LOCUS Exported 7043 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Bacterial expression of Taq polymerase. ACCESSION . VERSION . KEYWORDS pAKTaq SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7043) AUTHORS Engelke DR, Krikos A, Bruck ME, Ginsburg D TITLE Purification of Thermus aquaticus DNA polymerase expressed in Escherichia coli. JOURNAL Anal Biochem. 1990 Dec . 191(2):396-400. PUBMED 2085185 REFERENCE 2 (bases 1 to 7043) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7043 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 131..159 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 167..183 /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)." primer_bind 172..194 /label=M13/pUC Reverse /note="In lacZ gene" CDS 212..2704 /codon_start=1 /product="thermostable DNA polymerase from Thermus aquaticus" /label=Taq polymerase /translation="GMLPLFEPKGRVLLVDGHHLAYRTFHALKGLTTSRGEPVQAVYGF AKSLLKALKEDGDAVIVVFDAKAPSFRHEAYGGYKAGRAPTPEDFPRQLALIKELVDLL GLARLEVPGYEADDVLASLAKKAEKEGYEVRILTADKDLYQLLSDRIHVLHPEGYLITP AWLWEKYGLRPDQWADYRALTGDESDNLPGVKGIGEKTARKLLEEWGSLEALLKNLDRL KPAIREKILAHMDDLKLSWDLAKVRTDLPLEVDFAKRREPDRERLRAFLERLEFGSLLH EFGLLESPKALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKAL RDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEE AGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALS LEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAAVL EALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSSSD PNLQNIPVRTPLGQRIRRAFIAEEGWLLVALDYSQIELRVLAHLSGDENLIRVFQEGRD IHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERY FQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPVQG TAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYPLA VPLEVEVGIGEDWLSAKE" primer_bind complement(2745..2762) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2745..2761) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2754..2776) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(2992..3009) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 3162..3248 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 3340..3367 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind complement(3482..3501) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3601..3623 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3661..3679) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3735..3812 /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." CDS 3813..4895 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(3832..3851) /label=LacI-R /note="LacI, reverse primer" promoter 5008..5112 /gene="bla" /label=AmpR promoter CDS 5113..5973 /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 complement(5331..5350) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6144..6732 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6633..6652 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6886..6903 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc 61 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg 121 aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa 181 caatttcaca caggaaacag cgatgaattc ggggatgctg cccctctttg agcccaaggg 241 ccgggtcctc ctggtggacg gccaccacct ggcctaccgc accttccacg ccctgaaggg 301 cctcaccacc agccgggggg agccggtgca ggcggtctac ggcttcgcca agagcctcct 361 caaggccctc aaggaggacg gggacgcggt gatcgtggtc tttgacgcca aggccccctc 421 cttccgccac gaggcctacg gggggtacaa ggcgggccgg gcccccacgc cggaggactt 481 tccccggcaa ctcgccctca tcaaggagct ggtggacctc ctggggctgg cgcgcctcga 541 ggtcccgggc tacgaggcgg acgacgtcct ggccagcctg gccaagaagg cggaaaagga 601 gggctacgag gtccgcatcc tcaccgccga caaagacctt taccagctcc tttccgaccg 661 catccacgtc ctccaccccg aggggtacct catcaccccg gcctggcttt gggaaaagta 721 cggcctgagg cccgaccagt gggccgacta ccgggccctg accggggacg agtccgacaa 781 ccttcccggg gtcaagggca tcggggagaa gacggcgagg aagcttctgg aggagtgggg 841 gagcctggaa gccctcctca agaacctgga ccggctgaag cccgccatcc gggagaagat 901 cctggcccac atggacgatc tgaagctctc ctgggacctg gccaaggtgc gcaccgacct 961 gcccctggag gtggacttcg ccaaaaggcg ggagcccgac cgggagaggc ttagggcctt 1021 tctggagagg cttgagtttg gcagcctcct ccacgagttc ggccttctgg aaagccccaa 1081 ggccctggag gaggccccct ggcccccgcc ggaaggggcc ttcgtgggct ttgtgctttc 1141 ccgcaaggag cccatgtggg ccgatcttct ggccctggcc gccgccaggg ggggccgggt 1201 ccaccgggcc cccgagcctt ataaagccct cagggacctg aaggaggcgc gggggcttct 1261 cgccaaagac ctgagcgttc tggccctgag ggaaggcctt ggcctcccgc ccggcgacga 1321 ccccatgctc ctcgcctacc tcctggaccc ttccaacacc acccccgagg gggtggcccg 1381 gcgctacggc ggggagtgga cggaggaggc gggggagcgg gccgcccttt ccgagaggct 1441 cttcgccaac ctgtggggga ggcttgaggg ggaggagagg ctcctttggc tttaccggga 1501 ggtggagagg cccctttccg ctgtcctggc ccacatggag gccacggggg tgcgcctgga 1561 cgtggcctat ctcagggcct tgtccctgga ggtggccgag gagatcgccc gcctcgaggc 1621 cgaggtcttc cgcctggccg gccacccctt caacctcaac tcccgggacc agctggaaag 1681 ggtcctcttt gacgagctag ggcttcccgc catcggcaag acggagaaga ccggcaagcg 1741 ctccaccagc gccgccgtcc tggaggccct ccgcgaggcc caccccatcg tggagaagat 1801 cctgcagtac cgggagctca ccaagctgaa gagcacctac attgacccct tgccggacct 1861 catccacccc aggacgggcc gcctccacac ccgcttcaac cagacggcca cggccacggg 1921 caggctaagt agctccgatc ccaacctcca gaacatcccc gtccgcaccc cgcttgggca 1981 gaggatccgc cgggccttca tcgccgagga ggggtggcta ttggtggccc tggactatag 2041 ccagatagag ctcagggtgc tggcccacct ctccggcgac gagaacctga tccgggtctt 2101 ccaggagggg cgggacatcc acacggagac cgccagctgg atgttcggcg tcccccggga 2161 ggccgtggac cccctgatgc gccgggcggc caagaccatc aacttcgggg tcctctacgg 2221 catgtcggcc caccgcctct cccaggagct agccatccct tacgaggagg cccaggcctt 2281 cattgagcgc tactttcaga gcttccccaa ggtgcgggcc tggattgaga agaccctgga 2341 ggagggcagg aggcgggggt acgtggagac cctcttcggc cgccgccgct acgtgccaga 2401 cctagaggcc cgggtgaaga gcgtgcggga ggcggccgag cgcatggcct tcaacatgcc 2461 cgtccagggc accgccgccg acctcatgaa gctggctatg gtgaagctct tccccaggct 2521 ggaggaaatg ggggccagga tgctccttca ggtccacgac gagctggtcc tcgaggcccc 2581 aaaagagagg gcggaggccg tggcccggct ggccaaggag gtcatggagg gggtgtatcc 2641 cctggccgtg cccctggagg tggaggtggg gataggggag gactggctct ccgccaagga 2701 gtgatagatc ctctagagtc gacctgcagg catgcaagct tggcactggc cgtcgtttta 2761 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc 2821 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg 2881 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt 2941 atttcacacc gcataaattc cctgttttgg cggatgagag aagattttca gcctgataca 3001 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc 3061 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag 3121 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc 3181 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta 3241 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg 3301 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg 3361 gcctttttgc gtttctacaa actcttcctg tcgtcatatc tacaagccat ccccccacag 3421 atacggtaaa ctagcctcgt ttttgcatca ggaaagcagg gaatttatgg tgcactctca 3481 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg 3541 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct 3601 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg 3661 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt 3721 gaggttctgt acccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc 3781 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg 3841 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg 3901 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca 3961 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca 4021 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac 4081 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg 4141 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg 4201 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg 4261 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg 4321 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa 4381 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa 4441 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca 4501 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg 4561 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg 4621 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca 4681 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg 4741 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc 4801 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 4861 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag 4921 ctcactcatt aggcacccca ggctttacac tttatgcttc cgacctgcaa gaacctcacg 4981 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata 5041 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga 5101 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca 5161 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat 5221 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag 5281 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc 5341 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct 5401 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca 5461 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt 5521 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat 5581 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt 5641 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta 5701 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga 5761 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt 5821 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc 5881 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct 5941 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata 6001 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt 6061 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 6121 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 6181 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 6241 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg 6301 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 6361 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 6421 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 6481 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 6541 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 6601 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 6661 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 6721 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 6781 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 6841 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc 6901 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat 6961 taatgcagaa ttaattctca tgtttgacag cttatcatcg actgcacggt gcaccaatgc 7021 ttctggcgtc aggcagccat cgg //