LOCUS Exported 4444 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION bacterial expression of human DNA Polymerase beta (POLB). ACCESSION . VERSION . KEYWORDS pQE30-hPolB 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4444 /organism="synthetic DNA construct" /mol_type="other DNA" 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 216..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(3585..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 //