LOCUS       sequence-198041-        6172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Taq DNA polymerase under WT T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6172)
  AUTHORS   Abil Z, Ellefson JW, Gollihar JD, Watkins E, Ellington AD
  TITLE     Compartmentalized partnered replication for the directed evolution 
            of genetic parts and circuits.
  JOURNAL   Nat Protoc. 2017 Dec;12(12):2493-2512. doi: 10.1038/nprot.2017.119. 
            Epub 2017 Nov 9.
  PUBMED    29120463
REFERENCE   2  (bases 1 to 6172)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nprot.2017.119"; journalName: "Nat Protoc"; date: 
            "2017-12"; volume: "12"; issue: "12"; pages: "2493-2512"
COMMENT     Sequence Label: pACYC-Taq
FEATURES             Location/Qualifiers
     source          1..6172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1444..1468
                     /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     complement(1468..1487)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1469..1487)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1533..1610
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1611..2693
                     /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(1630..1649)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      2918..3463
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     3364..3383
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3580..3597
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3989..4091
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4092..4751
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4158..4177)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(4972..5019)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(5030..6172,1..1350))
                     /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"
ORIGIN
        1 gtccaggcgc acccccgtgg cctccatgtg ggccaggaca gcggaaaggg gcctctccac
       61 ctcccggtaa agccaaagga gcctctcctc cccctcaagc ctcccccaca ggttggcgaa
      121 gagcctctcg gaaagggcgg cccgctcccc cgcctcctcc gtccactccc cgccgtagcg
      181 ccgggccacc ccctcggggg tggtgttgga agggtccagg aggtaggcga ggagcatggg
      241 gtcgtcgccg ggcgggaggc caaggccttc cctcagggcc agaacgctca ggtctttggc
      301 gagaagcccc cgcgcctcct tcaggtccct gagggcttta taaggctcgg gggcccggtg
      361 gacccggccc cccctggcgg cggccagggc cagaagatcg gcccacatgg gctccttgcg
      421 ggaaagcaca aagcccacga aggccccttc cggcgggggc cagggggcct cctccagggc
      481 cttggggctt tccagaaggc cgaactcgtg gaggaggctg ccaaactcaa gcctctccag
      541 aaaggcccta agcctctccc ggtcgggctc ccgccttttg gcgaagtcca cctccagggg
      601 caggtcggtg cgcaccttgg ccaggtccca ggagagcttc agatcgtcca tgtgggccag
      661 gatcttctcc cggatggcgg gcttcagccg gtccaggttc ttgaggaggg cttccaggct
      721 cccccactcc tccagaagct tcctcgccgt cttctccccg atgcccttga ccccgggaag
      781 gttgtcggac tcgtccccgg tcagggcccg gtagtcggcc cactggtcgg gcctcaggcc
      841 gtacttttcc caaagccagg ccggggtgat gaggtacccc tcggggtgga ggacgtggat
      901 gcggtcggaa aggagctggt aaaggtcttt gtcggcggtg aggatgcgga cctcgtagcc
      961 ctccttttcc gccttcttgg ccaggctggc caggacgtcg tccgcctcgt agcccgggac
     1021 ctcgaggcgc gccagcccca ggaggtccac cagctccttg atgagggcga gttgccgggg
     1081 aaagtcctcc ggcgtggggg cccggcccgc cttgtacccc ccgtaggcct cgtggcggaa
     1141 ggagggggcc ttggcgtcaa agaccacgat caccgcgtcc ccgtcctcct tgagggcctt
     1201 gaggaggctc ttggcgaagc cgtagaccgc ctgcaccggc tccccccggc tggtggtgag
     1261 gcccttcagg gcgtggaagg tgcggtaggc caggtggtgg ccgtccacca ggaggacccg
     1321 gcccttgggc tcaaagaggg gcagcatccc cgaattcata aagcttggcg taatcatggt
     1381 catccccatc tagtagtcct gtgtgactct agaattaaag ttaaacaaaa ttatttctac
     1441 aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt tcctaatgca
     1501 ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc aaaacctttc
     1561 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     1621 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     1681 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     1741 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     1801 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     1861 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     1921 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     1981 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     2041 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     2101 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     2161 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     2221 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     2281 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     2341 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     2401 ttggtgcgga catctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     2461 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     2521 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     2581 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2641 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2701 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga
     2761 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactaacat gagaattaca
     2821 acttatatcg tatggggctg acttcaggtg ctacatttga agagataaat tgcactgaaa
     2881 tctagaaata ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg
     2941 taatctcttg ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct
     3001 gagctaccaa ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg
     3061 tcctttcagt ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta
     3121 ccagtggctg ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag
     3181 ttaccggata aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg
     3241 gagcgaactg cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa
     3301 cagcggaatg acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc
     3361 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg
     3421 tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc
     3481 cctctcactt ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg
     3541 taagccattt ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa
     3601 gcggaatata tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg
     3661 ccacatgaag cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact
     3721 ccgctagcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt cagtagctga
     3781 acaggaggga cagctgatag aaacagaagc cactggagca cctcaaaaac accatcatac
     3841 actaaatcag taagttggca gcatcacccg acgcactttg cgccgaataa atacctgtga
     3901 cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc gggaagccct
     3961 gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc aactttcacc
     4021 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     4081 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     4141 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     4201 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     4261 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggagttc cgtatggcaa
     4321 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     4381 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     4441 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     4501 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     4561 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcactatg ggcaaatatt
     4621 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     4681 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     4741 gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt tgctacgcct
     4801 gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg acccggtcgt
     4861 cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta ccggtttatt
     4921 gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggtttca gcaaaaaacc
     4981 cctcaagacc cgtttagagg ccccaagggg ttatgctagt ctaattctat cactccttgg
     5041 cggagagcca gtcctcccct atccccacct ccacctccag gggcacggcc aggggataca
     5101 ccccctccat gacctccttg gccagccggg ccacggcctc cgccctctct tttggggcct
     5161 cgaggaccag ctcgtcgtgg acctgaagga gcatcctggc ccccatttcc tccagcctgg
     5221 ggaagagctt caccatagcc agcttcatga ggtcggcggc ggtgccctgg acgggcatgt
     5281 tgaaggccat gcgctcggcc gcctcccgca cgctcttcac ccgggcctct aggtctggca
     5341 cgtagcggcg gcggccgaag agggtctcca cgtacccccg cctcctgccc tcctccaggg
     5401 tcttctcaat ccaggcccgc accttgggga agctctgaaa gtagcgctca atgaaggcct
     5461 gggcctcctc gtaagggatg gctagctcct gggagaggcg gtgggccgac atgccgtaga
     5521 ggaccccgaa gttgatggtc ttggccgccc ggcgcatcag ggggtccacg gcctcccggg
     5581 ggacgccgaa catccagctg gcggtctccg tgtggatgtc ccgcccctcc tggaagaccc
     5641 ggatcaggtt ctcgtcgccg gagaggtggg ccagcaccct gagctctatc tggctatagt
     5701 ccagggccac caatagccac ccctcctcgg cgatgaaggc ccggcggatc ctctgcccaa
     5761 gcggggtgcg gacggggatg ttctggaggt tgggatcgga gctacttagc ctgcccgtgg
     5821 ccgtggccgt ctggttgaag cgggtgtgga ggcggcccgt cctggggtgg atgaggtccg
     5881 gcaaggggtc aatgtaggtg ctcttcagct tggtgagctc ccggtactgc aggatcttct
     5941 ccacgatggg gtgggcctcg cggagggcct ccaggacggc ggcgctggtg gagcgcttgc
     6001 cggtcttctc cgtcttgccg atggcgggaa gccctagctc gtcaaagagg accctttcca
     6061 gctggtcccg ggagttgagg ttgaaggggt ggccggccag gcggaagacc tcggcctcga
     6121 ggcgggcgat ctcctcggcc acctccaggg acaaggccct gagataggcc ac
//
