LOCUS       sequence-315991-        7043 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression of Taq polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Anal 
            Biochem. 1990 Dec . 191(2):396-400."
COMMENT     Sequence Label: pAKTaq
FEATURES             Location/Qualifiers
     source          1..7043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(131..136,137..152,153..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             join(5113..5181,5182..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
//
