgtcatgc gttttcgtgc gcttcgaaat gctgataacg ctgctgtcct ccaactgctt
     6601 tgcatgtgga cacaattcca tttatttaat tcttttattt ggatcggtta aattaaaaag
     6661 cgccttgtta cgcatttaac gttgtttccg gtgcgtggtg gtttcatgct tctgggaacg
     6721 gcaaatgggt ttaggattgg gaacccctca tcatctgttg gaatatacta ttcaacctac
     6781 aaaagtaacg ttaaacaaca ctactttata tttgatatga atggccacac cttttatgcc
     6841 ataaaacata ttgtaagaga ataccactct ttttattcct tctttccttc ttgtacgttt
     6901 tttgctgtaa gtaggtcgtg gtgctggtgt tgcagttgaa ataacttaaa atataaatca
     6961 taaaactcaa acataaactt gactatttat ttatttatta agaaaggaaa tataaattat
     7021 aaattacaac aggttatgga cctgcagcca agcttggcgc tcgtccgggg gcaatgagat
     7081 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     7141 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     7201 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     7261 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     7321 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     7381 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     7441 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     7501 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     7561 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     7621 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     7681 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     7741 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     7801 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     7861 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     7921 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     7981 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     8041 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     8101 gaatagagta gatgccgacc gaacaagagc tgatttcgag aacgcctcag ccagcaactc
     8161 gcgcgagcct agcaaggcaa atgcgagaga acggccttac gcttggtggc acagttctcg
     8221 tccacagttc gctaagctcg ctcggctggg tcgcgggagg gccggtcgca gtgattcagg
     8281 cccttctgga ttgtgttggt ccccagggca cgattgtcat gcccacgcac tcgggtgatc
     8341 tgactgatcc cgcagattgg agatcgccgc ccgtgcctgc cgattgggtg cagatctttg
     8401 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
     8461 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg
     8521 tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct
     8581 ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg
     8641 ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac cccaaggact
     8701 ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt
     8761 gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa
     8821 aatattctgt aacctttata agtaggcata acagttataa tcataacata ctgttttttc
     8881 ttactc
//
