LOCUS       sequence-330721-        8439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of PmcCAST TnsAB, TnsC and TniQ proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8439)
  AUTHORS   Saito M, Ladha A, Strecker J, Faure G, Neumann E, Altae-Tran H, 
            Macrae RK, Zhang F
  TITLE     Dual modes of CRISPR-associated transposon homing.
  JOURNAL   Cell. 2021 Apr 29;184(9):2441-2453.e18. doi: 
            10.1016/j.cell.2021.03.006. Epub 2021 Mar 25.
  PUBMED    33770501
REFERENCE   2  (bases 1 to 8439)
  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: 
            "10.1016/j.cell.2021.03.006"; journalName: "Cell"; date: 
            "2021-04-29- 29"; volume: "184"; issue: "9"; pages: "2441-2453.e18"
COMMENT     Sequence Label: pMS27: pTns(PmcCAST)_Delta-TnsD
FEATURES             Location/Qualifiers
     source          1..8439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(250..1332)
                     /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     1294..1313
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1333..1410)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1477..1495
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1496..1520
                     /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)."
     promoter        5466..5484
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5485..5509
                     /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(6563..6581)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6577..6624
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6745..7536)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(7537..7628)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta
       61 gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa
      121 gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag
      181 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
      241 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
      301 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
      361 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
      421 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
      481 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc
      541 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
      601 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
      661 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
      721 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
      781 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
      841 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
      901 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
      961 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     1021 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     1081 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     1141 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     1201 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     1261 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     1321 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     1381 gcgaaaggtt ttgcgccatt cgatggtgtc aagcggctat ttaacgaccc tcgggatctc
     1441 gacgctctcc cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat
     1501 tgtgagcgga taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat
     1561 accatgaaag agcaatttga acaatgggtt gctcgccttc aaatatccga ggcagggcaa
     1621 aaaattattg agaaagttcg ctcatctgag ccatcccgtc gcgttggtgg cggtagcaag
     1681 aatgtttcgg gacgctaccc tagtcgcaag atgggagtca cgattcaatt tgagtctcat
     1741 cgggttgaac tgccaattat ctatcagtta gaacatgata aagatgttct agaattctac
     1801 gatcaaccac cgcaaattaa acttgactat caagcaagta acggacgtag attaggcgtt
     1861 ttacacaccc cagatttatt tgtcatccga accaattcgg ctggttggga agagtgtaaa
     1921 actgagcagg atttgaaaaa actagcagag aaaaacccca atcgttactt ttactctgaa
     1981 gaaaataatc aatggcattc cttgccagga gaagtttatg ctaaccagtt tgggctttat
     2041 caccgcattc attcagatcg cgaaatcaat tgggttctgc aacgtaattt gcaattttta
     2101 gaagattatt atcgctcaga atctctagta gtggaagaag cgatcgctca atcattactt
     2161 gcaattgtgt catctcaacc aggaatcact ttagcagaat tactcaatcg gtcaacagga
     2221 gctaaatctg acgatatcta caccttaatt attcaagagc aaatctatat tgatttaaat
     2281 gctacaccac tagctgaacc agaaagatgt ctaatcttcc gtgacgagca aactgctagt
     2341 gcttatagat taatggttga gcaaccaagt gtaagtcttc cggctatctc tcctgtagtc
     2401 aatattgtta ctggagcaac tgttagttgg gacggcaaag gtttaaatat catccatgtt
     2461 ggagaaacag aggttatact tggcgcagaa aataatcagt taattgaact caaaaaagct
     2521 atatttgaga atctggttcg ccaaggaaaa atcacaagtc tccaaacacc agaaaaaaca
     2581 gctattagca cagaatcttg gcagcgattt catcaagcta gtcctgaaga ccaagcagaa
     2641 gcccttgaac gctacaaaac tatcgaacct tacttaaacg gacatcctcc agaaaacgaa
     2701 acaataccag cccgtaccat tcgtcactgg aaagctaaat atttaattgc aattcaaaaa
     2761 tacggttgcg gctacattgg gcttttgtcc catcgcagtg tcaaaggtaa tcgacagcgc
     2821 aagctgcccg aagacacctt agcaattatg gagaagttta ttttagaaga ctatgaaact
     2881 cttaaacaaa aaagaatgtg ggaggttcac gcggctttag ttcgagcttg cgaagaagct
     2941 ggtgtaattg ctccgagcta taaagcattt accaaagaag tacaacgccg gactggttac
     3001 gaacagacta aaaaacgcca aggtcgtcgt gccgcttatc agcatgaatc cttctattgg
     3061 gagcttgcta taactacccc aagacacggc gatcgccctt ttgaaattgg gcatatagat
     3121 catactgagt tagatgtaga gctagtttgc tctgatacag gtcgcaatct tggcagacct
     3181 tgggcaacat ttctcgtaga tgcttattca cggcgattgc tggcagttta cctaacgttt
     3241 gactcaccct catatcgttc ttgcttaatg gtgctaagaa tttgtgtcaa gcgtcatggc
     3301 aggcttcccc aaattgtagt ggtagataac ggtgcagagt ttcacagcgt ttactttgaa
     3361 acactactag ctacttttga atgtacaaaa aagcaaagac ccccagccaa agctcgtttt
     3421 ggttcggttt gcgaacgcct atttggaact tccaatactc aatttgtcca taacttactt
     3481 ggtaatacac aaatcactcg taatgtccgc caagtcacta agtcagttaa ccctaaaaat
     3541 ctagctgtat ggacacttgg tttactttat gaatatcttt gtgcttgggc ttacgaagtt
     3601 tatgatacag acgaacatcc ggcacttttc cagagtccga gggatgcgtt tgctgcgggt
     3661 atggcgattg gaggcagtcg ggttcaccgc atgattcctt acgatgaaaa ctttcagatt
     3721 cttaccctac ctacaacctc agagggaaaa gcaaaagttc aagttggacg cggcataaaa
     3781 attaactaca tatactactg gtctaatagt ttccgcgatc ctcaaattga aaatacctct
     3841 gtgcagataa gatacgaccc gtttaatatt ggcatcgctt acgcctttgt tcgaggacag
     3901 tgggtacagt gtatctctca atattatgca gaactccaag gtcgttcaga aaaagaactg
     3961 aagttagcta gtatcgagtt acgcaaacac tattcaaatc atacacaaca atctaaagtt
     4021 tcggctaaga atttagcaga atttttagca tcggtagaag ctcaagaagc tctattagaa
     4081 cagcgttcct atgatgcaga agttaaggaa gtctttaggg taatcgaggg tggaaaagcg
     4141 acaacaagcc gtaacgaaga accaaaactt attcaagtta cttttgccaa taccgatttt
     4201 caagcggatg aagatgaagc gatcgtccca gaaacacttg tagtgtatga ggagttttaa
     4261 ctatgacaaa atctactgga tttccgcttg aactcctcac acggcctgct acagaaaggc
     4321 ttgcttactt tgaaaattac accgtagcgc atcctcgact taaagaagtc tatgaaattc
     4381 tcatgcgaac gattgccgaa ccagcaggag cttcatttat ctttgtttac ggtgctagtg
     4441 gtgttggtaa aacaactttg cgactccgcg ttgagcagaa attaacagaa ctagcattac
     4501 ctaaattgga aagcgatcgc gctcgtgttc ctgttgttgg tattgaagct atagcgccag
     4561 aatcgagata cttcaactgg aaagaatact acacacgcgc cttaattact ttggaagaac
     4621 cattaatcga tcacaagttt gattacggtg tgcgtggtat ctctcgcgat aattttggca
     4681 aaattaatgt tgagtcgaaa gtagttgcac ctgctctacg cagagcttta gaaaatgctc
     4741 tcatccatcg tcatccagac gtattttttg tagatgaagc acaacatttt ggtaaggtgg
     4801 caagcggata caaactacaa gatcaattag actgtctcaa atctttagca aatatgacag
     4861 gcatcttgca ctgtctatta ggaacatacg aactacttac attccgtaat ttgagcggtc
     4921 aactctcacg tcgtagtgtt gatattcatt tccgccgtta ttgtgctgat agcccagaag
     4981 atgtacaggc ttttaagagt gtgttgttaa cttttcaaca gcatctgcca ctagcagaaa
     5041 ctccaaattt agtcgatcat tgggaatatt tttacgaacg tactttagga tgtattggca
     5101 cactgaaaga ttggctgaag cgtgttctta gtgatgcttt ggatcgagaa gcaacaacaa
     5161 tcactctcaa agatttacaa aaacgtgctt tgtcggttgc tcagtgtcag aaaatgttca
     5221 aggaaattca agagggtgaa cgacaattat ctgaaacaga agcagatgtg caaaacttac
     5281 gctctgcatt aggacttggc gcaaaaccaa ttgtgctacc cgaagaaacc ccaaaaacta
     5341 ctcgtccgcc aggaaaggtt ggcaaacgca aacctaaaag agatcccatc ggagtgcagc
     5401 aagatgttag ctagcttaag tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc
     5461 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc catcttagta
     5521 tattagttaa gtataagaag gagatataca tgactattta ctcagaacat tggagcctaa
     5581 atcctctaga aatcccgcaa cggagtcggt tattttctct tgaacctgtg gcagtgggaa
     5641 ctccctatgc agaaagtctt agtagctatc tccatcgttt agcgcaggcg cattgtttaa
     5701 cctctgagaa gctcgttatg ggagaaattg cgccacttat tctcaaagat gaggataaat
     5761 ctgagctatt gtcaaaaaat ctgagtcacc tattgggaaa tagcgacgct aaaccggcta
     5821 ttaacggaat gcgagaaatg acagaaaagt tagtcactgt cttagaagag ttaacaatgc
     5881 gtcaggattt acgctttttg acccttttaa gctggaaagg aatgatttat gacaagggat
     5941 tatttcggaa ctatcgcgct tggtgtcctt gttgttgtga agagtggatg cagaagaaca
     6001 aaactatcta tgaaccatta tcatggtcat ttaaggacgt agaattttgt ttgattcaca
     6061 aacagcgatt aatagaagaa tgttctcatt gtggtgcccg tttacctgtg atggctaggc
     6121 tttcacctgc gggattttgt tctcgttgtt acgggtggtt aggacaggaa attaaaggcg
     6181 aagaagaaat agagaaatat cgagttaata ttcaaggaat tagcgaatta attgcactta
     6241 ctccccaatt aggttataaa cccatcccga tcgagttaac ccgaaaatta caattgattt
     6301 tgttagtgtt tgaacaagca attggaaagg atgtgaagtt attgggagat ttaggaggaa
     6361 tcatggaatc gttgaggata gcctcaacaa caaatcaaag tcaaccttat catttggtga
     6421 agttgattat tcctgtgtgc gagaaggcga agattagtgt attccagttg tttgggtcag
     6481 attttaagga attgggcaag atattgttcg ggaattttag tctggaatta aagttatagt
     6541 taattaacct aggctgctgc caccgctgag caataactag cataacccct tggggcctct
     6601 aaacgggtct tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca
     6661 ctgcttccgg tagtgccctg aaccgacgac cgggtcatcg tggccggatc ttgcggcccc
     6721 tcggcttgaa cgaattgtta gacattattt gccgactacc ttggtgatct cgcctttcac
     6781 gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc
     6841 aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca ggcgctccat
     6901 tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc tgtaccaaat
     6961 gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg gcgagttcca
     7021 tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag
     7081 ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg tcagcaagat
     7141 agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt cattgcgctg
     7201 ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg
     7261 cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg aagccgaagt
     7321 ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta cggtcaccgt
     7381 aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg agccgtacaa
     7441 atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta cctctgatag
     7501 ttgagtcgat acttcggcga tcaccgcttc cctcatactc ttcctttttc aatattattg
     7561 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7621 taaacaaata gctagctcac tcggtcgcta cgctccgggc gtgagactgc ggcgggcgct
     7681 gcggacacat acaaagttac ccacagattc cgtggataag caggggacta acatgtgagg
     7741 caaaacagca gggccgcgcc ggtggcgttt ttccataggc tccgccctcc tgccagagtt
     7801 cacataaaca gacgcttttc cggtgcatct gtgggagccg tgaggctcaa ccatgaatct
     7861 gacagtacgg gcgaaacccg acaggactta aagatcccca ccgtttccgg cgggtcgctc
     7921 cctcttgcgc tctcctgttc cgaccctgcc gtttaccgga tacctgttcc gcctttctcc
     7981 cttacgggaa gtgtggcgct ttctcatagc tcacacactg gtatctcggc tcggtgtagg
     8041 tcgttcgctc caagctgggc tgtaagcaag aactccccgt tcagcccgac tgctgcgcct
     8101 tatccggtaa ctgttcactt gagtccaacc cggaaaagca cggtaaaacg ccactggcag
     8161 cagccattgg taactgggag ttcgcagagg atttgtttag ctaaacacgc ggttgctctt
     8221 gaagtgtgcg ccaaagtccg gctacactgg aaggacagat ttggttgctg tgctctgcga
     8281 aagccagtta ccacggttaa gcagttcccc aactgactta accttcgatc aaaccacctc
     8341 cccaggtggt tttttcgaag gttaagtcag ttggggaact gcttaacctt cgatcaaacc
     8401 acctccccag gtggtttttt cgtttacagg gcaaaagat
//
