LOCUS       sequence-258399-        7886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an eGFP mRNA from the Drosophila Metallothionein A 
            promoter that has 55 nt of the MtnA 5' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7886)
  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 7886)
  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 1-55 eGFP SV40
FEATURES             Location/Qualifiers
     source          1..7886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     186..205
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     445..466
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(529..545)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(530..546)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(548..973)
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     polyA_signal    987..1108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1533..1553)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1683..1748
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(2111..3136)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(2111..3136)
                     /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"
     promoter        complement(3182..3461)
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     primer_bind     complement(4857..4875)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4943..5047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5048..5116,5117..5908)
                     /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     complement(5266..5285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6079..6667
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6568..6587
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6821..6838
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7548..7669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(7678..7886,1..505,506..508,509..511))
                     /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             complement(join(7681..7886,1..505,506..508,509..511))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7723..7744)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
       61 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
      121 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
      181 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
      241 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
      301 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
      361 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
      421 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
      481 cggtgaacag ctcctcgccc ttgctcacca tggtgatatc aagcttatcg ataccgtcga
      541 cctcgagctt tagttgcact gagatgattc acttgatttt gctgctgacc acaactgatg
      601 cacattggcc agatgctctc ggttttatac attttggaaa cgcggcttta cacacgggtc
      661 tttcgttcga ttcacctctt ttgcacacgc cggccggctc caaattcgag tggagacgtg
      721 tgcagaagct tttgttctgc ccgcgaagaa ttggttcggt tctccacttt ggggacttgt
      781 atgggatgca aagctcttgg tggggcacat ggggcatgcg caggcactct tacttgcgta
      841 ccacatatgt atgggggtgg cggtgggcgg tggggggccg gagttctggg tctcttatcg
      901 gaaccagagg ataggacggc ctgcagcaaa gagctagtca catcgggcac cacatcctgt
      961 cctgcaatta attaatccag acatgataag atacattgat gagtttggac aaaccacaac
     1021 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     1081 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     1141 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     1201 ggctgattat gatctctagt caaggcacta tacatcaaat attccttatt aaccccttta
     1261 caaattaaaa agctaaaggt acacaatttt tgagcatagt tattaatagc agacactcta
     1321 tgcctgtgtg gagtaagaaa aaacagtatg ttatgattat aactgttatg cctacttata
     1381 aaggttacag aatatttttc cataattttc ttgtatagca gtgcagcttt ttcctttgtg
     1441 gtgtaaatag caaagcaagc aagagttcta ttactaaaca cagcatgact caaaaaactt
     1501 agcaattctg aaggaaagtc cttggggtct tctacctttc tcttcttttt tggaggagta
     1561 gaatgttgag agtcagcagt agcctcatca tcactagatg gcatttcttc tgagcaaaac
     1621 aggttttcct cattaaaggc attccaccac tgctcccatt catcagttcc ataggttgga
     1681 atctaaaata cacaaacaat tagaatcagt agtttaacac attatacact taaaaatttt
     1741 atatttacct tagagcttta aatctctgta ggtagtttgt ccaattatgt cacaccacag
     1801 aagtaaggtt ccttcacaaa gatctgcacc caatcggcag gcacgggcgg cgatctccaa
     1861 tctgcgggat cagtcagatc acccgagtgc gtgggcatga caatcgtgcc ctggggacca
     1921 acacaatcca gaagggcctg aatcactgcg accggccctc ccgcgaccca gccgagcgag
     1981 cttagcgaac tgtggacgag aactgtgcca ccaagcgtaa ggccgttctc tcgcatttgc
     2041 cttgctaggc tcgcgcgagt tgctggctga ggcgttctcg aaatcagctc ttgttcggtc
     2101 ggcatctact ctattccttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg
     2161 cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac
     2221 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg
     2281 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca
     2341 tatacgcccg gagccgcggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg
     2401 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt
     2461 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg
     2521 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct
     2581 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca
     2641 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg
     2701 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat
     2761 cggccgcagc gatcgcatcc atggcctccg cgaccggctg cagaacagcg ggcagttcgg
     2821 tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg ggagatgcaa taggtcaggc
     2881 tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt
     2941 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat
     3001 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca
     3061 tcaggtcgga gacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga
     3121 gttcaggctt tttcatatct cattgccccc ggacgagcgc caagcttggc tgcaggtcca
     3181 taacctgttg taatttataa tttatatttc ctttcttaat aaataaataa atagtcaagt
     3241 ttatgtttga gttttatgat ttatatttta agttatttca actgcaacac cagcaccacg
     3301 acctacttac agcaaaaaac gtacaagaag gaaagaagga ataaaaagag tggtattctc
     3361 ttacaatatg ttttatggca taaaaggtgt ggccattcat atcaaatata aagtagtgtt
     3421 gtttaacgtt acttttgtag gttgaatagt atattccaac agatgatgag gggttcccaa
     3481 tcctaaaccc atttgccgtt cccagaagca tgaaaccacc acgcaccgga aacaacgtta
     3541 aatgcgtaac aaggcgcttt ttaatttaac cgatccaaat aaaagaatta aataaatgga
     3601 attgtgtcca catgcaaagc agttggagga cagcagcgtt atcagcattt cgaagcgcac
     3661 gaaaacgcat gacgaacggg cggcttaatt gaaattgtca ttgattcaat acgcaacatg
     3721 cagaggagca tatcatgatg gtcggttcgt tcttcgtttc actgggaaaa agcgaaattg
     3781 aatccatttt gattgcgttt gtgtcgcaag agcaacacat tgcctgcgcc acatcgcgtt
     3841 cgcttatcaa cgggcgaaga aaggaaaatg tacatttaat tggtcgtaaa aaagtgcatt
     3901 gtaaatgtta aagtcgctga tagcttcggg ccgcttgttc gcaatgataa cgacagtgta
     3961 acaaattcca gaggggagcc ccttccaaaa tacgagattc caaaaaaaaa aaaacaaaaa
     4021 aaagaagaag cgtttcgaaa ctctgaagaa aacccagaga agataagagc gtacttgaca
     4081 gcatatagca aaactattgt acatttccaa atttcaattg gaatcgaaac gggaacagtc
     4141 aatcgcttgg gttatcggaa taatcgcctc aattgcgtga acctatcgac acaaagtgca
     4201 ttggtattga gtcaaagtaa ggtttgctta tattaacttt catagattgc atatttttgc
     4261 cggcttattc atgggaataa gaaatgtttg agctgaactc gatagctcaa caataacacg
     4321 acttggtgtc aattaaatga taatacgtga ctagaccatg gaaattcaat gtattgctat
     4381 tggatacaat acatatacaa gtagttgttg aaaagaaagc agcgaaaact ggtcaggtaa
     4441 tgagtcatca tcatcggagt catcttgact gattagcaca aaagccagtt ctcatctgta
     4501 tgcaaatggc ttaaacgctt ctacgtacat aagtagttga tgttgttaaa caggaactta
     4561 agatgccgcc aacaacagtt tccacagcaa ttaaacccaa atttttaatt gcgcataaca
     4621 ggcgtctttc atgttcctgt ccgcatttga cattgatctc cttctgaatc ctgattcgat
     4681 aatatgtaat cttggggcca tcaatcacgc agtcagcagc gatttgatat atgtatatat
     4741 aacaccatat tattcaaagc cgtttgcctt gctcggttga tgaggaaatt ggtgaaaaac
     4801 tcgaactttt gataaaaaaa aaattcgtac aaatgtgaat tcttgaagac gaaagggcct
     4861 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4921 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4981 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5041 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5101 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5161 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5221 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5281 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5341 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5401 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5461 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5521 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5581 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5641 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5701 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5761 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5821 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5881 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5941 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6001 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6061 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6121 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6181 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     6241 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6301 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6361 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6421 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6481 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6541 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6601 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6661 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6721 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6781 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6841 agcggaagag cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     6901 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     6961 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     7021 acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc
     7081 gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg
     7141 cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag
     7201 cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa
     7261 cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt
     7321 gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct
     7381 ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga
     7441 cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg
     7501 gaaggccatc cagcctcgcg tcggcggccc ctagtggatc cactagttaa gatacattga
     7561 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     7621 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg cggccgctta
     7681 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
     7741 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg
     7801 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     7861 gagctgcacg ctgccgtcct cgatgt
//
