LOCUS       sequence-416756-        5808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Drosophila mts protein with H241A 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 H241A
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..18)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(172..760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(252..271)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(931..1722,1723..1791))
                     /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     1554..1573
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1792..1896)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1964..1982
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2020..2042)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2142..2161
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    2275..2404
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     CDS             complement(2422..2439)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2449..2490)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(3452..3517)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(5578..5594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5578..5594)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5591..5613)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5602..5618
                     /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(5626..5632,5633..5650,5651..5656))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5671..5692
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
       61 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      121 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      181 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      241 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      301 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      361 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      421 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      481 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      541 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      601 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      661 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      721 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      781 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      841 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      901 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
      961 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1021 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1081 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1141 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1201 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1261 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1321 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1381 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1441 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1501 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1561 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1621 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1681 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     1741 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     1801 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1861 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1921 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     1981 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2041 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2101 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     2161 actgagagtg caccatatgg ctcacatggg cccatatggc tcacatgttc tttcctgcgt
     2221 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagcatcg atcccacgcg
     2281 cttgaaagga gtgtgtaaat ggacaatgtt gttacaataa aagttgaaat tattttataa
     2341 aatttattaa aaacatattt aaacataaca tattgtatta aaacatgaca atacaaacta
     2401 agatttagtc agataaactc aatggtgatg gtgatgatga ccggtacgcg tagaatcgag
     2461 accgaggaga gggttaggga taggcttacc ttcgaaccgc gggccctcta gattaaagga
     2521 aataatcggg tgttcttcgc gtaacatgag gctccccgcg cctgggtgct gggtcaaatt
     2581 gtaggaatga aaatttaagt gaatcatcca gttccataag agccgcttgg ttgccacagc
     2641 ggtagcaata gtttggcgct gagaatattg tgaccacatt gcgatcgtga caccagttgt
     2701 agccctccat caccagctga gcggcgcgcg acaccagtgt caggccgttt gtgttgttaa
     2761 aggtttccga aatatcctgg ccaaaggtgt aaccggcgcc acgaggcgag attccccagc
     2821 cacccctgtc atcgggatcg gaccagagca gatcgcacat gggaccctcg tgcggaacct
     2881 cctgcaagcg atcgagggcc cgaatgtgat ccagactgtc gatcgaggga ctgaggcctc
     2941 cgtgcaggca gaagatctgc ccgtcgacga gtgccgtcag tggcaagtag tcgaacagat
     3001 ccgtgaagta cttccaaacg ttggcattgc catacttgcg caggcactcg tcgtagaagc
     3061 cgtacacctg tgtgatctgg cgcgactcgt ggttaccgcg caggatggtg atgcgctcgc
     3121 gatagcgaac cttcagggcc accagaaggg tcacggtctc cacggagtag tatccacggt
     3181 ccacgtagtc gcccatgaac aggtagttgg tgtccggaga cttgccgcct atccggaaga
     3241 gctccatgag gtcgtgaaac tgaccgtgga catctccgca cactgtcacc gggcatttta
     3301 cctcctgcac attggactcc ttggagagaa tctccttggc cttgtcgcag agggtgcgaa
     3361 cttgtgtctc tgtcaactga ttgcattcgt tcaactgctc aatccattga tcaagatctt
     3421 ttgttgttgc tttatcctcg actagtggat ccttgtcatc gtcatccttg taatcgatat
     3481 catgatcttt ataatcaccg tcatggtctt tgtagtccat aagcttcctc tagctagagt
     3541 cgacctgcag tttggattat tctgcgggca gaaaatagag atgtggaaaa ttagtacgaa
     3601 atcaaatgag tttcgttgaa attacaaaac tattgaaact aacttcctgg ctggggaata
     3661 aaaatgggaa acttatttat cgacgccaac tttgttgaga aacccctatt aaccctctac
     3721 gaatattgga acaaaggaaa gcgaagaaac agggaacaaa gggtagttga gaaacctttg
     3781 ttccgttgct tcgtcatcgt tttcgtgttg cgagtgtgtg catgtatata tacacagctg
     3841 aaatgcatgc atacacatta ttttgtgtgt atatggtgac gtcacaacta caagcaataa
     3901 gaaattttca gacgtggctt tgtttaagca acctactcta tttcagctaa aaataagtgg
     3961 atttcgttgg taaaatactt caattaacta aacaactaac taataacatg cacacaaatg
     4021 ctcgatcgag tgcgttcgtg atttctcgaa ttttcaaatg cgtcactgcg aatttcacaa
     4081 tttgccaata aatcttggcg aaaatcaaca cgcaagtttt atttatagat ttgtttgcgt
     4141 tttgatgcca attcattggg aaaacaagat gcgtggctgc caatttctta ttttgtaatt
     4201 acgtagagcg ttgaataaaa aaaaatggcc gaacaaagac cttgaaatgc agtttttctt
     4261 gaaattactc aacgtcttgt tgctcttatt actaattggt aacagcgagt taaaaactta
     4321 cgtttcttgt gactttcgag aatgttcttt taattgtact ttaatcacca acaattaagt
     4381 ataaattttt cgctgattgc gctttacttt ctacttgtac ttgctgctgc aaatgtcaat
     4441 tggttttgaa ggcgaccgtt cgcgaacgct gtttatatac cttcggtgtc cgttgaaaat
     4501 cactaaaaaa taccgtagtg ttcgtaacac tttagtacag agaaaaaaaa ttgtgccgaa
     4561 atgtttttga tacgtacgaa taccttgtat taaaattttt tatgatttct gtgtatcact
     4621 ttttttttgt gtttttcgtt taaactcacc acagtacaaa acaataaaat atttttaaga
     4681 caatttcaaa ttgagacctt tctcgtactg acttgaccgg ctgaatgagg atttctacct
     4741 agacgaccta cttcttacca tgacattgaa tgcaatgcca cctttgatct aaacttacaa
     4801 aagtccaagg cttgttagga ttggtgtttt ggaatttaag acccttgtta gtttgctttt
     4861 gaaatagcac tgtcttctct accggctata attttgaaac tcgcagcttg actggaaatt
     4921 taaaaagtaa ttctgtgtag gtaaagggtg ttttaaaagt gtgatgtgtt gagcgttgcg
     4981 gcaacgactg ctatttatgt atatattttc aaaacttatt gtttttgaag tgttttaaat
     5041 ggagctatct ggcaacgctg cgcataatct tacacaagct agcttttctt aatccatttt
     5101 taagtgaaat ttgtttatac tctttcggca aataattgtt aaatcgcttt aagtgggctt
     5161 acatctggat aagtaatgaa aacctgcata ttataatatt aaaacatata atccactgtg
     5221 ctttccccgt gtgtggccat atacctaaaa aagtttattt tcgcagagcc ccgcacggtc
     5281 acactacggt tcggcgattt tcgattttgg acagtactga ttgcaagcgc accgaaagca
     5341 aaatggagct ggagattttg aacgcgaaga acagcaagcc gtacggcaag gtgaaggtgc
     5401 cctccggcgc cacgcccatc ggcgatctgc gcgccctaat tcacaagacc ctgaagcaga
     5461 ccccacacgc gaatcgccag tcgcttcgtc tggaactgaa gggcaaaagc ctgaaagata
     5521 cggacacatt ggaatctctg tcgctgcgtt ccggcgacaa agcttggcgt aatcatggtc
     5581 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5641 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5701 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5761 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttc
//
