LOCUS       sequence-258479-        7851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an eGFP mRNA from the Drosophila Metallothionein A 
            promoter that has 20 nt of the MtnA 5' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7851)
  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 7851)
  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_pMtnA UTR delta 21-55 eGFP SV40
FEATURES             Location/Qualifiers
     source          1..7851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(140..159)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     487..508
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    562..683
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1393..1410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1564..2152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1644..1663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2323..3114,3115..3183))
                     /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     2946..2965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3184..3288)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3356..3374
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4770..5049
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             5095..6120
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5095..6120
                     /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          6483..6548
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             6678..6698
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7123..7244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     7650..7666
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7651..7667
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(7685..7687,7688..7690,7691..7851,1..553)
                     /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(7685..7687,7688..7690,7691..7851,1..550)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7730..7751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
       61 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      121 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      181 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      241 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
      301 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      361 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      421 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
      481 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
      541 gctgtacaag taagcggccg caacttgttt attgcagctt ataatggtta caaataaagc
      601 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
      661 tccaaactca tcaatgtatc ttaactagtg gatccactag gggccgccga cgcgaggctg
      721 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
      781 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc
      841 ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
      901 ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg
      961 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
     1021 cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga
     1081 actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc
     1141 agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg
     1201 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa
     1261 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca
     1321 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg
     1381 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1801 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1981 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
     2641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     2941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3301 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3361 gcgtatcacg aggccctttc gtcttcaaga attcacattt gtacgaattt tttttttatc
     3421 aaaagttcga gtttttcacc aatttcctca tcaaccgagc aaggcaaacg gctttgaata
     3481 atatggtgtt atatatacat atatcaaatc gctgctgact gcgtgattga tggccccaag
     3541 attacatatt atcgaatcag gattcagaag gagatcaatg tcaaatgcgg acaggaacat
     3601 gaaagacgcc tgttatgcgc aattaaaaat ttgggtttaa ttgctgtgga aactgttgtt
     3661 ggcggcatct taagttcctg tttaacaaca tcaactactt atgtacgtag aagcgtttaa
     3721 gccatttgca tacagatgag aactggcttt tgtgctaatc agtcaagatg actccgatga
     3781 tgatgactca ttacctgacc agttttcgct gctttctttt caacaactac ttgtatatgt
     3841 attgtatcca atagcaatac attgaatttc catggtctag tcacgtatta tcatttaatt
     3901 gacaccaagt cgtgttattg ttgagctatc gagttcagct caaacatttc ttattcccat
     3961 gaataagccg gcaaaaatat gcaatctatg aaagttaata taagcaaacc ttactttgac
     4021 tcaataccaa tgcactttgt gtcgataggt tcacgcaatt gaggcgatta ttccgataac
     4081 ccaagcgatt gactgttccc gtttcgattc caattgaaat ttggaaatgt acaatagttt
     4141 tgctatatgc tgtcaagtac gctcttatct tctctgggtt ttcttcagag tttcgaaacg
     4201 cttcttcttt tttttgtttt tttttttttg gaatctcgta ttttggaagg ggctcccctc
     4261 tggaatttgt tacactgtcg ttatcattgc gaacaagcgg cccgaagcta tcagcgactt
     4321 taacatttac aatgcacttt tttacgacca attaaatgta cattttcctt tcttcgcccg
     4381 ttgataagcg aacgcgatgt ggcgcaggca atgtgttgct cttgcgacac aaacgcaatc
     4441 aaaatggatt caatttcgct ttttcccagt gaaacgaaga acgaaccgac catcatgata
     4501 tgctcctctg catgttgcgt attgaatcaa tgacaatttc aattaagccg cccgttcgtc
     4561 atgcgttttc gtgcgcttcg aaatgctgat aacgctgctg tcctccaact gctttgcatg
     4621 tggacacaat tccatttatt taattctttt atttggatcg gttaaattaa aaagcgcctt
     4681 gttacgcatt taacgttgtt tccggtgcgt ggtggtttca tgcttctggg aacggcaaat
     4741 gggtttagga ttgggaaccc ctcatcatct gttggaatat actattcaac ctacaaaagt
     4801 aacgttaaac aacactactt tatatttgat atgaatggcc acacctttta tgccataaaa
     4861 catattgtaa gagaatacca ctctttttat tccttctttc cttcttgtac gttttttgct
     4921 gtaagtaggt cgtggtgctg gtgttgcagt tgaaataact taaaatataa atcataaaac
     4981 tcaaacataa acttgactat ttatttattt attaagaaag gaaatataaa ttataaatta
     5041 caacaggtta tggacctgca gccaagcttg gcgctcgtcc gggggcaatg agatatgaaa
     5101 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     5161 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     5221 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     5281 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
     5341 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     5401 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
     5461 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     5521 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     5581 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     5641 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
     5701 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     5761 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
     5821 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
     5881 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     5941 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     6001 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
     6061 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag
     6121 agtagatgcc gaccgaacaa gagctgattt cgagaacgcc tcagccagca actcgcgcga
     6181 gcctagcaag gcaaatgcga gagaacggcc ttacgcttgg tggcacagtt ctcgtccaca
     6241 gttcgctaag ctcgctcggc tgggtcgcgg gagggccggt cgcagtgatt caggcccttc
     6301 tggattgtgt tggtccccag ggcacgattg tcatgcccac gcactcgggt gatctgactg
     6361 atcccgcaga ttggagatcg ccgcccgtgc ctgccgattg ggtgcagatc tttgtgaagg
     6421 aaccttactt ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta
     6481 aggtaaatat aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg
     6541 tattttagat tccaacctat ggaactgatg aatgggagca gtggtggaat gcctttaatg
     6601 aggaaaacct gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact
     6661 ctcaacattc tactcctcca aaaaagaaga gaaaggtaga agaccccaag gactttcctt
     6721 cagaattgct aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg
     6781 ctatttacac cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt
     6841 ctgtaacctt tataagtagg cataacagtt ataatcataa catactgttt tttcttactc
     6901 cacacaggca tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct
     6961 ttttaatttg taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc
     7021 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     7081 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     7141 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     7201 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggattaat
     7261 taattgcagg acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat
     7321 cctctggttc cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat
     7381 acatatgtgg tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt
     7441 tgcatcccat acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa
     7501 agcttctgca cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa
     7561 tcgaacgaaa gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct
     7621 ggccaatgtg catcagttgt ggtcagcact cgaggtcgac ggtatcgata agcttgatat
     7681 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     7741 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     7801 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg t
//
