LOCUS       sequence-471794-        6023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses native FpTasA TIGR locus. CmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6023)
  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 6023)
  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: pMIIS644
FEATURES             Location/Qualifiers
     source          1..6023
                     /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)."
     CDS             complement(2054..2065)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2461..2479)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2475..2522
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2743..3402)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3317..3336
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3403..3505)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3897..3914)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4031..4576)
                     /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(4111..4130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4801..5883)
                     /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     5845..5864
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5884..5961)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(6007..6023,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(6007..6023,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatgcaac ccatggtaag gcccacgata gcatgtggga
      121 tgttgcaaga gatctgtgta ccctaaggtc tctaggcatt gatgtggggg acacaattat
      181 aatcccagaa atgagtgaag acgattactc tgctagaaat ctggaggcca gagagctcgg
      241 atggctacac cagataggtg agcagggttt gacgttctag gttggaggag ttgcaatgtt
      301 tagatgcata gcaaaatgtc ctgaactgga tgctgatggg gttacacatt gcacgtgcaa
      361 tgatcccgag gagatgtacg agcgttgggt cgatggttgt cccgtcggga atactcctaa
      421 gttcgttgag gtggacgcgg ctaagtaaac aggggtttat gctcatatat acatgaaagg
      481 gtaaagccca atccaatata gacagaaaga aggatttgtt ttatggacat tcaaaagaac
      541 aggattcgca acatcgttgg tggcatctac gacatccaga aactccgtat cgctactggc
      601 aatcgcatcg tagccagcct gcggccaggc ctggtggagg aggtcaagga aggtgaggag
      661 gacaccaagt accttcctgc tattctttct gagtacagac gcatcaccga ttacttcgtt
      721 tccgagtttg agggtcgtgg aagcattgaa aaggccatca cccctaacaa tccggagtac
      781 atcaagagcc gccttgacta tgatctggtg acatcctaca agcggctgct ggagacggag
      841 gaggggctca cgaaggttgc tgagcgtgag gtcaaagcac atccaatgtg ggatgccttc
      901 tttgcagggg taaaaggctg cggaccgctc atgtccgccg tgtgcctcgc gtacttcgat
      961 ccctacaagg cgcgccacgc ttcctccttc tggcgctatg ccggcctgga cgtgcagcgc
     1021 gaccctgaca aggataagat gcgtggcgtt ggcaagtggt acactgagga gcgcccttac
     1081 atcgataagg acgggaagga gcagatgaag aagtcgctca cctacaaccc gttcctcaaa
     1141 accaagctgg ttggtgtcct cggctctgca ttcctgcggg ccaaagactc ctactatggt
     1201 aaggtttact acgattacaa aaatcgcctg gataaccggg atgaggactt ctcccctatt
     1261 gttaagcacc gcatggccac ccgttacgcc gtgaagatgt tcctgcggga tatgtgggtc
     1321 gtgtggcgtg agctggaggg ccttgaggtc accgagcctt atgaggtcgc taagttgggt
     1381 cacaagcccc accactcatc ttaaccgtgg cagcgtagta acccctatat gcaaagtgtg
     1441 ccttatctat taagtagtcc attagacagt agtgtacctt atctcatgag aaaccctgtc
     1501 gcactcagtg taccagtctg aacaagtaac ccaagctgct ttagtgtacc gtatggcaca
     1561 agcaatccac gatattcaag tgtaccttac ggcatgagta acccagcctc ctcaagtgta
     1621 ccgccgaaaa aaagaagccc attgtgaggg agtgtaccag aaagctctag taaccctaat
     1681 gcagcgagtg taccagaaag cccctagtac ccctaaatca gcaagtgtgc ccaattagca
     1741 aagaagtcct atatgacgca gtgtacctaa tccgctaagt aatccgctcg accttagtgt
     1801 atctttcggc atcagtaatc cttgacaaga aagtgtacct tagcacttga gtaaccctcg
     1861 aatcaggagt gtgccacttt cacggagtag cccaaaaatc gggagcgtgc ctgataagcc
     1921 aagaagccca tttgggagta gtgtaccttt actggttaga agccctgttg cctcaagtgt
     1981 acccgcgacc gcaagcaatc cagtacttgt tagtgtaccg tgttggagta gcaaccctta
     2041 caagtaaagt gtaccttcct tcgatagtaa cccagtaaca ctgagtgtgc cctattgtat
     2101 caggaatcct ttgcgcatga gtgtaccgtt cgatgaaaga acccctacgg cgaggagtgt
     2161 gccagtttga gctagtaacc caaaactaag aagtgtacct aacgatccaa gaagccctaa
     2221 gatgtgtagt gtgcccaaca ggtcaagaaa ccctatgaga gctagtgtgc ccatataata
     2281 cagaagagag tgttagagag gaagaacaat gtttaaatta aaaactgtgg gagcggtcac
     2341 gctggcatcg ctgctactcc tttcggggtg ttcaagtaac agcccagagg atgagataga
     2401 accagtaaac aacacgctac cctatgactc acgcacagtt accacactag gctgctgcca
     2461 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2521 tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac
     2581 cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg
     2641 ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta
     2701 gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca
     2761 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     2821 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     2881 gcccatagtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     2941 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     3001 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     3061 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     3121 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     3181 acggaactcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     3241 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     3301 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     3361 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     3421 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     3481 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt
     3541 cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg
     3601 tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat
     3661 ggtgtttttg aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta
     3721 ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat
     3781 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     3841 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     3901 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     3961 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     4021 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     4081 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     4141 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     4201 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     4261 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     4321 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     4381 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     4441 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     4501 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     4561 accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt
     4621 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt
     4681 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     4741 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     4801 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4861 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     4921 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     4981 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     5041 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     5101 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     5161 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     5221 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     5281 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     5341 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     5401 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     5461 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     5521 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     5581 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     5641 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     5701 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     5761 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     5821 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     5881 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     5941 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     6001 ggaaattaat acgactcact ata
//
