LOCUS       sequence-258395-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP mRNA from the Drosophila Pepck1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  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 8508)
  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_pPepck1 eGFP SV40
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    147..268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1331..1347
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1332..1348
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1366..1368,1369..1371,1372..2085)
                     /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(1366..1368,1369..1371,1372..2082)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1411..1432)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1672..1691)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2019..2040
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2094..2215
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2925..2942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3096..3684)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3176..3195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3855..4646,4647..4715))
                     /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     4478..4497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4716..4820)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4888..4906
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6302..6581
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             6627..7652
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6627..7652
                     /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          8015..8080
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8210..8230
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ggttaataag gaatatttga tgtatagtgc cttgactaga gatcataatc agccatacca
       61 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
      121 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
      181 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
      241 gtttgtccaa actcatcaat gtatcttatc atgtctggat taattaaaaa aacagagcca
      301 tatttacgtg caactttccc gactgcttct gttgaacttt ttcttcgagt acccagagga
      361 ttctgggtct tctggtaaac tggattcggg gtacaaagtc acaccaccgg cagcaccttc
      421 atttctcctt ccccaaagac aaaaacaaga gtggccacat gacaaccatc cgggccatgc
      481 tattcttggt gcgcccccag agtcacatga atgtgaatac tggcctccaa ggcggccatg
      541 gccaattggg gaggcccccg gctcatctgg atgccgcgag gggggcggat tagcatacaa
      601 attttccgtc accgattcta caaataagcg aaagtttttt tcgatttggc aaatgtccaa
      661 aaacgagagg ccaacacatg tggtcaaaca ggcgtaagtg ggcgggcgcg tgtgatgtgg
      721 cccaccattc gatgttgcac caatgtagct atatttgtgg ccaatgcagc tatagacaca
      781 tccagaattt acccagctgc aggggacaac aactgctgtt gcctcagctg tccgtgaact
      841 ggtttctata ttcgcagcgg ccacatttgt atcgcataaa atcataaata gaataattaa
      901 attaacagcg cgaaagtggc aaatatcagg gtgttcaaaa taacaacaaa cgcttatcaa
      961 tgcaaacgca ctagaaaacg ctctcagcgc cacatgaaaa caacactctg ctgttgccat
     1021 gtggggacat accccatata cgtatctgta tgcccatgcc ccccattccg tagacccccc
     1081 cccccaccca caaccaactc atcgtagctg cgccgcctgt gaaaaaccaa aacaatcgcc
     1141 gagcctccgc cggcagccaa tcagctcgtc cggagtttga ccctctcact cggcgaggcg
     1201 atcgtaaaac ccgatttgcg tggcgatcgg ccgacttgtt ctagtcgcct cgatggccag
     1261 gcgtcgccgg ctgggtataa aaggccccgg atcggagcag ttggcatcag ttactcgtgg
     1321 ccagagtaac tcgaggtcga cggtatcgat aagcttgata tcaccatggt gagcaagggc
     1381 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1441 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1501 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1561 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1621 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1681 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1741 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1801 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1861 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1921 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1981 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2041 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagcggc cgcaacttgt
     2101 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2161 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaactag
     2221 tggatccact aggggccgcc gacgcgaggc tggatggcct tccccattat gattcttctc
     2281 gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac
     2341 gaccatcagg gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcatt
     2401 ggaccgctga tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca
     2461 tggattgtag gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg
     2521 agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc
     2581 caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc
     2641 agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg
     2701 ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct
     2761 ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg
     2821 actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg
     2881 tttcgtaaag tctggaaacg cggaagtcag cgctcttccg cttcctcgct cactgactcg
     2941 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3001 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3061 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3121 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3181 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3241 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3301 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3361 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3421 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3481 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3541 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3601 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3661 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3721 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3781 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3841 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3901 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3961 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4021 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4081 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4141 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     4201 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4261 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4321 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4381 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4441 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     4501 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4561 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4621 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4681 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4741 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4801 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4861 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
     4921 gaattcacat ttgtacgaat ttttttttta tcaaaagttc gagtttttca ccaatttcct
     4981 catcaaccga gcaaggcaaa cggctttgaa taatatggtg ttatatatac atatatcaaa
     5041 tcgctgctga ctgcgtgatt gatggcccca agattacata ttatcgaatc aggattcaga
     5101 aggagatcaa tgtcaaatgc ggacaggaac atgaaagacg cctgttatgc gcaattaaaa
     5161 atttgggttt aattgctgtg gaaactgttg ttggcggcat cttaagttcc tgtttaacaa
     5221 catcaactac ttatgtacgt agaagcgttt aagccatttg catacagatg agaactggct
     5281 tttgtgctaa tcagtcaaga tgactccgat gatgatgact cattacctga ccagttttcg
     5341 ctgctttctt ttcaacaact acttgtatat gtattgtatc caatagcaat acattgaatt
     5401 tccatggtct agtcacgtat tatcatttaa ttgacaccaa gtcgtgttat tgttgagcta
     5461 tcgagttcag ctcaaacatt tcttattccc atgaataagc cggcaaaaat atgcaatcta
     5521 tgaaagttaa tataagcaaa ccttactttg actcaatacc aatgcacttt gtgtcgatag
     5581 gttcacgcaa ttgaggcgat tattccgata acccaagcga ttgactgttc ccgtttcgat
     5641 tccaattgaa atttggaaat gtacaatagt tttgctatat gctgtcaagt acgctcttat
     5701 cttctctggg ttttcttcag agtttcgaaa cgcttcttct tttttttgtt tttttttttt
     5761 tggaatctcg tattttggaa ggggctcccc tctggaattt gttacactgt cgttatcatt
     5821 gcgaacaagc ggcccgaagc tatcagcgac tttaacattt acaatgcact tttttacgac
     5881 caattaaatg tacattttcc tttcttcgcc cgttgataag cgaacgcgat gtggcgcagg
     5941 caatgtgttg ctcttgcgac acaaacgcaa tcaaaatgga ttcaatttcg ctttttccca
     6001 gtgaaacgaa gaacgaaccg accatcatga tatgctcctc tgcatgttgc gtattgaatc
     6061 aatgacaatt tcaattaagc cgcccgttcg tcatgcgttt tcgtgcgctt cgaaatgctg
     6121 ataacgctgc tgtcctccaa ctgctttgca tgtggacaca attccattta tttaattctt
     6181 ttatttggat cggttaaatt aaaaagcgcc ttgttacgca tttaacgttg tttccggtgc
     6241 gtggtggttt catgcttctg ggaacggcaa atgggtttag gattgggaac ccctcatcat
     6301 ctgttggaat atactattca acctacaaaa gtaacgttaa acaacactac tttatatttg
     6361 atatgaatgg ccacaccttt tatgccataa aacatattgt aagagaatac cactcttttt
     6421 attccttctt tccttcttgt acgttttttg ctgtaagtag gtcgtggtgc tggtgttgca
     6481 gttgaaataa cttaaaatat aaatcataaa actcaaacat aaacttgact atttatttat
     6541 ttattaagaa aggaaatata aattataaat tacaacaggt tatggacctg cagccaagct
     6601 tggcgctcgt ccgggggcaa tgagatatga aaaagcctga actcaccgcg acgtctgtcg
     6661 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     6721 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     6781 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     6841 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     6901 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     6961 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     7021 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     7081 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     7141 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     7201 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     7261 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     7321 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     7381 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     7441 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     7501 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     7561 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     7621 gccccagcac tcgtccgagg gcaaaggaat agagtagatg ccgaccgaac aagagctgat
     7681 ttcgagaacg cctcagccag caactcgcgc gagcctagca aggcaaatgc gagagaacgg
     7741 ccttacgctt ggtggcacag ttctcgtcca cagttcgcta agctcgctcg gctgggtcgc
     7801 gggagggccg gtcgcagtga ttcaggccct tctggattgt gttggtcccc agggcacgat
     7861 tgtcatgccc acgcactcgg gtgatctgac tgatcccgca gattggagat cgccgcccgt
     7921 gcctgccgat tgggtgcaga tctttgtgaa ggaaccttac ttctgtggtg tgacataatt
     7981 ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt taagtgtata
     8041 atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct atggaactga
     8101 tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct cagaagaaat
     8161 gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc caaaaaagaa
     8221 gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt tgagtcatgc
     8281 tgtgtttagt aatagaactc ttgcttgctt tgctatttac accacaaagg aaaaagctgc
     8341 actgctatac aagaaaatta tggaaaaata ttctgtaacc tttataagta ggcataacag
     8401 ttataatcat aacatactgt tttttcttac tccacacagg catagagtgt ctgctattaa
     8461 taactatgct caaaaattgt gtacctttag ctttttaatt tgtaaagg
//
