LOCUS       sequence-451164-        5356 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For protein purification from Ecoli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5356)
  AUTHORS   Weixler L, Zaja R, Ikenga NJ, Mohan G, Aydin G, Wijngaarden S, 
            Filippov DV, Luscher B, Feijs-Zaja KLH
  TITLE     Family-wide analysis of human macrodomains reveals novel activities
  JOURNAL   ResearchSquare. 2024 Aug 8. doi: 10.21203/rs.3.rs-4644671/v1
REFERENCE   2  (bases 1 to 5356)
  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: 
            "ResearchSquare. 2024 Aug 8. doi: 10.21203/rs.3.rs-4644671/v1"
COMMENT     Sequence Label: pDEST17-NUDT5
FEATURES             Location/Qualifiers
     source          1..5356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             113..130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    140..164
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(825..849)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(902..920)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      916..963
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1329..1334,1335..1351,1352..1357))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1384..1402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1470..1574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1575..1643,1644..2435)
                     /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(1793..1812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2606..3194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3095..3114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3348..3365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3380..3519
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3445..3464)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3605..3627
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3621..3812)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5274..5293
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacat atgtcgtact accatcacca
      121 tcaccatcac ctcgaatcaa caagtttgta caaaaaagca ggcttcgaga gccaagagcc
      181 aaccgaaagc tcccagaacg ggaaacagta cattatttcg gaagagttaa tcagcgaagg
      241 taagtgggtc aaacttgaaa agaccaccta catggatcct actggcaaga cacgtacatg
      301 ggaatctgtt aaacgtacta cacgtaagga acagaccgct gatggagttg ccgtaattcc
      361 agttcttcag cgtactcttc actacgaatg catcgttctg gtaaagcagt ttcgcccacc
      421 gatgggaggt tattgtattg agtttcccgc tggtcttatc gatgacggtg agacaccaga
      481 ggctgcggca ttgcgtgagc ttgaggaaga aaccggttat aagggcgaca tcgcggagtg
      541 ttcaccagct gtttgtatgg acccgggttt atcaaactgt actattcaca ttgtaactgt
      601 cacgatcaat ggtgacgatg cagagaacgc tcgtccaaaa ccgaaacctg gcgatggcga
      661 gtttgtagag gtaatctcct tgccaaagaa cgacctttta cagcgcttgg acgcccttgt
      721 tgctgaggaa catttgacag ttgatgcgcg tgtttactct tacgccttgg cgcttaaaca
      781 cgccaacgct aagccgtttg aagtaccctt cttaaagttt taatacccag ctttcttgta
      841 caaagtggtt gattcgaggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
      901 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      961 ttgctgaaag gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc
     1021 gtagtcgata gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc
     1081 ggacagtgct ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag
     1141 cacgccatag tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag
     1201 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     1261 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     1321 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
     1381 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1441 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1501 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1561 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1621 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1681 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1741 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1801 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1861 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1921 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1981 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2041 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2101 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2161 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2221 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2281 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2341 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2401 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2461 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2521 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3301 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3361 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3421 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3481 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3541 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3601 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3661 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3721 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3781 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3841 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     3901 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     3961 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     4021 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     4081 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     4141 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     4201 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4261 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     4321 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     4381 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     4441 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     4501 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     4561 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     4621 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     4681 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     4741 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     4801 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     4861 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     4921 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     4981 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     5041 tgactgggtt gaaggctctc aagggcatcg gtcgatcgac gctctccctt atgcgactcc
     5101 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     5161 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     5221 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     5281 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     5341 ccggcgtaga ggatcg
//
