LOCUS       sequence-416777-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-tagged Drosophila IntS13 from inducible MtnA 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6843)
  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 6843)
  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-IntS13 puro
FEATURES             Location/Qualifiers
     source          1..6843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     88..107
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(721..737)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(721..737)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(734..756)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    745..761
                     /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(769..775,776..793,794..799))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    814..835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(952..969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1123..1711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1203..1222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1882..2673,2674..2742))
                     /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     2505..2524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2743..2847)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2915..2933
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2971..2993)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3093..3112
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3306..3328
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3320..3337
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3321..3337
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3343..3769
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3794..3859
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5985..6026
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6036..6053
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6076..6093)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6095..6229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6119..6138)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6173..6192
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6351..6843,1..107))
                     /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(6434..6454)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggcccggggg
       61 acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatgga aggtcgtctc
      121 cttgtgaggg gtcaggggcg tgggtcaggg gatggtggcg gcaccggtcg tggcggccga
      181 cctgcaggca tgcaagctat ccaattcctg cagcccgggg gatctgttgt aatttataat
      241 ttatatttcc tttcttaata aataaataaa tagtcaagtt tatgtttgag ttttatgatt
      301 tatattttaa gttatttcaa ctgcaacacc agcaccacga cctacttaca gcaaaaaacg
      361 tacaagaagg aaagaaggaa taaaaagagt ggtattctct tacaatatgt tttatggcat
      421 aaaaggtgtg gccattcata tcaaatataa agtagtgttg tttaacgtta cttttgtagg
      481 ttgaatagta tattccaaca gatgatgagg ggttcccaat cctaaaccca tttgccgttc
      541 ccagaagcat gaaaccacca cgcacccgat cctctaggac aacaacaatt gcattcattt
      601 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
      661 atgtggtatg gctgattatg atcagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
      721 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
      781 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      841 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      901 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     1321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1561 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1681 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1741 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1801 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1861 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1921 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1981 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2041 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2101 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2161 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2221 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2281 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2341 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2401 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2461 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2521 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2581 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     2641 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2701 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2761 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2821 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     2881 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2941 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     3001 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     3061 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     3121 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     3181 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     3241 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     3301 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg
     3361 atgtggtgcc cgatgtgact agctctttgc tgcaggccgt cctatcctct ggttccgata
     3421 agagacccag aactccggcc ccccaccgcc caccgccacc cccatacata tgtggtacgc
     3481 aagtaagagt gcctgcgcat gccccatgtg ccccaccaag agctttgcat cccatacaag
     3541 tccccaaagt ggagaaccga accaattctt cgcgggcaga acaaaagctt ctgcacacgt
     3601 ctccactcga atttggagcc ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc
     3661 gtgtgtaaag ccgcgtttcc aaaatgtata aaaccgagag catctggcca atgtgcatca
     3721 gttgtggtca gcagcaaaat caagtgaatc atctcagtgc aactaaaggg gggatctaga
     3781 tcggggtacc atggactaca aagaccatga cggtgattat aaagatcatg atatcgatta
     3841 caaggatgac gatgacaagg atccactagt ccagtgtggt ggaattctgt tcgaacgcaa
     3901 ccagaagacc atctttgtgc tggaccacac ccgatacttt agcatcgcca gcgaggagta
     3961 catctcgatg gacttcctaa agggaaaacc atctgcagac ggcggcgcaa caggagcggc
     4021 gggaaatgca accggcagcg gtggatccca attcagcaag agcctgtgga catgcgcctg
     4081 cgaatcctcc atcgagtact gccgcgtggt ctgggatctc tttcccggca aaaagcatgt
     4141 gcggtttatt gtctcggaca cggcggcgca tatcgtgaac acctggaggc ccagcacgca
     4201 aaatatggcg cacgtgatga acgcaatgct aattgtgggc gtgccgtcgc gcaacgtacc
     4261 cacatcttct gactactcgg taatccatgg cctgcgggcc gccatcgaag cgctggcgga
     4321 gcccaccgat gagcagttgg cggcaatggc cgactttggg accgacgaac tgccgcgcat
     4381 tcccaacaag ggtcgcgtca tctgcataac ttcagcgcgt gacaatacca gcatgaaaag
     4441 cctggaggac atctttaaca ccgtgctggt gcagcagaac accctagcag ctccgccctc
     4501 caagaaggga ctggtcatcg accattgcca cctagttatc ctcaacattg tgccgctggg
     4561 cgttgaatcg ttggttacca atcgcagcct gctcaagata tcgccgctgc ttgatgtcga
     4621 gatccacaca gtcagtgccc cggatatttc ctataagctg acgcacctca tactgaacca
     4681 ttacgaccta gctagtacca cggtgactaa catacccatg aaagaggagc agaacgccaa
     4741 ctccagcgcc aactacgatg tggaaatcct gcacagccga agggcccact ccatcacctg
     4801 tggcccagat ttcagtctac ccacgagcat caagcaaggt gccacatacg aaacagtgac
     4861 tcttaagtgg tgcacaccgc gcggttgcgg ctctgcagat ctacagccct gtctgggtca
     4921 gttcctcgtg acaccggtgg acgtcacctc gcggcccagt tcttgcttga ttaactttct
     4981 gcttaacgga cgttcagtct tgctggaaat gccgcgcaag accggctcaa aggccaccag
     5041 ccatatgctt tctgcacgcg gtggcgagat ctttgtgcac tccctgtgca tcacgcgctc
     5101 ctgcatggat gaggctccat caattactga tggccccgga ggacgtgtct cggactatcg
     5161 caccgccgaa cttggtcaac tgatcaagat gtcgcgcgtg gttccgctca aggttaagga
     5221 tccatctgct ccgcccttga cgcgccggtt gcctcgctac ttccccctga ccaccagctc
     5281 ttcgattctg ttccatctac agcgacacat cagttggctt ccgcattttc tccatctttt
     5341 agtgaaggaa gacatggata agcaggacga ggtgcggtgc cagcagcaca ttcacgagct
     5401 gtacaaaagc gcctcgcgtg gcgatgtcct accatttacc cacaccaatg gagcaagact
     5461 aaagctttcc aaggccaaag accagtaccg cttgctgtac agggaactgg agcaattaat
     5521 tcaactaaat gccaccacaa tgcaccataa aaatctgctg gaaagcctac agagtttgcg
     5581 agccgcatac ggcgatgccc cgctgaagtc ggaacctgga gcaagtctcc tgcgcaccta
     5641 cacggaatcc ccactctctc ccgaacggct ggagcccatc agtagcgtcg gtgctagtgg
     5701 cagcagcagc tccaatagcc ttctcaaggc gagcaagcgt cgcatgtcta gttgtgggca
     5761 aagatccctg ctagatatta tatcctcagc agagcgaagc cagtccaaca aacgattgga
     5821 tttctcggga cgcctttgca ctccgttggg tcaagtggcc aagctgtatc cggattttgg
     5881 taccaaggac aaagacacag ttacgacggg ggccagtatt acgcccaatg ttaaggagga
     5941 atccgtacgt agttaactcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta
     6001 accctctcct cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt
     6061 aaacccgctg atcagcctcg actgtgcctt ctaagatcca gacatgataa gatacattga
     6121 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     6181 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttc tagggtcggt
     6241 gggcctcggg ggcgggtgcg gggtcggcgg ggccgccccg ggtggcttcg gtcggagcca
     6301 tggggtcgtg cgctcctttc ggtcgggcgc tgcgggtcgt ggggcgggcg tcaggcaccg
     6361 ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg
     6421 gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg
     6481 ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc
     6541 ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc
     6601 cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac
     6661 tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg
     6721 gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg
     6781 acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccgggtcg
     6841 acg
//
