LOCUS       sequence-198045-        6161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Taq DNA polymerases with a premature stop codon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6161)
  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 6161)
  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.1Amb
FEATURES             Location/Qualifiers
     source          1..6161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1467..1491
                     /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(1491..1510)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1492..1510)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1556..1633
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1634..2716
                     /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(1653..1672)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      2941..3486
                     /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     3387..3406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3603..3620
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4012..4114
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4115..4774
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4181..4200)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(4995..5042)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(5053..6161,1..1384))
                     /codon_start=1
                     /product="thermostable DNA polymerase from Thermus 
                     aquaticus"
                     /label=Taq polymerase
                     /translation="GMLPLFEPKGRVLLVDGHHLAYRTFHALKGLTTSRGEPVQAVYGF
                     AKSLLKALKEDGDAVIVVFDAKAPSFRHEAYGGYKAGRAPTPEDFPRQLALIKELVDLL
                     GLARLEVPGYEADDVLASLAKKAEKEGYEVRILTADKDLYQLLSDRIHALHPEGYLITP
                     A*LWEKYGLRPDQWADYRALTGDESDNLPGVKGIGEKTARKLLEEWGSLEALLKNLDRL
                     KPAIREKILAHMDDLKLSWDLAKVRTDLPLEVDFAKRREPDRERLRAFLERLEFGSLLH
                     EFGLLESPKALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKAL
                     RDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEE
                     AGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALS
                     LEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAAVL
                     EALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSSSD
                     PNLQNIPVRTPLGQRIRRAFIAEEGWLLVALDYSQIELRVLAHLSGDENLIRVFQEGRD
                     IHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERY
                     FQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPVQG
                     TAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYPLA
                     VPLEVEVGIGEDWLSAKE"
ORIGIN
        1 ccacctccag ggacaaggcc ctgagatagg ccacgtccag gcgcaccccc gtggcctcca
       61 tgtgggccag gacagcggaa aggggcctct ccacctcccg gtaaagccaa aggagcctct
      121 cctccccctc aagcctcccc cacaggttgg cgaagagcct ctcggaaagg gcggcccgct
      181 cccccgcctc ctccgtccac tccccgccgt agcgccgggc caccccctcg ggggtggtgt
      241 tggaagggtc caggaggtag gcgaggagca tggggtcgtc gccgggcggg aggccaaggc
      301 cttccctcag ggccagaacg ctcaggtctt tggcgagaag cccccgcgcc tccttcaggt
      361 ccctgagggc tttataaggc tcgggggccc ggtggacccg gccccccctg gcggcggcca
      421 gggccagaag atcggcccac atgggctcct tgcgggaaag cacaaagccc acgaaggccc
      481 cttccggcgg gggccagggg gcctcctcca gggccttggg gctttccaga aggccgaact
      541 cgtggaggag gctgccaaac tcaagcctct ccagaaaggc cctaagcctc tcccggtcgg
      601 gctcccgcct tttggcgaag tccacctcca ggggcaggtc ggtgcgcacc ttggccaggt
      661 cccaggagag cttcagatcg tccatgtggg ccaggatctt ctcccggatg gcgggcttca
      721 gccggtccag gttcttgagg agggcttcca ggctccccca ctcctccaga agcttcctcg
      781 ccgtcttctc cccgatgccc ttgaccccgg gaaggttgtc ggactcgtcc ccggtcaggg
      841 cccggtagtc ggcccactgg tcgggcctca ggccgtactt ttcccaaagc taggccgggg
      901 tgatgaggta cccctcgggg tggagggcgt ggatgcggtc ggaaaggagc tggtaaaggt
      961 ctttgtcggc ggtgaggatg cggacctcgt agccctcctt ttccgccttc ttggccaggc
     1021 tggccaggac gtcgtccgcc tcgtagcccg ggacctcgag gcgcgccagc cccaggaggt
     1081 ccaccagctc cttgatgagg gcgagttgcc ggggaaagtc ctccggcgtg ggggcccggc
     1141 ccgccttgta ccccccgtag gcctcgtggc ggaaggaggg ggccttggcg tcaaagacca
     1201 cgatcaccgc gtccccgtcc tccttgaggg ccttgaggag gctcttggcg aagccgtaga
     1261 ccgcctgcac cggctccccc cggctggtgg tgaggccctt cagggcgtgg aaggtgcggt
     1321 aggccaggtg gtggccgtcc accaggagga cccggccctt gggctcaaag aggggcagca
     1381 tccccgaatt cataaagctt ggcgtaatca tggtcatccc catggtatat ctccttatta
     1441 aagttaaaca aaattatttc tacaggggaa ttgttatccg ctcacaattc ccctatagtg
     1501 agtcgtatta atttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac
     1561 catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     1621 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     1681 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     1741 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     1801 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     1861 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     1921 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     1981 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     2041 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     2101 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     2161 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     2221 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     2281 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     2341 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     2401 cattaccgag tccggtctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac
     2461 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     2521 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     2581 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     2641 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     2701 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg
     2761 gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     2821 gcatgactaa catgagaatt acaacttata tcgtatgggg ctgacttcag gtgctacatt
     2881 tgaagagata aattgcactg aaatctagaa atattttatc tgattaataa gatgatcttc
     2941 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
     3001 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
     3061 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
     3121 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
     3181 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
     3241 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
     3301 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
     3361 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
     3421 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
     3481 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
     3541 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
     3601 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
     3661 ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc
     3721 aacatagtaa gccagtatac actccgctag cgctgatgtc cggcggtgct tttgccgtta
     3781 cgcaccaccc cgtcagtagc tgaacaggag ggacagctga tagaaacaga agccactgga
     3841 gcacctcaaa aacaccatca tacactaaat cagtaagttg gcagcatcac ccgacgcact
     3901 ttgcgccgaa taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt
     3961 ccctgttgat accgggaagc cctgggccaa cttttggcga aaatgagacg ttgatcggca
     4021 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
     4081 gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     4141 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     4201 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     4261 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     4321 tcatccggag ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     4381 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     4441 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     4501 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     4561 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     4621 cgttttcact atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     4681 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     4741 acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc
     4801 ttaaacgcct ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc
     4861 gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc cgcttatgtc
     4921 tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg cttctcaaat
     4981 gcctgaggtt tcagcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     5041 agtctaattc tatcactcct tggcggagag ccagtcctcc cctatcccca cctccacctc
     5101 caggggcacg gccaggggat acaccccctc catgacctcc ttggccagcc gggccacggc
     5161 ctccgccctc tcttttgggg cctcgaggac cagctcgtcg tggacctgaa ggagcatcct
     5221 ggcccccatt tcctccagcc tggggaagag cttcaccata gccagcttca tgaggtcggc
     5281 ggcggtgccc tggacgggca tgttgaaggc catgcgctcg gccgcctccc gcacgctctt
     5341 cacccgggcc tctaggtctg gcacgtagcg gcggcggccg aagagggtct ccacgtaccc
     5401 ccgcctcctg ccctcctcca gggtcttctc aatccaggcc cgcaccttgg ggaagctctg
     5461 aaagtagcgc tcaatgaagg cctgggcctc ctcgtaaggg atggctagct cctgggagag
     5521 gcggtgggcc gacatgccgt agaggacccc gaagttgatg gtcttggccg cccggcgcat
     5581 cagggggtcc acggcctccc gggggacgcc gaacatccag ctggcggtct ccgtgtggat
     5641 gtcccgcccc tcctggaaga cccggatcag gttctcgtcg ccggagaggt gggccagcac
     5701 cctgagctct atctggctat agtccagggc caccaatagc cacccctcct cggcgatgaa
     5761 ggcccggcgg atcctctgcc caagcggggt gcggacgggg atgttctgga ggttgggatc
     5821 ggagctactt agcctgcccg tggccgtggc cgtctggttg aagcgggtgt ggaggcggcc
     5881 cgtcctgggg tggatgaggt ccggcaaggg gtcaatgtag gtgctcttca gcttggtgag
     5941 ctcccggtac tgcaggatct tctccacgat ggggtgggcc tcgcggaggg cctccaggac
     6001 ggcggcgctg gtggagcgct tgccggtctt ctccgtcttg ccgatggcgg gaagccctag
     6061 ctcgtcaaag aggacccttt ccagctggtc ccgggagttg aggttgaagg ggtggccggc
     6121 caggcggaag acctcggcct cgaggcgggc gatctcctcg g
//
