LOCUS       sequence-258445-        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an eGFP mRNA from the Drosophila Metallothionein A 
            promoter that has 40 nt of the MtnA 5' UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7871)
  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 7871)
  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 41-55 eGFP SV40
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..157
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     397..418
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(481..497)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(482..498)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    924..1045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1470..1490)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1620..1685
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(2048..3073)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(2048..3073)
                     /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(3119..3398)
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     primer_bind     complement(4794..4812)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4880..4984
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4985..5053,5054..5845)
                     /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(5203..5222)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6016..6604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6505..6524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6758..6775
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7485..7606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(7615..7871,1..457,458..460,461..463))
                     /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(7618..7871,1..457,458..460,461..463))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7660..7681)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
       61 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
      121 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
      181 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
      241 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
      301 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
      361 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
      421 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catggtgata tcaagcttat
      481 cgataccgtc gacctcgagg atgattcact tgattttgct gctgaccaca actgatgcac
      541 attggccaga tgctctcggt tttatacatt ttggaaacgc ggctttacac acgggtcttt
      601 cgttcgattc acctcttttg cacacgccgg ccggctccaa attcgagtgg agacgtgtgc
      661 agaagctttt gttctgcccg cgaagaattg gttcggttct ccactttggg gacttgtatg
      721 ggatgcaaag ctcttggtgg ggcacatggg gcatgcgcag gcactcttac ttgcgtacca
      781 catatgtatg ggggtggcgg tgggcggtgg ggggccggag ttctgggtct cttatcggaa
      841 ccagaggata ggacggcctg cagcaaagag ctagtcacat cgggcaccac atcctgtcct
      901 gcaattaatt aatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
      961 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1021 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1081 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     1141 tgattatgat ctctagtcaa ggcactatac atcaaatatt ccttattaac ccctttacaa
     1201 attaaaaagc taaaggtaca caatttttga gcatagttat taatagcaga cactctatgc
     1261 ctgtgtggag taagaaaaaa cagtatgtta tgattataac tgttatgcct acttataaag
     1321 gttacagaat atttttccat aattttcttg tatagcagtg cagctttttc ctttgtggtg
     1381 taaatagcaa agcaagcaag agttctatta ctaaacacag catgactcaa aaaacttagc
     1441 aattctgaag gaaagtcctt ggggtcttct acctttctct tcttttttgg aggagtagaa
     1501 tgttgagagt cagcagtagc ctcatcatca ctagatggca tttcttctga gcaaaacagg
     1561 ttttcctcat taaaggcatt ccaccactgc tcccattcat cagttccata ggttggaatc
     1621 taaaatacac aaacaattag aatcagtagt ttaacacatt atacacttaa aaattttata
     1681 tttaccttag agctttaaat ctctgtaggt agtttgtcca attatgtcac accacagaag
     1741 taaggttcct tcacaaagat ctgcacccaa tcggcaggca cgggcggcga tctccaatct
     1801 gcgggatcag tcagatcacc cgagtgcgtg ggcatgacaa tcgtgccctg gggaccaaca
     1861 caatccagaa gggcctgaat cactgcgacc ggccctcccg cgacccagcc gagcgagctt
     1921 agcgaactgt ggacgagaac tgtgccacca agcgtaaggc cgttctctcg catttgcctt
     1981 gctaggctcg cgcgagttgc tggctgaggc gttctcgaaa tcagctcttg ttcggtcggc
     2041 atctactcta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca ctatcggcga
     2101 gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc
     2161 cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc caagctgcat
     2221 catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg cggagcatat
     2281 acgcccggag ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct
     2341 gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc tcgtattggg
     2401 aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca
     2461 ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg
     2521 cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg tcgtccatca
     2581 cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca cgccatgtag
     2641 tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg ctaagatcgg
     2701 ccgcagcgat cgcatccatg gcctccgcga ccggctgcag aacagcgggc agttcggttt
     2761 caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag gtcaggctct
     2821 cgctgaattc cccaatgtca agcacttccg gaatcgggag cgcggccgat gcaaagtgcc
     2881 gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc aggacatatc
     2941 cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag agctgcatca
     3001 ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc gcggtgagtt
     3061 caggcttttt catatctcat tgcccccgga cgagcgccaa gcttggctgc aggtccataa
     3121 cctgttgtaa tttataattt atatttcctt tcttaataaa taaataaata gtcaagttta
     3181 tgtttgagtt ttatgattta tattttaagt tatttcaact gcaacaccag caccacgacc
     3241 tacttacagc aaaaaacgta caagaaggaa agaaggaata aaaagagtgg tattctctta
     3301 caatatgttt tatggcataa aaggtgtggc cattcatatc aaatataaag tagtgttgtt
     3361 taacgttact tttgtaggtt gaatagtata ttccaacaga tgatgagggg ttcccaatcc
     3421 taaacccatt tgccgttccc agaagcatga aaccaccacg caccggaaac aacgttaaat
     3481 gcgtaacaag gcgcttttta atttaaccga tccaaataaa agaattaaat aaatggaatt
     3541 gtgtccacat gcaaagcagt tggaggacag cagcgttatc agcatttcga agcgcacgaa
     3601 aacgcatgac gaacgggcgg cttaattgaa attgtcattg attcaatacg caacatgcag
     3661 aggagcatat catgatggtc ggttcgttct tcgtttcact gggaaaaagc gaaattgaat
     3721 ccattttgat tgcgtttgtg tcgcaagagc aacacattgc ctgcgccaca tcgcgttcgc
     3781 ttatcaacgg gcgaagaaag gaaaatgtac atttaattgg tcgtaaaaaa gtgcattgta
     3841 aatgttaaag tcgctgatag cttcgggccg cttgttcgca atgataacga cagtgtaaca
     3901 aattccagag gggagcccct tccaaaatac gagattccaa aaaaaaaaaa acaaaaaaaa
     3961 gaagaagcgt ttcgaaactc tgaagaaaac ccagagaaga taagagcgta cttgacagca
     4021 tatagcaaaa ctattgtaca tttccaaatt tcaattggaa tcgaaacggg aacagtcaat
     4081 cgcttgggtt atcggaataa tcgcctcaat tgcgtgaacc tatcgacaca aagtgcattg
     4141 gtattgagtc aaagtaaggt ttgcttatat taactttcat agattgcata tttttgccgg
     4201 cttattcatg ggaataagaa atgtttgagc tgaactcgat agctcaacaa taacacgact
     4261 tggtgtcaat taaatgataa tacgtgacta gaccatggaa attcaatgta ttgctattgg
     4321 atacaataca tatacaagta gttgttgaaa agaaagcagc gaaaactggt caggtaatga
     4381 gtcatcatca tcggagtcat cttgactgat tagcacaaaa gccagttctc atctgtatgc
     4441 aaatggctta aacgcttcta cgtacataag tagttgatgt tgttaaacag gaacttaaga
     4501 tgccgccaac aacagtttcc acagcaatta aacccaaatt tttaattgcg cataacaggc
     4561 gtctttcatg ttcctgtccg catttgacat tgatctcctt ctgaatcctg attcgataat
     4621 atgtaatctt ggggccatca atcacgcagt cagcagcgat ttgatatatg tatatataac
     4681 accatattat tcaaagccgt ttgccttgct cggttgatga ggaaattggt gaaaaactcg
     4741 aacttttgat aaaaaaaaaa ttcgtacaaa tgtgaattct tgaagacgaa agggcctcgt
     4801 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4861 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4921 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4981 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5041 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5101 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5161 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5221 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5281 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5341 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5401 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5461 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5521 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5581 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5641 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5701 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5761 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5821 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5881 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5941 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6001 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6061 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6121 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6181 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6241 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6301 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6361 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6421 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6481 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6541 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6601 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6661 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6721 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6781 ggaagagcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     6841 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     6901 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     6961 acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgag atgcgccgcg
     7021 tgcggctgct ggagatggcg gacgcgatgg atatgttctg ccaagggttg gtttgcgcat
     7081 tcacagttct ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga
     7141 ggtgccgccg gcttccattc aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc
     7201 ggggaggcag acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct
     7261 cgccgaggcg gcataaatcg ccgtgacgat cagcggtcca atgatcgaag ttaggctggt
     7321 aagagccgcg agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag
     7381 catggcctgc aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa
     7441 ggccatccag cctcgcgtcg gcggccccta gtggatccac tagttaagat acattgatga
     7501 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     7561 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgcgg ccgcttactt
     7621 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     7681 gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt
     7741 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
     7801 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
     7861 ctgcttgtcg g
//
