LOCUS       Exported                7043 bp ds-DNA     circular SYN 01-MAY-2020
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 Monday, May 11, 2020 from SnapGene Viewer 5.1.0
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Anal 
            Biochem. 1990 Dec . 191(2):396-400."
FEATURES             Location/Qualifiers
     source          1..7043
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1031..1059
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1067..1083
                     /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     1072..1094
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1112..3604
                     /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(3645..3662)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3645..3661)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3654..3676)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3892..3909)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4062..4148
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4240..4267
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4382..4401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4501..4523
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4561..4579)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4635..4712
                     /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             4713..5795
                     /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(4732..4751)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        5908..6012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6013..6873
                     /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(6231..6250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagaatta attctcatgt
      841 ttgacagctt atcatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg
      901 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc
      961 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
     1021 aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa
     1081 caatttcaca caggaaacag cgatgaattc ggggatgctg cccctctttg agcccaaggg
     1141 ccgggtcctc ctggtggacg gccaccacct ggcctaccgc accttccacg ccctgaaggg
     1201 cctcaccacc agccgggggg agccggtgca ggcggtctac ggcttcgcca agagcctcct
     1261 caaggccctc aaggaggacg gggacgcggt gatcgtggtc tttgacgcca aggccccctc
     1321 cttccgccac gaggcctacg gggggtacaa ggcgggccgg gcccccacgc cggaggactt
     1381 tccccggcaa ctcgccctca tcaaggagct ggtggacctc ctggggctgg cgcgcctcga
     1441 ggtcccgggc tacgaggcgg acgacgtcct ggccagcctg gccaagaagg cggaaaagga
     1501 gggctacgag gtccgcatcc tcaccgccga caaagacctt taccagctcc tttccgaccg
     1561 catccacgtc ctccaccccg aggggtacct catcaccccg gcctggcttt gggaaaagta
     1621 cggcctgagg cccgaccagt gggccgacta ccgggccctg accggggacg agtccgacaa
     1681 ccttcccggg gtcaagggca tcggggagaa gacggcgagg aagcttctgg aggagtgggg
     1741 gagcctggaa gccctcctca agaacctgga ccggctgaag cccgccatcc gggagaagat
     1801 cctggcccac atggacgatc tgaagctctc ctgggacctg gccaaggtgc gcaccgacct
     1861 gcccctggag gtggacttcg ccaaaaggcg ggagcccgac cgggagaggc ttagggcctt
     1921 tctggagagg cttgagtttg gcagcctcct ccacgagttc ggccttctgg aaagccccaa
     1981 ggccctggag gaggccccct ggcccccgcc ggaaggggcc ttcgtgggct ttgtgctttc
     2041 ccgcaaggag cccatgtggg ccgatcttct ggccctggcc gccgccaggg ggggccgggt
     2101 ccaccgggcc cccgagcctt ataaagccct cagggacctg aaggaggcgc gggggcttct
     2161 cgccaaagac ctgagcgttc tggccctgag ggaaggcctt ggcctcccgc ccggcgacga
     2221 ccccatgctc ctcgcctacc tcctggaccc ttccaacacc acccccgagg gggtggcccg
     2281 gcgctacggc ggggagtgga cggaggaggc gggggagcgg gccgcccttt ccgagaggct
     2341 cttcgccaac ctgtggggga ggcttgaggg ggaggagagg ctcctttggc tttaccggga
     2401 ggtggagagg cccctttccg ctgtcctggc ccacatggag gccacggggg tgcgcctgga
     2461 cgtggcctat ctcagggcct tgtccctgga ggtggccgag gagatcgccc gcctcgaggc
     2521 cgaggtcttc cgcctggccg gccacccctt caacctcaac tcccgggacc agctggaaag
     2581 ggtcctcttt gacgagctag ggcttcccgc catcggcaag acggagaaga ccggcaagcg
     2641 ctccaccagc gccgccgtcc tggaggccct ccgcgaggcc caccccatcg tggagaagat
     2701 cctgcagtac cgggagctca ccaagctgaa gagcacctac attgacccct tgccggacct
     2761 catccacccc aggacgggcc gcctccacac ccgcttcaac cagacggcca cggccacggg
     2821 caggctaagt agctccgatc ccaacctcca gaacatcccc gtccgcaccc cgcttgggca
     2881 gaggatccgc cgggccttca tcgccgagga ggggtggcta ttggtggccc tggactatag
     2941 ccagatagag ctcagggtgc tggcccacct ctccggcgac gagaacctga tccgggtctt
     3001 ccaggagggg cgggacatcc acacggagac cgccagctgg atgttcggcg tcccccggga
     3061 ggccgtggac cccctgatgc gccgggcggc caagaccatc aacttcgggg tcctctacgg
     3121 catgtcggcc caccgcctct cccaggagct agccatccct tacgaggagg cccaggcctt
     3181 cattgagcgc tactttcaga gcttccccaa ggtgcgggcc tggattgaga agaccctgga
     3241 ggagggcagg aggcgggggt acgtggagac cctcttcggc cgccgccgct acgtgccaga
     3301 cctagaggcc cgggtgaaga gcgtgcggga ggcggccgag cgcatggcct tcaacatgcc
     3361 cgtccagggc accgccgccg acctcatgaa gctggctatg gtgaagctct tccccaggct
     3421 ggaggaaatg ggggccagga tgctccttca ggtccacgac gagctggtcc tcgaggcccc
     3481 aaaagagagg gcggaggccg tggcccggct ggccaaggag gtcatggagg gggtgtatcc
     3541 cctggccgtg cccctggagg tggaggtggg gataggggag gactggctct ccgccaagga
     3601 gtgatagatc ctctagagtc gacctgcagg catgcaagct tggcactggc cgtcgtttta
     3661 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3721 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3781 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3841 atttcacacc gcataaattc cctgttttgg cggatgagag aagattttca gcctgataca
     3901 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
     3961 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     4021 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     4081 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     4141 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     4201 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
     4261 gcctttttgc gtttctacaa actcttcctg tcgtcatatc tacaagccat ccccccacag
     4321 atacggtaaa ctagcctcgt ttttgcatca ggaaagcagg gaatttatgg tgcactctca
     4381 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4441 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4501 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4561 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4621 gaggttctgt acccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
     4681 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     4741 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     4801 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     4861 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     4921 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     4981 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     5041 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     5101 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     5161 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     5221 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     5281 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     5341 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     5401 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     5461 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     5521 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     5581 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     5641 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     5701 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5761 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     5821 ctcactcatt aggcacccca ggctttacac tttatgcttc cgacctgcaa gaacctcacg
     5881 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5941 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6001 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6061 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6121 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6181 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6241 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6301 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6361 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6421 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6481 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6541 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6601 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6661 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6721 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6781 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6841 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6901 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6961 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7021 gtagaaaaga tcaaaggatc ttc
//
