LOCUS       sequence-451135-        6259 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 6259)
  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 6259)
  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-LAP3
FEATURES             Location/Qualifiers
     source          1..6259
                     /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(1728..1752)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1805..1823)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1819..1866
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2232..2237,2238..2254,2255..2260))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2287..2305)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2373..2477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2478..2546,2547..3338)
                     /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(2696..2715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3509..4097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3998..4017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4251..4268
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4283..4422
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4348..4367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4508..4530
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4524..4715)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6177..6196
                     /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 ggcttcatgt tcttgctgcc
      181 tcttccggct gcggggcgag tagtcgtccg acgtctggcc gtgagacgtt tcgggagccg
      241 gagtctctcc accgcagaca tgacgaaggg ccttgtttta ggaatctatt ccaaagaaaa
      301 agaagatgat gtgccacagt tcacaagtgc aggagagaat tttgataaat tgttagctgg
      361 aaagctgaga gagactttga acatatctgg accacctctg aaggcaggga agactcgaac
      421 cttttatggt ctgcatcagg acttccccag cgtggtgcta gttggcctcg gcaaaaaggc
      481 agctggaatc gacgaacagg aaaactggca tgaaggcaaa gaaaacatca gagctgctgt
      541 tgcagcgggg tgcaggcaga ttcaagacct ggagctctcg tctgtggagg tggatccctg
      601 tggagacgct caggctgctg cggagggagc ggtgcttggt ctctatgaat acgatgacct
      661 aaagcaaaaa aagaagatgg ctgtgtcggc aaagctctat ggaagtgggg atcaggaggc
      721 ctggcagaaa ggagtcctgt ttgcttctgg gcagaacttg gcacgccaat tgatggagac
      781 gccagccaat gagatgacgc caaccagatt tgctgaaatt attgagaaga atctcaaaag
      841 tgctagtagt aaaaccgagg tccatatcag acccaagtct tggattgagg aacaggcaat
      901 gggatcattc ctcagtgtgg ccaaaggatc tgacgagccc ccagtcttct tggaaattca
      961 ctacaaaggc agccccaatg caaacgaacc acccctggtg tttgttggga aaggaattac
     1021 ctttgacagt ggtggtatct ccatcaaggc ttctgcaaat atggacctca tgagggctga
     1081 catgggagga gctgcaacta tatgctcagc catcgtgtct gctgcaaagc ttaatttgcc
     1141 cattaatatt ataggtctgg cccctctttg tgaaaatatg cccagcggca aggccaacaa
     1201 gccgggggat gttgttagag ccaaaaacgg gaagaccatc caggttgata acactgatgc
     1261 tgaggggagg ctcatactgg ctgatgcgct ctgttacgca cacacgttta acccgaaggt
     1321 catcctcaat gccgccacct taacaggtgc catggatgta gctttgggat caggtgccac
     1381 tggggtcttt accaattcat cctggctctg gaacaaactc ttcgaggcca gcattgaaac
     1441 aggggaccgt gtctggagga tgcctctctt cgaacattat acaagacagg ttgtagattg
     1501 ccagcttgct gatgttaaca acattggaaa atacagatct gcaggagcat gtacagctgc
     1561 agcattcctg aaagaattcg taactcatcc taagtgggca catttagaca tagcaggcgt
     1621 gatgaccaac aaagatgaag ttccctatct acggaaaggc atgactggga ggcccacaag
     1681 gactctcatt gagttcttac ttcgtttcag tcaagacaat gcttagtacc cagctttctt
     1741 gtacaaagtg gttgattcga ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1801 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1861 tttttgctga aaggaggaac tatatccgga tatccacagg acgggtgtgg tcgccatgat
     1921 cgcgtagtcg atagtggctc caagtagcga agcgagcagg actgggcggc ggccaaagcg
     1981 gtcggacagt gctccgagaa cgggtgcgca tagaaattgc atcaacgcat atagcgctag
     2041 cagcacgcca tagtgactgg cgatgctgtc ggaatggacg atatcccgca agaggcccgg
     2101 cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac
     2161 gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga
     2221 taaactaccg cattaaagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac
     2281 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     2341 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2401 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2461 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2521 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2581 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2641 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2701 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     2761 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2821 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2881 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2941 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3001 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3061 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3121 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3181 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3241 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3301 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3361 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3421 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3481 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3541 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3601 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3661 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3721 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3781 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3841 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3901 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3961 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4021 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4081 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4141 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4201 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4261 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4321 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4381 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     4441 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4501 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     4561 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     4621 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     4681 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     4741 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     4801 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     4861 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     4921 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     4981 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     5041 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     5101 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     5161 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg
     5221 ctgctggaga tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca
     5281 gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc
     5341 cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga
     5401 ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg
     5461 aggcggcata aatcgccgtg acgatcagcg gtccaatgat cgaagttagg ctggtaagag
     5521 ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg
     5581 cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca
     5641 tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg
     5701 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     5761 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     5821 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     5881 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     5941 agctgactgg gttgaaggct ctcaagggca tcggtcgatc gacgctctcc cttatgcgac
     6001 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     6061 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     6121 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     6181 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     6241 cgtccggcgt agaggatcg
//
