LOCUS       Exported                7043 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression of Taq polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    pOpen_taq
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7043)
  TITLE     COVID-19 Diagnostic Toolkit Enzymes
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        283..311
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    319..335
                     /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     324..346
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             364..2856
                     /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(2897..2914)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2897..2913)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2906..2928)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3144..3161)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3314..3400
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3492..3519
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3634..3653)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3753..3775
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3813..3831)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3887..3964
                     /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             3965..5047
                     /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(3984..4003)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        5160..5264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5265..6125
                     /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(5483..5502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6296..6884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6785..6804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7038..12
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
       61 ccgattcatt aatgcagaat taattctcat gtttgacagc ttatcatcga ctgcacggtg
      121 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
      181 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
      241 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgttgacaat taatcatcgg
      301 ctcgtataat gtgtggaatt gtgagcggat aacaatttca cacaggaaac agcgatgaat
      361 tcggggatgc tgcccctctt tgagcccaag ggccgggtcc tcctggtgga cggccaccac
      421 ctggcctacc gcaccttcca cgccctgaag ggcctcacca ccagccgggg ggagccggtg
      481 caggcggtct acggcttcgc caagagcctc ctcaaggccc tcaaggagga cggggacgcg
      541 gtgatcgtgg tctttgacgc caaggccccc tccttccgcc acgaggccta cggggggtac
      601 aaggcgggcc gggcccccac gccggaggac tttccccggc aactcgccct catcaaggag
      661 ctggtggacc tcctggggct ggcgcgcctc gaggtcccgg gctacgaggc ggacgacgtc
      721 ctggccagcc tggccaagaa ggcggaaaag gagggctacg aggtccgcat cctcaccgcc
      781 gacaaagacc tttaccagct cctttccgac cgcatccacg tcctccaccc cgaggggtac
      841 ctcatcaccc cggcctggct ttgggaaaag tacggcctga ggcccgacca gtgggccgac
      901 taccgggccc tgaccgggga cgagtccgac aaccttcccg gggtcaaggg catcggggag
      961 aagacggcga ggaagcttct ggaggagtgg gggagcctgg aagccctcct caagaacctg
     1021 gaccggctga agcccgccat ccgggagaag atcctggccc acatggacga tctgaagctc
     1081 tcctgggacc tggccaaggt gcgcaccgac ctgcccctgg aggtggactt cgccaaaagg
     1141 cgggagcccg accgggagag gcttagggcc tttctggaga ggcttgagtt tggcagcctc
     1201 ctccacgagt tcggccttct ggaaagcccc aaggccctgg aggaggcccc ctggcccccg
     1261 ccggaagggg ccttcgtggg ctttgtgctt tcccgcaagg agcccatgtg ggccgatctt
     1321 ctggccctgg ccgccgccag ggggggccgg gtccaccggg cccccgagcc ttataaagcc
     1381 ctcagggacc tgaaggaggc gcgggggctt ctcgccaaag acctgagcgt tctggccctg
     1441 agggaaggcc ttggcctccc gcccggcgac gaccccatgc tcctcgccta cctcctggac
     1501 ccttccaaca ccacccccga gggggtggcc cggcgctacg gcggggagtg gacggaggag
     1561 gcgggggagc gggccgccct ttccgagagg ctcttcgcca acctgtgggg gaggcttgag
     1621 ggggaggaga ggctcctttg gctttaccgg gaggtggaga ggcccctttc cgctgtcctg
     1681 gcccacatgg aggccacggg ggtgcgcctg gacgtggcct atctcagggc cttgtccctg
     1741 gaggtggccg aggagatcgc ccgcctcgag gccgaggtct tccgcctggc cggccacccc
     1801 ttcaacctca actcccggga ccagctggaa agggtcctct ttgacgagct agggcttccc
     1861 gccatcggca agacggagaa gaccggcaag cgctccacca gcgccgccgt cctggaggcc
     1921 ctccgcgagg cccaccccat cgtggagaag atcctgcagt accgggagct caccaagctg
     1981 aagagcacct acattgaccc cttgccggac ctcatccacc ccaggacggg ccgcctccac
     2041 acccgcttca accagacggc cacggccacg ggcaggctaa gtagctccga tcccaacctc
     2101 cagaacatcc ccgtccgcac cccgcttggg cagaggatcc gccgggcctt catcgccgag
     2161 gaggggtggc tattggtggc cctggactat agccagatag agctcagggt gctggcccac
     2221 ctctccggcg acgagaacct gatccgggtc ttccaggagg ggcgggacat ccacacggag
     2281 accgccagct ggatgttcgg cgtcccccgg gaggccgtgg accccctgat gcgccgggcg
     2341 gccaagacca tcaacttcgg ggtcctctac ggcatgtcgg cccaccgcct ctcccaggag
     2401 ctagccatcc cttacgagga ggcccaggcc ttcattgagc gctactttca gagcttcccc
     2461 aaggtgcggg cctggattga gaagaccctg gaggagggca ggaggcgggg gtacgtggag
     2521 accctcttcg gccgccgccg ctacgtgcca gacctagagg cccgggtgaa gagcgtgcgg
     2581 gaggcggccg agcgcatggc cttcaacatg cccgtccagg gcaccgccgc cgacctcatg
     2641 aagctggcta tggtgaagct cttccccagg ctggaggaaa tgggggccag gatgctcctt
     2701 caggtccacg acgagctggt cctcgaggcc ccaaaagaga gggcggaggc cgtggcccgg
     2761 ctggccaagg aggtcatgga gggggtgtat cccctggccg tgcccctgga ggtggaggtg
     2821 gggatagggg aggactggct ctccgccaag gagtgataga tcctctagag tcgacctgca
     2881 ggcatgcaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2941 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3001 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
     3061 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat tccctgtttt
     3121 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     3181 ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac
     3241 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     3301 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3361 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     3421 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     3481 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactcttcc
     3541 tgtcgtcata tctacaagcc atccccccac agatacggta aactagcctc gtttttgcat
     3601 caggaaagca gggaatttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3661 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3721 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3781 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3841 taatgtcatg ataataatgg tttcttagac gtgaggttct gtacccgaca ccatcgaatg
     3901 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     3961 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     4021 cgtttcccgc gtggtgaacc aggcaagcca cgtttctgcg aaaacgcggg aaaaagtgga
     4081 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     4141 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     4201 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     4261 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     4321 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     4381 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     4441 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     4501 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     4561 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     4621 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     4681 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     4741 gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag
     4801 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     4861 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     4921 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     4981 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     5041 gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccc caggctttac
     5101 actttatgct tccgacctgc aagaacctca cgtcaggtgg cacttttcgg ggaaatgtgc
     5161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6421 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6841 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6901 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6961 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7021 gccgaacgac cgagcgcagc gag
//
