LOCUS       sequence-450117-        5460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For purification of N-terminally His-tagged TRPT1 from bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5460)
  AUTHORS   Weixler L, Feijs KLH, Zaja R
  TITLE     ADP-ribosylation of RNA in mammalian cells is mediated by TRPT1 and 
            multiple PARPs.
  JOURNAL   Nucleic Acids Res. 2022 Sep 9;50(16):9426-9441. doi: 
            10.1093/nar/gkac711.
  PUBMED    36018800
REFERENCE   2  (bases 1 to 5460)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2022-09-9- 9"; volume: "50"; issue: 
            "16"; pages: "9426-9441"
COMMENT     Sequence Label: pDEST17-hTRPT1-iso1
FEATURES             Location/Qualifiers
     source          1..5460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        23..41
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             73..95
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             115..132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    142..166
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(931..955)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1008..1026)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1022..1069
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1435..1440,1441..1457,1458..1463))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1490..1508)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1576..1680
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1681..1749,1750..2541)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1899..1918)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2712..3300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3201..3220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3454..3471
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3486..3625
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3551..3570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3711..3733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3727..3918)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5380..5399
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgagatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
       61 tctagaaata attttgttta actttaagaa ggagatatac atatgtcgta ctaccatcac
      121 catcaccatc acctcgaatc aacaagtttg tacaaaaaag caggcttcat gaacttctct
      181 ggaggaggga ggcaggaagc agcagggtcc aggggtagaa gggctcccag accccgagaa
      241 caggaccgag acgtgcagct gtccaaggct ctgtcctatg ccctgcgcca tggggccttg
      301 aagctggggc ttcccatggg agctgatggc ttcgtgcccc tgggcaccct cctgcagttg
      361 ccccagttcc gcggcttctc tgctgaagat gtgcagcgcg tggtggacac caataggaag
      421 cagcggttcg ccctgcagct gggggatccc agcactggcc ttctcatccg ggccaaccag
      481 ggccattccc tgcaggtacc taagttggag ctgatgcccc tggagacacc gcaggccctg
      541 cccccgatgc tagtccatgg tacattctgg aagcactggc catccatcct actcaaaggc
      601 ctgtcctgcc agggaaggac gcacattcac ctggccccag gactgcctgg agaccccggt
      661 atcatcagtg gcatgcggtc ccattgtgaa atagctgtgt tcatcgatgg acccctggct
      721 ctggcagatg gaataccctt cttccgctct gccaatgggg tgattctgac tccagggaat
      781 actgatggct tcctccttcc caagtacttc aaggaggccc tgcagctacg ccctacccga
      841 aagccccttt ccttggctgg tgatgaagag acagagtgtc agagtagccc caagcacagc
      901 tccagagaaa ggaggaggat ccaacaataa acccagcttt cttgtacaaa gtggttgatt
      961 cgaggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     1021 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     1081 aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg
     1141 ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga
     1201 gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac
     1261 tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca
     1321 agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag
     1381 atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa
     1441 gcttatcgat gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata
     1501 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     1561 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1621 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1681 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1741 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1801 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1861 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1921 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1981 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2041 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2101 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2161 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2221 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2281 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2341 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2401 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2461 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2521 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2581 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2641 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2701 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2761 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2821 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     2881 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2941 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3001 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3061 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3121 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3181 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3241 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3301 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3361 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3421 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3481 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt
     3541 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     3601 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     3661 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3721 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     3781 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     3841 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     3901 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     3961 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     4021 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     4081 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     4141 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     4201 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     4261 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     4321 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     4381 ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga
     4441 cgcgatggat atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg
     4501 attggctcca attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag
     4561 gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg
     4621 gcggcgccta caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc
     4681 gtgacgatca gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga
     4741 agctgtccct gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc
     4801 ccgatgccgc cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg
     4861 aacgccagca agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc
     4921 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     4981 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     5041 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     5101 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     5161 gctctcaagg gcatcggtcg atcgacgctc tcccttatgc gactcctgca ttaggaagca
     5221 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     5281 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     5341 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     5401 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
//
