LOCUS       sequence-134677-        4444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of human DNA Polymerase beta (POLB).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4444)
  AUTHORS   Schuermann D, Scheidegger SP, Weber AR, Bjoras M, Leumann CJ, Schar 
            P
  TITLE     3CAPS - a structural AP-site analogue as a tool to investigate DNA 
            base excision repair.
  JOURNAL   Nucleic Acids Res. 2016 Jan 4. pii: gkv1520.
  PUBMED    26733580
REFERENCE   2  (bases 1 to 4444)
  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: "Nucleic 
            Acids Res. 2016 Jan 4. pii: gkv1520."
COMMENT     Sequence Label: pQE30-hPolB
FEATURES             Location/Qualifiers
     source          1..4444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2..106)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     174..192
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(216..230,231..236,237..253,254..260)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    236..252
                     /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)."
     protein_bind    268..284
                     /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)."
     RBS             303..314
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             333..350
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1397..1491
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1535..2194
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1601..1620)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2259..2345
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2392..2414)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2500..2640
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2555..2574
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2655..2672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2826..3414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2906..2925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3585..4376,4377..4444,1))
                     /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     4208..4227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
       61 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
      121 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
      181 gtatcacgag gccctttcgt cttcacctcg agaaatcata aaaaatttat ttgctttgtg
      241 agcggataac aattataata gattcaattg tgagcggata acaatttcac acagaattca
      301 ttaaagagga gaaattaact atgagaggat cgcatcacca tcaccatcac ggatcctcta
      361 aacggaaggc gccgcaggag actctcaacg ggggaatcac cgacatgctc acagaactcg
      421 caaactttga gaagaacgtg agccaagcta tccacaagta caatgcttac agaaaagcag
      481 catctgttat agcaaaatac ccacacaaaa taaagagtgg agctgaagct aagaaattgc
      541 ctggagtagg aacaaaaatt gctgaaaaga ttgatgagtt tttagcaact ggaaaattac
      601 gtaaactgga aaagattcgg caggatgata cgagttcatc catcaatttc ctgactcgag
      661 ttagtggcat tggtccatct gctgcaagga agtttgtaga tgaaggaatt aaaacactag
      721 aagatctcag aaaaaatgaa gataaattga accatcatca gcgaattggg ctgaaatatt
      781 ttggggactt tgaaaaaaga attcctcgtg aagagatgtt acaaatgcaa gatattgtac
      841 taaatgaagt taaaaaagtg gattctgaat acattgctac agtctgtggc agtttcagaa
      901 gaggtgcaga gtccagtggt gacatggatg ttctcctgac ccatcccagc ttcacttcag
      961 aatcaaccaa acagccaaaa ctgttacatc aggttgtgga gcagttacaa aaggttcatt
     1021 ttatcacaga taccctgtca aagggtgaga caaagttcat gggtgtttgc cagcttccca
     1081 gtaaaaatga tgaaaaagaa tatccacaca gaagaattga tatcaggttg atacccaaag
     1141 atcagtatta ctgtggtgtt ctctatttca ctgggagtga tattttcaat aagaatatga
     1201 gggctcatgc cctagaaaag ggtttcacaa tcaatgagta caccatccgt cccttgggag
     1261 tcactggagt tgcaggagaa cccctgccag tggatagtga aaaagacatc tttgattaca
     1321 tccagtggaa ataccgggaa cccaaggacc tgagcgaata agtcgacctg cagccaagct
     1381 taattagctg agcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     1441 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagctag
     1501 cttggcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     1561 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     1621 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     1681 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     1741 tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     1801 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     1861 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     1921 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     1981 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     2041 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     2101 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     2161 acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc
     2221 ttaaacgcct ggggtaatga ctctctagct tgaggcatca aataaaacga aaggctcagt
     2281 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     2341 caaatccgcc ctctagagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2401 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2461 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     2521 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     2581 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     2641 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2701 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2761 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2821 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2881 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2941 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3001 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3061 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3121 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3181 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3241 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3301 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3361 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3721 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4441 ctca
//
