LOCUS       sequence-254816-        7755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nanoLuciferase (nLuc) mRNA from the Drosophila 
            Metallothionein A promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7755)
  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 7755)
  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 nLuc SV40
FEATURES             Location/Qualifiers
     source          1..7755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(23..611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(103..122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(782..1573,1574..1642))
                     /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     1405..1424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1643..1747)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1815..1833
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3229..3508
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             3554..4579
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3554..4579
                     /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          4942..5007
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5137..5157
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5582..5716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        5717..6143
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     primer_bind     6151..6167
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6152..6168
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6186..6698
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             6699..6764
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    6776..6897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7607..7624)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
       61 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
      121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
      181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
      241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
      301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      421 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
      481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      781 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
      841 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      901 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
      961 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1021 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1081 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1141 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1201 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1261 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1321 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1381 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1441 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1501 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1561 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1621 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1681 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1741 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     1801 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttcacatttg
     1861 tacgaatttt ttttttatca aaagttcgag tttttcacca atttcctcat caaccgagca
     1921 aggcaaacgg ctttgaataa tatggtgtta tatatacata tatcaaatcg ctgctgactg
     1981 cgtgattgat ggccccaaga ttacatatta tcgaatcagg attcagaagg agatcaatgt
     2041 caaatgcgga caggaacatg aaagacgcct gttatgcgca attaaaaatt tgggtttaat
     2101 tgctgtggaa actgttgttg gcggcatctt aagttcctgt ttaacaacat caactactta
     2161 tgtacgtaga agcgtttaag ccatttgcat acagatgaga actggctttt gtgctaatca
     2221 gtcaagatga ctccgatgat gatgactcat tacctgacca gttttcgctg ctttcttttc
     2281 aacaactact tgtatatgta ttgtatccaa tagcaataca ttgaatttcc atggtctagt
     2341 cacgtattat catttaattg acaccaagtc gtgttattgt tgagctatcg agttcagctc
     2401 aaacatttct tattcccatg aataagccgg caaaaatatg caatctatga aagttaatat
     2461 aagcaaacct tactttgact caataccaat gcactttgtg tcgataggtt cacgcaattg
     2521 aggcgattat tccgataacc caagcgattg actgttcccg tttcgattcc aattgaaatt
     2581 tggaaatgta caatagtttt gctatatgct gtcaagtacg ctcttatctt ctctgggttt
     2641 tcttcagagt ttcgaaacgc ttcttctttt ttttgttttt ttttttttgg aatctcgtat
     2701 tttggaaggg gctcccctct ggaatttgtt acactgtcgt tatcattgcg aacaagcggc
     2761 ccgaagctat cagcgacttt aacatttaca atgcactttt ttacgaccaa ttaaatgtac
     2821 attttccttt cttcgcccgt tgataagcga acgcgatgtg gcgcaggcaa tgtgttgctc
     2881 ttgcgacaca aacgcaatca aaatggattc aatttcgctt tttcccagtg aaacgaagaa
     2941 cgaaccgacc atcatgatat gctcctctgc atgttgcgta ttgaatcaat gacaatttca
     3001 attaagccgc ccgttcgtca tgcgttttcg tgcgcttcga aatgctgata acgctgctgt
     3061 cctccaactg ctttgcatgt ggacacaatt ccatttattt aattctttta tttggatcgg
     3121 ttaaattaaa aagcgccttg ttacgcattt aacgttgttt ccggtgcgtg gtggtttcat
     3181 gcttctggga acggcaaatg ggtttaggat tgggaacccc tcatcatctg ttggaatata
     3241 ctattcaacc tacaaaagta acgttaaaca acactacttt atatttgata tgaatggcca
     3301 caccttttat gccataaaac atattgtaag agaataccac tctttttatt ccttctttcc
     3361 ttcttgtacg ttttttgctg taagtaggtc gtggtgctgg tgttgcagtt gaaataactt
     3421 aaaatataaa tcataaaact caaacataaa cttgactatt tatttattta ttaagaaagg
     3481 aaatataaat tataaattac aacaggttat ggacctgcag ccaagcttgg cgctcgtccg
     3541 ggggcaatga gatatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3601 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     3661 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     3721 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     3781 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     3841 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     3901 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     3961 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4021 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4081 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4141 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4201 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4261 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4321 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4381 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4441 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     4501 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4561 tccgagggca aaggaataga gtagatgccg accgaacaag agctgatttc gagaacgcct
     4621 cagccagcaa ctcgcgcgag cctagcaagg caaatgcgag agaacggcct tacgcttggt
     4681 ggcacagttc tcgtccacag ttcgctaagc tcgctcggct gggtcgcggg agggccggtc
     4741 gcagtgattc aggcccttct ggattgtgtt ggtccccagg gcacgattgt catgcccacg
     4801 cactcgggtg atctgactga tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg
     4861 gtgcagatct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
     4921 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
     4981 ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag
     5041 tggtggaatg cctttaatga ggaaaacctg ttttgctcag aagaaatgcc atctagtgat
     5101 gatgaggcta ctgctgactc tcaacattct actcctccaa aaaagaagag aaaggtagaa
     5161 gaccccaagg actttccttc agaattgcta agttttttga gtcatgctgt gtttagtaat
     5221 agaactcttg cttgctttgc tatttacacc acaaaggaaa aagctgcact gctatacaag
     5281 aaaattatgg aaaaatattc tgtaaccttt ataagtaggc ataacagtta taatcataac
     5341 atactgtttt ttcttactcc acacaggcat agagtgtctg ctattaataa ctatgctcaa
     5401 aaattgtgta cctttagctt tttaatttgt aaaggggtta ataaggaata tttgatgtat
     5461 agtgccttga ctagagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     5521 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     5581 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5701 ttatcatgtc tggatcaatt cgttgcagga caggatgtgg tgcccgatgt gactagctct
     5761 ttgctgcagg ccgtcctatc ctctggttcc gataagagac ccagaactcc ggccccccac
     5821 cgcccaccgc cacccccata catatgtggt acgcaagtaa gagtgcctgc gcatgcccca
     5881 tgtgccccac caagagcttt gcatcccata caagtcccca aagtggagaa ccgaaccaat
     5941 tcttcgcggg cagaacaaaa gcttctgcac acgtctccac tcgaatttgg agccggccgg
     6001 cgtgtgcaaa agaggtgaat cgaacgaaag acccgtgtgt aaagccgcgt ttccaaaatg
     6061 tataaaaccg agagcatctg gccaatgtgc atcagttgtg gtcagcagca aaatcaagtg
     6121 aatcatctca gtgcaactaa aggggggatc tcgaggtcga cggtatcgat aagcttgata
     6181 tcaccatggt cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca
     6241 acctggacca agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt
     6301 ccgtaactcc gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc
     6361 atgtcatcat cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt
     6421 ttaaggtggt gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac
     6481 tggtaatcga cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca
     6541 tcgccgtgtt cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa
     6601 ttatcgacga gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg
     6661 gagtgaccgg ctggcggctg tgcgaacgca ttctggcgga ctacaaagac catgacggtg
     6721 attataaaga tcatgacatc gattacaagg atgacgatga caagtaagcg gccgcaactt
     6781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6841 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaact
     6901 agtggatcca ctaggggccg ccgacgcgag gctggatggc cttccccatt atgattcttc
     6961 tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg
     7021 acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta acttcgatca
     7081 ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg aacgggttgg
     7141 catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat
     7201 ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccac
     7261 tccaagaatt ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg
     7321 gcagaacata tccatcgcgt ccgccatctc cagcagccgc acgcggcgca tctcgggcag
     7381 cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     7441 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     7501 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     7561 tgtttcgtaa agtctggaaa cgcggaagtc agcgctcttc cgcttcctcg ctcactgact
     7621 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     7681 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     7741 aggccaggaa ccgta
//
