LOCUS       sequence-471802-        5661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses native TaTasR TIGR locus, TasR has premature stop codon. 
            CmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5661)
  AUTHORS   Faure G, Saito M, Wilkinson ME, Quinones-Olvera N, Xu P, 
            Flam-Shepherd D, Kim S, Reddy N, Zhu S, Evgeniou L, Koonin EV, 
            Macrae RK, Zhang F
  TITLE     TIGR-Tas: A family of modular RNA-guided DNA-targeting systems in 
            prokaryotes and their viruses.
  JOURNAL   Science. 2025 Feb 27:eadv9789. doi: 10.1126/science.adv9789.
  PUBMED    40014690
REFERENCE   2  (bases 1 to 5661)
  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"; journalName: "Science. 
            2025 Feb 27:eadv9789. doi: 10.1126/science.adv9789."
COMMENT     Sequence Label: pMIIS794
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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(2099..2117)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2113..2160
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2381..3040)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2955..2974
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3041..3143)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3535..3552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3669..4214)
                     /direction=LEFT
                     /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     complement(3749..3768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4439..5521)
                     /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     5483..5502
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5522..5599)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5645..5661,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5645..5661,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatgtgat taagccatac ccccactgta acccactgtc
      121 cccgtgcgag ccaaaatcaa cgtgaaaccc atcatcaacg tgcaaataaa cctcattaat
      181 caatgaggtt tattttttgt attgacatat ttctttattg aaaatacaat tttgaaatca
      241 aacttctgaa aaggcaataa attacatgga taacgatatc acaatattgg cggtcgattg
      301 gtctcatgaa gaacgtaaac ttgcaatttt tgatggcaag aagataagga aaaaacttcc
      361 tgagccttct agcgatgtaa taattgtggc tgaaaatatt cctcaaaaat atgccgctcc
      421 gttcatcgag gtaggggcaa aagttcttag atgttcgaca aacgccacag cagatgcaag
      481 aaaaaactat caaaagaaag tcgatgctgc ttttgcgaaa acgacgaaaa tgattctaaa
      541 gtgatttggg cgttatatca aacacaccca gaattgtttc gtgaaatgaa gcttgagcca
      601 cccctttctt cttattatgc tatttttaaa gattaccaag aggtaagaat tagaactggc
      661 aatcgactct attcagatcg cactgatgca atggaagagt tttttaaaat agttaagaaa
      721 ggtgaacatg aactaaaaaa ggctgtcgat aaagaattag aaaatcatcc ggtctatacg
      781 cagtggttac agcatattaa aggtattggc ccggtcgttg ccgggggatt gatatcgtta
      841 attggagata tagatcgctt cgacagcgtt tctaaattat gggcctatgc aggttatagt
      901 gttgataatg gaaaagtgca aaagcgtaaa aaaggtgtag cttcaaattg gaaaaataaa
      961 attcgcacac attgttacaa cattgtagat tcatttatca aacaacgcac ttcagtttat
     1021 cgtgagttgt atgatgccga aaaagctcgt caacgtccaa aggtagaatc cgatggtcat
     1081 gctcacaacc gtgctgtaag aaaagttgct aaggtatttt tacaacatta ttgggtagtt
     1141 tctagagaat tagcagggtt ttctgtgtct aaaccttgga ttttagaaca tggtgggcac
     1201 gttgattaca tcaaaccacc acattggaat aaagtagaaa ttaagccatg caagccctga
     1261 aacccattgc cttagtgcga gccatcatat atatggaacc catgccgcag atacgagcca
     1321 agcgaaaaat gaaacccaac gtcgctttgc gagccaaact acatatgaaa cccacaatgg
     1381 agatgcgagc caaactacat atgaaaccca tgctgctaga tgcgagctaa gcggaaaatg
     1441 aaacccaaca ccgatttgcg agccaaacga tctatgaaac ccaaggtcta gttgcgagcc
     1501 agaagccctt tgaaacccat catcaacgtg cgaaaaagcc cgtttttcaa cgggctttta
     1561 tcttttcata acaagttctc ttactctatc gtaaacacca caatattgac atttaagtat
     1621 taaattagtg tctgtggcat tagaatactt atcttgagta aaactaatga ctttatacac
     1681 atgatgccca gatagacaac aatataaata ttttataagg ctcataacat atttatgaaa
     1741 aacgggcttt tttattatac ataacactat gttatacttc aaaaagtggt tgttagaatt
     1801 tatggggtct acaggagttg tgtttgatcc caaacaaaaa tcaagagact ggaattggct
     1861 cggtgccccc ggctccactg gtgtctcccc taaagaagac ccaattggtt gtaaaaagaa
     1921 aaagaagaaa aagaagaaat gagttggttt aacgcactat ctaacccttt tcaacaaagt
     1981 gttaaaaaat ttatctacga acttatcgga ccagaacgat atatcaacca tgatcccatt
     2041 gtaggtctgc tggctaaaaa tgttagcact gccgaagagt atgactaggc tgctgccacc
     2101 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     2161 ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg
     2221 gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg
     2281 tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     2341 aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact
     2401 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     2461 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     2521 ccatagtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg
     2581 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     2641 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     2701 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     2761 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     2821 ggaactccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     2881 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     2941 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     3001 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     3061 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     3121 tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc
     3181 cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta
     3241 tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt actgatttag tgtatgatgg
     3301 tgtttttgag gtgctccagt ggcttctgtt tctatcagct gtccctcctg ttcagctact
     3361 gacggggtgg tgcgtaacgg caaaagcacc gccggacatc agcgctagcg gagtgtatac
     3421 tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa
     3481 aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct
     3541 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg
     3601 cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca
     3661 aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat
     3721 cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc
     3781 ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg
     3841 cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga
     3901 ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct
     3961 tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt
     4021 tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg
     4081 gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct
     4141 tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac
     4201 caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag atttcagtgc
     4261 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct
     4321 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4381 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4501 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4561 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4621 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4681 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     4741 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     4801 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     4861 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     4921 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     4981 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5041 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5101 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5161 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5221 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5281 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5341 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5401 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5461 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5521 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5581 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     5641 aaattaatac gactcactat a
//
