LOCUS       sequence-416759-        5808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Drosophila mts protein with H59Q mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5808)
  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 5808)
  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: pUb FLAG-Mts H59Q
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     16..34
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(72..94)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     194..213
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    327..456
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     CDS             complement(474..491)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(501..542)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(1504..1569)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3630..3646)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3630..3646)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3643..3665)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3654..3670
                     /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(3678..3684,3685..3702,3703..3708))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3723..3744
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3861..3878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4032..4620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4112..4131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4791..5582,5583..5651))
                     /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     5414..5433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5652..5756)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
       61 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
      121 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
      181 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat ggctcacatg
      241 ggcccatatg gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      301 taccgccttt gagtgagcat cgatcccacg cgcttgaaag gagtgtgtaa atggacaatg
      361 ttgttacaat aaaagttgaa attattttat aaaatttatt aaaaacatat ttaaacataa
      421 catattgtat taaaacatga caatacaaac taagatttag tcagataaac tcaatggtga
      481 tggtgatgat gaccggtacg cgtagaatcg agaccgagga gagggttagg gataggctta
      541 ccttcgaacc gcgggccctc tagattaaag gaaataatcg ggtgttcttc gcgtaacatg
      601 aggctccccg cgcctgggtg ctgggtcaaa ttgtaggaat gaaaatttaa gtgaatcatc
      661 cagttccata agagccgctt ggttgccaca gcggtagcaa tagtttggcg ctgagaatat
      721 tgtgaccaca ttgcgatcgt gacaccagtt gtagccctcc atcaccagct gatgggcgcg
      781 cgacaccagt gtcaggccgt ttgtgttgtt aaaggtttcc gaaatatcct ggccaaaggt
      841 gtaaccggcg ccacgaggcg agattcccca gccacccctg tcatcgggat cggaccagag
      901 cagatcgcac atgggaccct cgtgcggaac ctcctgcaag cgatcgaggg cccgaatgtg
      961 atccagactg tcgatcgagg gactgaggcc tccgtgcagg cagaagatct gcccgtcgac
     1021 gagtgccgtc agtggcaagt agtcgaacag atccgtgaag tacttccaaa cgttggcatt
     1081 gccatacttg cgcaggcact cgtcgtagaa gccgtacacc tgtgtgatct ggcgcgactc
     1141 gtggttaccg cgcaggatgg tgatgcgctc gcgatagcga accttcaggg ccaccagaag
     1201 ggtcacggtc tccacggagt agtatccacg gtccacgtag tcgcccatga acaggtagtt
     1261 ggtgtccgga gacttgccgc ctatccggaa gagctccatg aggtcgtgaa actgaccctg
     1321 gacatctccg cacactgtca ccgggcattt tacctcctgc acattggact ccttggagag
     1381 aatctccttg gccttgtcgc agagggtgcg aacttgtgtc tctgtcaact gattgcattc
     1441 gttcaactgc tcaatccatt gatcaagatc ttttgttgtt gctttatcct cgactagtgg
     1501 atccttgtca tcgtcatcct tgtaatcgat atcatgatct ttataatcac cgtcatggtc
     1561 tttgtagtcc ataagcttcc tctagctaga gtcgacctgc agtttggatt attctgcggg
     1621 cagaaaatag agatgtggaa aattagtacg aaatcaaatg agtttcgttg aaattacaaa
     1681 actattgaaa ctaacttcct ggctggggaa taaaaatggg aaacttattt atcgacgcca
     1741 actttgttga gaaaccccta ttaaccctct acgaatattg gaacaaagga aagcgaagaa
     1801 acagggaaca aagggtagtt gagaaacctt tgttccgttg cttcgtcatc gttttcgtgt
     1861 tgcgagtgtg tgcatgtata tatacacagc tgaaatgcat gcatacacat tattttgtgt
     1921 gtatatggtg acgtcacaac tacaagcaat aagaaatttt cagacgtggc tttgtttaag
     1981 caacctactc tatttcagct aaaaataagt ggatttcgtt ggtaaaatac ttcaattaac
     2041 taaacaacta actaataaca tgcacacaaa tgctcgatcg agtgcgttcg tgatttctcg
     2101 aattttcaaa tgcgtcactg cgaatttcac aatttgccaa taaatcttgg cgaaaatcaa
     2161 cacgcaagtt ttatttatag atttgtttgc gttttgatgc caattcattg ggaaaacaag
     2221 atgcgtggct gccaatttct tattttgtaa ttacgtagag cgttgaataa aaaaaaatgg
     2281 ccgaacaaag accttgaaat gcagtttttc ttgaaattac tcaacgtctt gttgctctta
     2341 ttactaattg gtaacagcga gttaaaaact tacgtttctt gtgactttcg agaatgttct
     2401 tttaattgta ctttaatcac caacaattaa gtataaattt ttcgctgatt gcgctttact
     2461 ttctacttgt acttgctgct gcaaatgtca attggttttg aaggcgaccg ttcgcgaacg
     2521 ctgtttatat accttcggtg tccgttgaaa atcactaaaa aataccgtag tgttcgtaac
     2581 actttagtac agagaaaaaa aattgtgccg aaatgttttt gatacgtacg aataccttgt
     2641 attaaaattt tttatgattt ctgtgtatca cttttttttt gtgtttttcg tttaaactca
     2701 ccacagtaca aaacaataaa atatttttaa gacaatttca aattgagacc tttctcgtac
     2761 tgacttgacc ggctgaatga ggatttctac ctagacgacc tacttcttac catgacattg
     2821 aatgcaatgc cacctttgat ctaaacttac aaaagtccaa ggcttgttag gattggtgtt
     2881 ttggaattta agacccttgt tagtttgctt ttgaaatagc actgtcttct ctaccggcta
     2941 taattttgaa actcgcagct tgactggaaa tttaaaaagt aattctgtgt aggtaaaggg
     3001 tgttttaaaa gtgtgatgtg ttgagcgttg cggcaacgac tgctatttat gtatatattt
     3061 tcaaaactta ttgtttttga agtgttttaa atggagctat ctggcaacgc tgcgcataat
     3121 cttacacaag ctagcttttc ttaatccatt tttaagtgaa atttgtttat actctttcgg
     3181 caaataattg ttaaatcgct ttaagtgggc ttacatctgg ataagtaatg aaaacctgca
     3241 tattataata ttaaaacata taatccactg tgctttcccc gtgtgtggcc atatacctaa
     3301 aaaagtttat tttcgcagag ccccgcacgg tcacactacg gttcggcgat tttcgatttt
     3361 ggacagtact gattgcaagc gcaccgaaag caaaatggag ctggagattt tgaacgcgaa
     3421 gaacagcaag ccgtacggca aggtgaaggt gccctccggc gccacgccca tcggcgatct
     3481 gcgcgcccta attcacaaga ccctgaagca gaccccacac gcgaatcgcc agtcgcttcg
     3541 tctggaactg aagggcaaaa gcctgaaaga tacggacaca ttggaatctc tgtcgctgcg
     3601 ttccggcgac aaagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3661 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3721 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3781 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3841 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3901 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3961 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4021 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4081 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4141 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4201 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4261 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4321 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4381 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4441 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4501 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4561 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4621 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4681 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4741 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4801 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4861 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4921 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4981 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5041 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5101 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5161 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5221 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5281 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5341 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5401 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5461 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5521 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5581 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5641 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5701 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5761 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatg
//
