LOCUS       sequence-330741-        9817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of PmcCAST TnsAB, TnsC, TniQ and TnsD 
            proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9817)
  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 9817)
  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: pMS25: pTns(PmcCAST)
FEATURES             Location/Qualifiers
     source          1..9817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(371..1453)
                     /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     1415..1434
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1454..1531)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1598..1616
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1617..1641
                     /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        6965..6983
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6984..7008
                     /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(8062..8080)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8076..8123
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(8244..9035)
                     /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(9036..9127)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaccttcga tcaaaccacc tccccaggtg gttttttcga aggttaagtc agttggggaa
       61 ctgcttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga
      121 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct
      181 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
      241 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
      301 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
      361 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      421 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
      481 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
      541 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
      601 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
      661 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
      721 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
      781 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
      841 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
      901 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
      961 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     1021 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     1081 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     1141 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     1201 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     1261 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     1321 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     1381 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     1441 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     1501 cgcgaaaggt tttgcgccat tcgatggtgt caagcggcta tttaacgacc ctcgggatct
     1561 cgacgctctc ccttatgcga ctcctgcatt aggaaattaa tacgactcac tataggggaa
     1621 ttgtgagcgg ataacaattc ccctgtagaa ataattttgt ttaactttaa taaggagata
     1681 taccatgaaa gagcaatttg aacaatgggt tgctcgcctt caaatatccg aggcagggca
     1741 aaaaattatt gagaaagttc gctcatctga gccatcccgt cgcgttggtg gcggtagcaa
     1801 gaatgtttcg ggacgctacc ctagtcgcaa gatgggagtc acgattcaat ttgagtctca
     1861 tcgggttgaa ctgccaatta tctatcagtt agaacatgat aaagatgttc tagaattcta
     1921 cgatcaacca ccgcaaatta aacttgacta tcaagcaagt aacggacgta gattaggcgt
     1981 tttacacacc ccagatttat ttgtcatccg aaccaattcg gctggttggg aagagtgtaa
     2041 aactgagcag gatttgaaaa aactagcaga gaaaaacccc aatcgttact tttactctga
     2101 agaaaataat caatggcatt ccttgccagg agaagtttat gctaaccagt ttgggcttta
     2161 tcaccgcatt cattcagatc gcgaaatcaa ttgggttctg caacgtaatt tgcaattttt
     2221 agaagattat tatcgctcag aatctctagt agtggaagaa gcgatcgctc aatcattact
     2281 tgcaattgtg tcatctcaac caggaatcac tttagcagaa ttactcaatc ggtcaacagg
     2341 agctaaatct gacgatatct acaccttaat tattcaagag caaatctata ttgatttaaa
     2401 tgctacacca ctagctgaac cagaaagatg tctaatcttc cgtgacgagc aaactgctag
     2461 tgcttataga ttaatggttg agcaaccaag tgtaagtctt ccggctatct ctcctgtagt
     2521 caatattgtt actggagcaa ctgttagttg ggacggcaaa ggtttaaata tcatccatgt
     2581 tggagaaaca gaggttatac ttggcgcaga aaataatcag ttaattgaac tcaaaaaagc
     2641 tatatttgag aatctggttc gccaaggaaa aatcacaagt ctccaaacac cagaaaaaac
     2701 agctattagc acagaatctt ggcagcgatt tcatcaagct agtcctgaag accaagcaga
     2761 agcccttgaa cgctacaaaa ctatcgaacc ttacttaaac ggacatcctc cagaaaacga
     2821 aacaatacca gcccgtacca ttcgtcactg gaaagctaaa tatttaattg caattcaaaa
     2881 atacggttgc ggctacattg ggcttttgtc ccatcgcagt gtcaaaggta atcgacagcg
     2941 caagctgccc gaagacacct tagcaattat ggagaagttt attttagaag actatgaaac
     3001 tcttaaacaa aaaagaatgt gggaggttca cgcggcttta gttcgagctt gcgaagaagc
     3061 tggtgtaatt gctccgagct ataaagcatt taccaaagaa gtacaacgcc ggactggtta
     3121 cgaacagact aaaaaacgcc aaggtcgtcg tgccgcttat cagcatgaat ccttctattg
     3181 ggagcttgct ataactaccc caagacacgg cgatcgccct tttgaaattg ggcatataga
     3241 tcatactgag ttagatgtag agctagtttg ctctgataca ggtcgcaatc ttggcagacc
     3301 ttgggcaaca tttctcgtag atgcttattc acggcgattg ctggcagttt acctaacgtt
     3361 tgactcaccc tcatatcgtt cttgcttaat ggtgctaaga atttgtgtca agcgtcatgg
     3421 caggcttccc caaattgtag tggtagataa cggtgcagag tttcacagcg tttactttga
     3481 aacactacta gctacttttg aatgtacaaa aaagcaaaga cccccagcca aagctcgttt
     3541 tggttcggtt tgcgaacgcc tatttggaac ttccaatact caatttgtcc ataacttact
     3601 tggtaataca caaatcactc gtaatgtccg ccaagtcact aagtcagtta accctaaaaa
     3661 tctagctgta tggacacttg gtttacttta tgaatatctt tgtgcttggg cttacgaagt
     3721 ttatgataca gacgaacatc cggcactttt ccagagtccg agggatgcgt ttgctgcggg
     3781 tatggcgatt ggaggcagtc gggttcaccg catgattcct tacgatgaaa actttcagat
     3841 tcttacccta cctacaacct cagagggaaa agcaaaagtt caagttggac gcggcataaa
     3901 aattaactac atatactact ggtctaatag tttccgcgat cctcaaattg aaaatacctc
     3961 tgtgcagata agatacgacc cgtttaatat tggcatcgct tacgcctttg ttcgaggaca
     4021 gtgggtacag tgtatctctc aatattatgc agaactccaa ggtcgttcag aaaaagaact
     4081 gaagttagct agtatcgagt tacgcaaaca ctattcaaat catacacaac aatctaaagt
     4141 ttcggctaag aatttagcag aatttttagc atcggtagaa gctcaagaag ctctattaga
     4201 acagcgttcc tatgatgcag aagttaagga agtctttagg gtaatcgagg gtggaaaagc
     4261 gacaacaagc cgtaacgaag aaccaaaact tattcaagtt acttttgcca ataccgattt
     4321 tcaagcggat gaagatgaag cgatcgtccc agaaacactt gtagtgtatg aggagtttta
     4381 actatgacaa aatctactgg atttccgctt gaactcctca cacggcctgc tacagaaagg
     4441 cttgcttact ttgaaaatta caccgtagcg catcctcgac ttaaagaagt ctatgaaatt
     4501 ctcatgcgaa cgattgccga accagcagga gcttcattta tctttgttta cggtgctagt
     4561 ggtgttggta aaacaacttt gcgactccgc gttgagcaga aattaacaga actagcatta
     4621 cctaaattgg aaagcgatcg cgctcgtgtt cctgttgttg gtattgaagc tatagcgcca
     4681 gaatcgagat acttcaactg gaaagaatac tacacacgcg ccttaattac tttggaagaa
     4741 ccattaatcg atcacaagtt tgattacggt gtgcgtggta tctctcgcga taattttggc
     4801 aaaattaatg ttgagtcgaa agtagttgca cctgctctac gcagagcttt agaaaatgct
     4861 ctcatccatc gtcatccaga cgtatttttt gtagatgaag cacaacattt tggtaaggtg
     4921 gcaagcggat acaaactaca agatcaatta gactgtctca aatctttagc aaatatgaca
     4981 ggcatcttgc actgtctatt aggaacatac gaactactta cattccgtaa tttgagcggt
     5041 caactctcac gtcgtagtgt tgatattcat ttccgccgtt attgtgctga tagcccagaa
     5101 gatgtacagg cttttaagag tgtgttgtta acttttcaac agcatctgcc actagcagaa
     5161 actccaaatt tagtcgatca ttgggaatat ttttacgaac gtactttagg atgtattggc
     5221 acactgaaag attggctgaa gcgtgttctt agtgatgctt tggatcgaga agcaacaaca
     5281 atcactctca aagatttaca aaaacgtgct ttgtcggttg ctcagtgtca gaaaatgttc
     5341 aaggaaattc aagagggtga acgacaatta tctgaaacag aagcagatgt gcaaaactta
     5401 cgctctgcat taggacttgg cgcaaaacca attgtgctac ccgaagaaac cccaaaaact
     5461 actcgtccgc caggaaaggt tggcaaacgc aaacctaaaa gagatcccat cggagtgcag
     5521 caagatgtta gctagtgtat tagaaacata cgaatcttgg aatttaaaaa agccattaat
     5581 tccacaacgt agtcgcctct atcaacctca acctgtaggt attgggactc cttacataga
     5641 gagtttaaca ggttatatca cacggatagc agagttgcat ggagttcttc caggagtctt
     5701 gatgactaga gaaattgctc cgcttgtcaa taaaatatat ttccaaaatg gagctaatcg
     5761 aggttttcgt gaaatattca accgtagtca agcgttaaat ggcatgggag agatggcagc
     5821 agatttagtt caagtgttgc aaaaattgac gcttagagat gacttgcgtt ttttaaccat
     5881 gctgttttgg tctaacattt tgactccacg caatttattt cgtaccagaa aagcttggtg
     5941 tccaatttgt tatcaagagc ggcatcaaaa tgggctggta gtctatgagc aattgctttg
     6001 gacaattaac ctaattacaa tttgcccgca gcatcaaaaa cccttggttg aactttgccc
     6061 tcactgtaat catgaatcac ctttgttaaa ctggcgctca agacctggtt actgttcaaa
     6121 gtgtggagag tggttaggcg caaatcaatg ccttaagaca tttacagatg gtgaaggatc
     6181 aataaaatta cagttggagt ggcagtattg gactgccaat gttgtaggag agctaatttt
     6241 agcttctcaa tgctttgaat ctgccccatc aaaagaaaat attacaaaat cattaaatat
     6301 agttattgat aaagttgcgg aaaataatgc agctgcattt tctcgtctta ttggagtacc
     6361 aaaaaatagt ttgtggatgt ggcagtctac taaaaccttg ccagagctaa atacacttct
     6421 gaaaatttgt tacgaattag agatatcgtt ggtagaattt cttactccaa aaaatctaat
     6481 tactaaatct tttacaaaaa tctcgcagaa gcacctccaa ctttctcgca caccaagggt
     6541 atctccaaaa tctttcgacc aatatcaagt aaaagatgct ttattagcaa tacttgcagg
     6601 taatgaagaa ccaccaccaa caatggaaga agttggaaag cgtttaggtc atcacaacag
     6661 aactatttct agacactttc cagatttgtg tagtgctatt tctgctaaat gtcgtaatta
     6721 caataaagct tgccgtctca aaagcataga aaaactttgc agcgaagtta gagagattgt
     6781 ccttagcctt aatgctcagg gtgtgtatcc cacagaaggt cgtgtttgtg aattaatgcc
     6841 aaatccgggg tgttttagat ataaacaagt ccgcgctgct tttaatgatg cacgacgtga
     6901 gtttggtttg taacttaagt cgaacagaaa gtaatcgtat tgtacacggc cgcataatcg
     6961 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc atcttagtat
     7021 attagttaag tataagaagg agatatacat gactatttac tcagaacatt ggagcctaaa
     7081 tcctctagaa atcccgcaac ggagtcggtt attttctctt gaacctgtgg cagtgggaac
     7141 tccctatgca gaaagtctta gtagctatct ccatcgttta gcgcaggcgc attgtttaac
     7201 ctctgagaag ctcgttatgg gagaaattgc gccacttatt ctcaaagatg aggataaatc
     7261 tgagctattg tcaaaaaatc tgagtcacct attgggaaat agcgacgcta aaccggctat
     7321 taacggaatg cgagaaatga cagaaaagtt agtcactgtc ttagaagagt taacaatgcg
     7381 tcaggattta cgctttttga cccttttaag ctggaaagga atgatttatg acaagggatt
     7441 atttcggaac tatcgcgctt ggtgtccttg ttgttgtgaa gagtggatgc agaagaacaa
     7501 aactatctat gaaccattat catggtcatt taaggacgta gaattttgtt tgattcacaa
     7561 acagcgatta atagaagaat gttctcattg tggtgcccgt ttacctgtga tggctaggct
     7621 ttcacctgcg ggattttgtt ctcgttgtta cgggtggtta ggacaggaaa ttaaaggcga
     7681 agaagaaata gagaaatatc gagttaatat tcaaggaatt agcgaattaa ttgcacttac
     7741 tccccaatta ggttataaac ccatcccgat cgagttaacc cgaaaattac aattgatttt
     7801 gttagtgttt gaacaagcaa ttggaaagga tgtgaagtta ttgggagatt taggaggaat
     7861 catggaatcg ttgaggatag cctcaacaac aaatcaaagt caaccttatc atttggtgaa
     7921 gttgattatt cctgtgtgcg agaaggcgaa gattagtgta ttccagttgt ttgggtcaga
     7981 ttttaaggaa ttgggcaaga tattgttcgg gaattttagt ctggaattaa agttatagtt
     8041 aattaaccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     8101 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
     8161 tgcttccggt agtgccctga accgacgacc gggtcatcgt ggccggatct tgcggcccct
     8221 cggcttgaac gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg
     8281 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca
     8341 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt
     8401 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg
     8461 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat
     8521 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt
     8581 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata
     8641 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc
     8701 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc
     8761 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt
     8821 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta
     8881 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa
     8941 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt
     9001 tgagtcgata cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga
     9061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9121 aaacaaatag ctagctcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg
     9181 cggacacata caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc
     9241 aaaacagcag ggccgcgccg gtggcgtttt tccataggct ccgccctcct gccagagttc
     9301 acataaacag acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg
     9361 acagtacggg cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc
     9421 ctcttgcgct ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc
     9481 ttacgggaag tgtggcgctt tctcatagct cacacactgg tatctcggct cggtgtaggt
     9541 cgttcgctcc aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt
     9601 atccggtaac tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc
     9661 agccattggt aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg
     9721 aagtgtgcgc caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa
     9781 agccagttac cacggttaag cagttcccca actgact
//
