LOCUS       sequence-384211-        5775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Drosophila IntS12 from inducible MtnA 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5775)
  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 5775)
  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-IntS12 puro
FEATURES             Location/Qualifiers
     source          1..5775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      909..1497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1398..1417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1651..1668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1785..1806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1821..1826,1827..1844,1845..1851)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1859..1875
                     /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)."
     primer_bind     1864..1886
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1883..1899
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1883..1899
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2513..3112
                     /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(2513..2532)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3009..3029
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3234..3368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3271..3290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3325..3344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3370..3387
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3410..3427)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3437..3478)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(4536..4601)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(4626..5052)
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     primer_bind     complement(5058..5075)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5058..5074)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5067..5089)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5283..5302)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5402..5424
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5462..5480)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5548..5652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5653..5721,5722..5775,1..738)
                     /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"
ORIGIN
        1 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
       61 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      121 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      421 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1021 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1681 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     1741 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     1801 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     1861 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca
     1921 tgcctgcagg tcgactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1981 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2041 tcctagagga tcgggtgcgt ggtggtttca tgcttctggg aacggcaaat gggtttagga
     2101 ttgggaaccc ctcatcatct gttggaatat actattcaac ctacaaaagt aacgttaaac
     2161 aacactactt tatatttgat atgaatggcc acacctttta tgccataaaa catattgtaa
     2221 gagaatacca ctctttttat tccttctttc cttcttgtac gttttttgct gtaagtaggt
     2281 cgtggtgctg gtgttgcagt tgaaataact taaaatataa atcataaaac tcaaacataa
     2341 acttgactat ttatttattt attaagaaag gaaatataaa ttataaatta caacagatcc
     2401 cccgggctgc aggaattgga tagcttgcat gcctgcaggt cggccgccac gaccggtgcc
     2461 gccaccatcc cctgacccac gcccctgacc cctcacaagg agacgacctt ccatgaccga
     2521 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     2581 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     2641 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     2701 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     2761 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     2821 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     2881 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     2941 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     3001 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     3061 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     3121 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctccg accgaagcca
     3181 cccggggcgg ccccgccgac cccgcacccg cccccgaggc ccaccgaccc tagaacttgt
     3241 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3301 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3361 tctggatctt agaaggcaca gtcgaggctg atcagcgggt ttaaactcaa tggtgatggt
     3421 gatgatgacc ggtacgcgta gaatcgagac cgaggagagg gttagggata ggcttacctt
     3481 cgaaccgcgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattct
     3541 tactgcttgg atctgcgctt gctgctgctg ccgctttcgt gatgcttgga ggaactgctc
     3601 ttggagctgg atttggtgct ggagccgctg cgtcgcgatg aggtgccact gctgttgtgc
     3661 ttctccattc cgccgatggc gctaagagaa gacgccgccg tggacttaga ggatcgttct
     3721 tccttgtgcg atttggacga cgacgaagag gaggacttgt ggccatgctt gctgctgcta
     3781 ctgctgctgg cattcgtgga actgctgggc tcgggccggt agaagggtga cgaggagttg
     3841 gaagagttcg atgagcgcga cgacgcgacc gacgatttgg ctttgctggt caacggcatg
     3901 ggttcgtcgg ctatgaagac agttggcgtt acggccgctg cagaggatgt tgtcctcccg
     3961 ctgctcgttg gcttgttgca gcacgtgtcg cactgccagt tctgctcctg gtcatcggcc
     4021 gcctcctcct tggttatggg cggcttgtgg cactcctgat ggtacatggc accgcacttg
     4081 gagcactcaa tcagccgatt ggtggccgta aacaccattt ctccacagac acaacagttc
     4141 agatctccga aatctcccgt atcgcccgta ttcacgatgc cgaccgcgga caggccatcg
     4201 tcgctgtcgg cgatggtatc gacgctgtcc gacggttctt tatcggggga attagtcagg
     4261 ttaataattt catcggcgtt aatcggttgt tggggcgggg tcgcggcgcg gcccgagaaa
     4321 ttggcctcgt cttcgagcat gcggggggtc atattattcg tcagggactt ttcggggccg
     4381 aagcgggctt tcagggcttc atccagcagg aggcggagct ccgcggcgct cgtcgggttg
     4441 gagctatgga gcagtttaat cgcttttttg aggaccgggt cgacctcctg cgcggcggcg
     4501 gcggccgccg cgatgttggc cgcgactagt ggatccttgt catcgtcatc cttgtaatcg
     4561 atatcatgat ctttataatc accgtcatgg tctttgtagt ccatggtacc ccgatctaga
     4621 tccccccttt agttgcactg agatgattca cttgattttg ctgctgacca caactgatgc
     4681 acattggcca gatgctctcg gttttataca ttttggaaac gcggctttac acacgggtct
     4741 ttcgttcgat tcacctcttt tgcacacgcc ggccggctcc aaattcgagt ggagacgtgt
     4801 gcagaagctt ttgttctgcc cgcgaagaat tggttcggtt ctccactttg gggacttgta
     4861 tgggatgcaa agctcttggt ggggcacatg gggcatgcgc aggcactctt acttgcgtac
     4921 cacatatgta tgggggtggc ggtgggcggt ggggggccgg agttctgggt ctcttatcgg
     4981 aaccagagga taggacggcc tgcagcaaag agctagtcac atcgggcacc acatcctgtc
     5041 ctgcaacgaa ttaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     5101 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5161 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     5221 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     5281 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5341 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5401 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5461 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5521 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5581 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5641 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5701 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5761 cagttgggtg cacga
//
