LOCUS       sequence-416778-        7764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Drosophila IntS7 from inducible MtnA promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7764)
  AUTHORS   Fujiwara R, Zhai SN, Liang D, Shah AP, Tracey M, Ma XK, Fields CJ, 
            Mendoza-Figueroa MS, Meline MC, Tatomer DC, Yang L, Wilusz JE
  TITLE     IntS6 and the Integrator phosphatase module tune the efficiency of 
            select premature transcription termination events.
  JOURNAL   Mol Cell. 2023 Nov 14:S1097-2765(23)00902-4. doi: 
            10.1016/j.molcel.2023.10.035.
  PUBMED    37995689
REFERENCE   2  (bases 1 to 7764)
  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: "Mol Cell. 
            2023 Nov 14:S1097-2765(23)00902-4. doi: 
            10.1016/j.molcel.2023.10.035."
COMMENT     Sequence Label: pMtnA FLAG-IntS7 puro
FEATURES             Location/Qualifiers
     source          1..7764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..115
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(729..745)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(729..745)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(742..764)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    753..769
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(777..783,784..801,802..807))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    822..843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(960..977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1131..1719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1211..1230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1890..2681,2682..2750))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2513..2532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2751..2855)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2979..3001)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3101..3120
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3314..3336
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3328..3345
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3329..3345
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3351..3777
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3802..3867
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6914..6955
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6965..6982
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7005..7022)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7024..7158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7048..7067)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7102..7121
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(7280..7764,1..115))
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7363..7383)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggtcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg
       61 cccgggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggaag
      121 gtcgtctcct tgtgaggggt caggggcgtg ggtcagggga tggtggcggc accggtcgtg
      181 gcggccgacc tgcaggcatg caagctatcc aattcctgca gcccggggga tctgttgtaa
      241 tttataattt atatttcctt tcttaataaa taaataaata gtcaagttta tgtttgagtt
      301 ttatgattta tattttaagt tatttcaact gcaacaccag caccacgacc tacttacagc
      361 aaaaaacgta caagaaggaa agaaggaata aaaagagtgg tattctctta caatatgttt
      421 tatggcataa aaggtgtggc cattcatatc aaatataaag tagtgttgtt taacgttact
      481 tttgtaggtt gaatagtata ttccaacaga tgatgagggg ttcccaatcc taaacccatt
      541 tgccgttccc agaagcatga aaccaccacg cacccgatcc tctaggacaa caacaattgc
      601 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      661 ctctacaaat gtggtatggc tgattatgat cagtcgacct gcaggcatgc aagcttggcg
      721 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
      781 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
      841 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
      901 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
      961 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1021 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1081 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1141 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1201 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1261 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1321 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1381 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1441 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1501 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1561 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1621 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1681 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1741 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1801 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1861 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1921 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     1981 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2041 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2101 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2161 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2221 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2281 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2341 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2401 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2461 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2521 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2581 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2641 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2701 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2761 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2821 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2881 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2941 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3001 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3061 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     3121 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     3181 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     3241 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     3301 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt aattcgttgc
     3361 aggacaggat gtggtgcccg atgtgactag ctctttgctg caggccgtcc tatcctctgg
     3421 ttccgataag agacccagaa ctccggcccc ccaccgccca ccgccacccc catacatatg
     3481 tggtacgcaa gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag ctttgcatcc
     3541 catacaagtc cccaaagtgg agaaccgaac caattcttcg cgggcagaac aaaagcttct
     3601 gcacacgtct ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt gaatcgaacg
     3661 aaagacccgt gtgtaaagcc gcgtttccaa aatgtataaa accgagagca tctggccaat
     3721 gtgcatcagt tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa ctaaaggggg
     3781 gatctagatc ggggtaccat ggactacaaa gaccatgacg gtgattataa agatcatgat
     3841 atcgattaca aggatgacga tgacaaggat ccactagtca tgtctcacct gaccggcacc
     3901 cgcgtgagca ccttcaacga gagcttcctt aacgaaaatg agcacgactc gaatgcggtg
     3961 ctcatggaac tggacaaggg actgcggagc accaagcagg gaatccagtg cgaggcggtc
     4021 gtccgctttc cgcgcctctt cgagaagtat ccgtttccca ttctcatcaa ctcgtcgttc
     4081 attaagctgg ccgattactt tgtcagcgga tcgaatctgc tgcgcttctg ggtgctccgt
     4141 gtttgccagc agagcgagaa tcatctggac aagatcctca acattgacag ctttgtgcgc
     4201 tgtatctttg tggtgatgca ctccaacgat ccggtggcgc gtgctctcct cctgcgcacc
     4261 ttgggcgccg tttcccgtgt gattcccgag aagcaacagg ttcatcatgc cattcgacgt
     4321 gctctggata gccacgatac cgtcgaggtg gaggctgcca tctatgccag cagctgcttt
     4381 gccgcccagt caagttcctt tgccatcagc atgtgcgcca agatttcgga catgatcgaa
     4441 tcgcttcagg tgccggtgcc aatgaagctg ctcctgattc ccgtgttgcg acacatgcat
     4501 cacgaggcta ccacagcgtc actggttagc agactgtgca tggatctgct gcccaaatat
     4561 ccagcccaga gctttgtggt ggccatcata gacacgctca ctcaattgtc atcacgaacg
     4621 ctggtgggag tgcccggtca gctggatgtt ctgctggatt tcatgcagga cttaaggaca
     4681 ccagtacgca tacaggtgct gcgatctttc aacgagttgg ccggacgtca gagcgttcat
     4741 gcctggccca agccagccat caaagcgctc attgatcgtt ttgaattgtg taccaattcc
     4801 aaggagcagt tcctcttcct ctcaatcctg ctgaagttaa gcgagtgccc gcttacttgc
     4861 cagcagctgc ttcgcgagca tcgcgtagcc ctgctccgtt tgtgcatcca gtgcattagc
     4921 aagctggacg actacaccac cgccactcag gccatggcag ttttgtcggt tctagtagca
     4981 tttggtttaa aaaaaaaggg cagcggcgag caggtggacg atattcttca tatggtcaac
     5041 ctgcacatgg aaggattgct cctttgcacg gccaagcggt cggagtgcac ccgcgatctc
     5101 cgtagggtgc tgacatacgg tataaggatc actaaagcca acgctgaatt tggaacatcc
     5161 ttcattggga tcgtgaccaa tagcctggga gataagggtg cctatccgcc agccaatgcg
     5221 gaacttatgt gcgaggcact ggcgggtctg tgtgagcact ttcagctgcg taagtacgcc
     5281 ttttccaccg cagaagattt gatagtagac gaaaacgcga tggacaccga tgagttgccc
     5341 ccgcccgaga taaatcccat gctagctcgt ttgccgctca ttctgcacaa gctgaacacc
     5401 attatcgacc aggagaattg cgatcaacag ttgcgcagcg tggagatcct gagttctctg
     5461 gttctacaaa ccaccatggg ctgttacctg ccgcaaaagg ttgttcagtg ctttgagaaa
     5521 tgcctcggta ggcttaactg ctggacgctg tataggattg cacgcactgc tagtcgctat
     5581 ggtcaccact atgtggctgc tcacatttac accaaggtct cccagattgt gatcagcgat
     5641 catatgcact atttcctggt ggcactgtcg cagatctcgc aggcggagtg cattcttaac
     5701 tatggcttgg aatacgccta catgcgggac aattatgcac caaaggtggc accggaacct
     5761 ctaattccgc tgatgaagcg cttggaaatg gctagtaatc tctatcagca agcgctggcc
     5821 agcctgcgag ctggctcctc gccccagcat ccgtgcacct ttcagttgga gtatctgaag
     5881 ataagggcgc agtttctgca gacccttcac ctggctgtga ccgtaaagaa tgcccaggtg
     5941 attgtgccgc caccggcaat agctggaagt ttggcccaga actcgcgcga ctatctgcaa
     6001 aagtttggtc atgtcacgaa ccagctgcgc aagctggtga aggcactaaa ggcctgcgag
     6061 gagacttacg ctaggctata caaatcctct ttcgatgcgg atcacgtgac gctggagttt
     6121 ctggaggtgg ctgaatttca gtgcgctctt ttcgcccaca tcatcgagtc gatttgctac
     6181 gccacgcctc cggaaccgcc tgtatttcta accacaggcg atcacccgga gacgcgctac
     6241 tttgccgcaa gctgccagcg catggagcag atgcaaaaaa acctgccaca ggagccggcc
     6301 aatgccaaga ccattagtaa ccgacatttg gatgtcatca ttgcgcagat cgagatcata
     6361 acaaagacac cgctctgcct gccgcgttac ttcttccaga ttctgcagtc cacacagatc
     6421 aagctatcgg tgagtccgca gccaaggagc gccaccgaac ccgtgaatgt ccagtcgggc
     6481 agtaatctgg ttatcaaggt cgagggtgtc ctgcagcact ttagcaagca gaagaaacac
     6541 ttccgtcgcg tggagtccgt tcaattaagc ctcacctcac agctaatcac gccgccaccg
     6601 cgatcctcgc aagagctgcc caagcagggc gccaacgata ctgtgaccct caatcagatt
     6661 gtaaagcccc agcgcgattt cctctccggc agcttcctgt tgcccatctc caatggcggg
     6721 cacttccagg tgacgctgga aacattcgtg gtggacgaga atggcattac ctggtgcact
     6781 ggccccaaat cctcgatggt ggtgcgggtg ctggaggatc ctagcaagca gggcgctcca
     6841 gcgccgagca cttcccaggc agtgggacag acgaggaggt tttaactcga gtctagaggg
     6901 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     6961 cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc
     7021 taagatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     7081 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     7141 agctgcaata aacaagttct agggtcggtg ggcctcgggg gcgggtgcgg ggtcggcggg
     7201 gccgccccgg gtggcttcgg tcggagccat ggggtcgtgc gctcctttcg gtcgggcgct
     7261 gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc
     7321 cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt
     7381 tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc
     7441 cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg
     7501 tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt
     7561 gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca
     7621 ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg
     7681 cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg
     7741 tgacccgctc gatgtggcgg tccg
//
