LOCUS       sequence-257549-        7933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an eGFP mRNA from the Drosophila Metallothionein A 
            promoter that ends in the MALAT1 triple helix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7933)
  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 7933)
  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 eGFP MALAT1
FEATURES             Location/Qualifiers
     source          1..7933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    283..417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(320..339)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     374..393
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        418..844
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     primer_bind     852..868
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     853..869
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(887..889,890..892,893..1606)
                     /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(887..889,890..892,893..1603)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(932..953)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1193..1212)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1540..1561
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_RNA        1725..1776
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     primer_bind     complement(2486..2503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2657..3245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2737..2756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3416..4207,4208..4276))
                     /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     4039..4058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4277..4381)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4449..4467
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5863..6142
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             6188..7213
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6188..7213
                     /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          7576..7641
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7771..7791
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctgtaacctt tataagtagg cataacagtt ataatcataa catactgttt tttcttactc
       61 cacacaggca tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct
      121 ttttaatttg taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc
      181 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
      241 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
      301 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
      361 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaat
      421 tcgttgcagg acaggatgtg gtgcccgatg tgactagctc tttgctgcag gccgtcctat
      481 cctctggttc cgataagaga cccagaactc cggcccccca ccgcccaccg ccacccccat
      541 acatatgtgg tacgcaagta agagtgcctg cgcatgcccc atgtgcccca ccaagagctt
      601 tgcatcccat acaagtcccc aaagtggaga accgaaccaa ttcttcgcgg gcagaacaaa
      661 agcttctgca cacgtctcca ctcgaatttg gagccggccg gcgtgtgcaa aagaggtgaa
      721 tcgaacgaaa gacccgtgtg taaagccgcg tttccaaaat gtataaaacc gagagcatct
      781 ggccaatgtg catcagttgt ggtcagcagc aaaatcaagt gaatcatctc agtgcaacta
      841 aaggggggat ctcgaggtcg acggtatcga taagcttgat atcaccatgg tgagcaaggg
      901 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      961 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1021 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1081 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1141 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1201 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1261 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1321 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1381 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1441 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1501 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1561 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagcgg ccgcgattcg
     1621 tcagtagggt tgtaaaggtt tttcttttcc tgagaaaaca accttttgtt ttctcaggtt
     1681 ttgctttttg gcctttccct agctttaaaa aaaaaaaagc aaaacctgtc accggatgtg
     1741 ttttccggtc tgatgagtcc gtgaggacga aacaggacta gtggatccac taggggccgc
     1801 cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga
     1861 tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc
     1921 aaggatcgct cgcggctctt accagcctaa cttcgatcat tggaccgctg atcgtcacgg
     1981 cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc
     2041 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
     2101 gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca
     2161 attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc
     2221 cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt
     2281 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     2341 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     2401 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
     2461 gcggaagtca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2521 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2581 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2641 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2701 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2761 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2821 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2881 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2941 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3001 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3061 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     3121 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3181 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3241 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3301 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3361 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3421 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3481 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3541 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3601 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3661 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3721 ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3781 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3841 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3901 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3961 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4021 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4081 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4141 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4201 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4261 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4321 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4381 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     4441 cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattcaca tttgtacgaa
     4501 tttttttttt atcaaaagtt cgagtttttc accaatttcc tcatcaaccg agcaaggcaa
     4561 acggctttga ataatatggt gttatatata catatatcaa atcgctgctg actgcgtgat
     4621 tgatggcccc aagattacat attatcgaat caggattcag aaggagatca atgtcaaatg
     4681 cggacaggaa catgaaagac gcctgttatg cgcaattaaa aatttgggtt taattgctgt
     4741 ggaaactgtt gttggcggca tcttaagttc ctgtttaaca acatcaacta cttatgtacg
     4801 tagaagcgtt taagccattt gcatacagat gagaactggc ttttgtgcta atcagtcaag
     4861 atgactccga tgatgatgac tcattacctg accagttttc gctgctttct tttcaacaac
     4921 tacttgtata tgtattgtat ccaatagcaa tacattgaat ttccatggtc tagtcacgta
     4981 ttatcattta attgacacca agtcgtgtta ttgttgagct atcgagttca gctcaaacat
     5041 ttcttattcc catgaataag ccggcaaaaa tatgcaatct atgaaagtta atataagcaa
     5101 accttacttt gactcaatac caatgcactt tgtgtcgata ggttcacgca attgaggcga
     5161 ttattccgat aacccaagcg attgactgtt cccgtttcga ttccaattga aatttggaaa
     5221 tgtacaatag ttttgctata tgctgtcaag tacgctctta tcttctctgg gttttcttca
     5281 gagtttcgaa acgcttcttc ttttttttgt tttttttttt ttggaatctc gtattttgga
     5341 aggggctccc ctctggaatt tgttacactg tcgttatcat tgcgaacaag cggcccgaag
     5401 ctatcagcga ctttaacatt tacaatgcac ttttttacga ccaattaaat gtacattttc
     5461 ctttcttcgc ccgttgataa gcgaacgcga tgtggcgcag gcaatgtgtt gctcttgcga
     5521 cacaaacgca atcaaaatgg attcaatttc gctttttccc agtgaaacga agaacgaacc
     5581 gaccatcatg atatgctcct ctgcatgttg cgtattgaat caatgacaat ttcaattaag
     5641 ccgcccgttc gtcatgcgtt ttcgtgcgct tcgaaatgct gataacgctg ctgtcctcca
     5701 actgctttgc atgtggacac aattccattt atttaattct tttatttgga tcggttaaat
     5761 taaaaagcgc cttgttacgc atttaacgtt gtttccggtg cgtggtggtt tcatgcttct
     5821 gggaacggca aatgggttta ggattgggaa cccctcatca tctgttggaa tatactattc
     5881 aacctacaaa agtaacgtta aacaacacta ctttatattt gatatgaatg gccacacctt
     5941 ttatgccata aaacatattg taagagaata ccactctttt tattccttct ttccttcttg
     6001 tacgtttttt gctgtaagta ggtcgtggtg ctggtgttgc agttgaaata acttaaaata
     6061 taaatcataa aactcaaaca taaacttgac tatttattta tttattaaga aaggaaatat
     6121 aaattataaa ttacaacagg ttatggacct gcagccaagc ttggcgctcg tccgggggca
     6181 atgagatatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     6241 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     6301 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     6361 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     6421 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     6481 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     6541 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     6601 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     6661 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     6721 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     6781 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     6841 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     6901 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     6961 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     7021 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     7081 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     7141 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     7201 ggcaaaggaa tagagtagat gccgaccgaa caagagctga tttcgagaac gcctcagcca
     7261 gcaactcgcg cgagcctagc aaggcaaatg cgagagaacg gccttacgct tggtggcaca
     7321 gttctcgtcc acagttcgct aagctcgctc ggctgggtcg cgggagggcc ggtcgcagtg
     7381 attcaggccc ttctggattg tgttggtccc cagggcacga ttgtcatgcc cacgcactcg
     7441 ggtgatctga ctgatcccgc agattggaga tcgccgcccg tgcctgccga ttgggtgcag
     7501 atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag
     7561 atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt
     7621 ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg
     7681 aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag
     7741 gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc
     7801 aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact
     7861 cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt
     7921 atggaaaaat att
//
