LOCUS       sequence-472909-        5579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses native SpTasH TIGR locus. CmR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5579)
  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 5579)
  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: pMIIS638
FEATURES             Location/Qualifiers
     source          1..5579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1649..1673
                     /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(1673..1692)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1674..1692)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1738..1815
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1816..2898
                     /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(1835..1854)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3123..3668
                     /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     3569..3588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3785..3802
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4194..4296
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4297..4956
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4363..4382)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(5177..5224)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5220..5238
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 atatggctag cttacctttg atgcctgcct tgtccgtaat ggctcgctta atccacctgg
       61 ttgtctatag gctgatggct cgcttcccgc ttatggttgt cttcaggata gtgactcgct
      121 tttcgcgctt tgggtgtttt tccaatcatg gctcgcttgt tccggttggt tgtcttaggc
      181 cgtgtggctc gctttcaatc gatgattgtt ttctccgctt taactcgctc gcggcacttg
      241 gttgtcttct cttgcttgac tcgcttcacg cggttggttg tcttgtgtga attggctcgc
      301 ttcgtctccg tggttgtctt taacggaatg actaaggcga cccgaaggcc gcccgttatt
      361 tcaccattcc caaccgcgct cttgcggctg gacaatgcca gtgtgcccca gtttttcctg
      421 cacgtacagc gggcgcgtat ccagtccggc aagctcacgg cctacgaacc agtaatctgc
      481 cagaaagtgc ttcatcactg cccggagcgc tgccccgtgc cggtggctcg gcttggtgtc
      541 cttccacgcg cattcgatca actgcccttg cgtgttgcgg gacttagtga ccttttcgga
      601 taccgcaagc cgctccttgg tctgctcata cacagtgcgg taagggcact tgcgattctt
      661 aatcatgctg tttgccatgt tccataccat ggtgcgcagc gtcttgtttc ctccgcccgc
      721 ctcgcctttg gtgtaacggt cgtgtgaagg cttgtcgatc ccgatgtatg cccacagcgc
      781 agaggctgac ttggcctttt cgagatcaac gtacgtttgc aaacccgcca cagtgatagg
      841 gccaacacca ggcacgccca tggccgcctg agcaagcggg tcttttgcgt gcttgatgtg
      901 cttggtcact tcccggtcaa tctgagcaag gcgcttattg gatgcgtcca gcgtgtcttg
      961 caggaattgc tcggtgctct ggcgtgcgtg atcggtctgc cgctggtacg caagaagctg
     1021 gttgttcatc ttcatcacaa ccttcatggt ttgcgcccgg tcgtccatca tggccttaag
     1081 ttcttccagc cattggtcac gctcgcgcaa aatgccgtgg cgctccatgt gggctcgtgg
     1141 tgtcaataca cgagtattgt caggggtgta cgttccgcct tggaaacgct cggtgatgcg
     1201 atcaacgtca accagctcag ggaggccggc cagcggtgcg ccggtgattt cgcagtgtga
     1261 tgcttgctct ttgagtactt gcttgttcat gggtgctact cccaataaaa aaatcagcgc
     1321 cctcgatccg gtagcagttg gcggggcaag atccgcctcg gataaaggac actgacgttt
     1381 ttatcctgcc tttcaatttg gcctgctaca gcctacggat aactataatg cgtgtttacg
     1441 ggcgtgtcaa acgaaaaaag ccccgaccgt agccggggct ccctgttacg ccaccaatca
     1501 gcccagaagc aggccggtat ggcgcggttg tgtgactttg acgccccacg caagtccgac
     1561 ctcgtaccgc actttgcgat agccgggata atggctgctg cccatggtat atctccttat
     1621 taaagttaaa caaaattatt tctacagggg aattgttatc cgctcacaat tcccctatag
     1681 tgagtcgtat taatttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac
     1741 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     1801 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     1861 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     1921 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     1981 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     2041 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     2101 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     2161 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     2221 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     2281 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     2341 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     2401 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     2461 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     2521 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     2581 gccattaccg agtccgggct gcgcgttggt gcggacatct cggtagtggg atacgacgat
     2641 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     2701 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     2761 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     2821 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     2881 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc
     2941 gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     3001 gggcatgact aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca
     3061 tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct
     3121 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     3181 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     3241 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     3301 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     3361 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     3421 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     3481 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     3541 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     3601 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     3661 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     3721 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     3781 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     3841 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     3901 ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg cttttgccgt
     3961 tacgcaccac cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg
     4021 gagcacctca aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca
     4081 ctttgcgccg aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt
     4141 gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg
     4201 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     4261 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     4321 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     4381 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     4441 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     4501 gctcatccgg agttccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     4561 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     4621 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     4681 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     4741 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     4801 cccgttttca ctatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     4861 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     4921 caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
     4981 ccttaaacgc ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat
     5041 tcgaaagcaa attcgacccg gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg
     5101 tctattgctg gtttaccggt ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa
     5161 atgcctgagg tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     5221 ctagttattg ctcagcggtg gcagcagcct agtccgctgc gcttactgag ccgtagaatt
     5281 ctgccattgc tcgctgtcct cagtcttgcg cttggggcgg cccggtgagc gcttgggctt
     5341 ttcgttggga tcagtctctc gcttcgggtc aaagtcggct ttgttgatgc gcaccgtttg
     5401 gccgtttcgc tcaaccagca tggtttcggg gtgttttggc atggttgcct cctacattgc
     5461 cttggtcaat tgtaccacac gatcagacgt aaaaaagccc gctctgtggc gggctcgctc
     5521 ggaacagttg gctgccttgt agaacttggc tcgcttaacc gaattggctg tcttgtacc
//
