LOCUS       sequence-330385-        8954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of PmcCAST TnsAB, TnsC and TnsD proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8954)
  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 8954)
  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: pMS26: pTns(PmcCAST)_Delta-TniQ
FEATURES             Location/Qualifiers
     source          1..8954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        794..885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             886..1677
                     /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"
     terminator      complement(1798..1845)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1841..1859
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1897..1941)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    2050..2074
                     /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        complement(2075..2093)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7417..7441
                     /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        complement(7442..7460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        7527..7604
                     /gene="lacI"
                     /label=lacI promoter
     CDS             7605..8687
                     /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(7624..7643)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gaaaaaacca cctggggagg tggtttgatc gaaggttaag cagttcccca actgacttaa
       61 ccttcgaaaa aaccacctgg ggaggtggtt tgatcgaagg ttaagtcagt tggggaactg
      121 cttaaccgtg gtaactggct ttcgcagagc acagcaacca aatctgtcct tccagtgtag
      181 ccggactttg gcgcacactt caagagcaac cgcgtgttta gctaaacaaa tcctctgcga
      241 actcccagtt accaatggct gctgccagtg gcgttttacc gtgcttttcc gggttggact
      301 caagtgaaca gttaccggat aaggcgcagc agtcgggctg aacggggagt tcttgcttac
      361 agcccagctt ggagcgaacg acctacaccg agccgagata ccagtgtgtg agctatgaga
      421 aagcgccaca cttcccgtaa gggagaaagg cggaacaggt atccggtaaa cggcagggtc
      481 ggaacaggag agcgcaagag ggagcgaccc gccggaaacg gtggggatct ttaagtcctg
      541 tcgggtttcg cccgtactgt cagattcatg gttgagcctc acggctccca cagatgcacc
      601 ggaaaagcgt ctgtttatgt gaactctggc aggagggcgg agcctatgga aaaacgccac
      661 cggcgcggcc ctgctgtttt gcctcacatg ttagtcccct gcttatccac ggaatctgtg
      721 ggtaactttg tatgtgtccg cagcgcccgc cgcagtctca cgcccggagc gtagcgaccg
      781 agtgagctag ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      841 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag ggaagcggtg
      901 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
      961 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     1021 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     1081 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     1141 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     1201 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     1261 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     1321 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     1381 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     1441 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     1501 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     1561 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     1621 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataatgt
     1681 ctaacaattc gttcaagccg aggggccgca agatccggcc acgatgaccc ggtcgtcggt
     1741 tcagggcact accggaagca gtgtgaccgt gtgcttctca aatgcctgag gtttcagcaa
     1801 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt
     1861 ggcagcagcc taggttaatt aagctgcgct agtagacgag tccatgtgct ggcgttcaaa
     1921 tttcgcagca gcggtttctt taccagactc gagggtaccg acgtcagcga tcgcgtggcc
     1981 ggccgatatc caattgagat ctgccatatg tatatctcct tcttatactt aactaatata
     2041 ctaagatggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcga
     2101 ttatgcggcc gtgtacaata cgattacttt ctgttcgact taagttacaa accaaactca
     2161 cgtcgtgcat cattaaaagc agcgcggact tgtttatatc taaaacaccc cggatttggc
     2221 attaattcac aaacacgacc ttctgtggga tacacaccct gagcattaag gctaaggaca
     2281 atctctctaa cttcgctgca aagtttttct atgcttttga gacggcaagc tttattgtaa
     2341 ttacgacatt tagcagaaat agcactacac aaatctggaa agtgtctaga aatagttctg
     2401 ttgtgatgac ctaaacgctt tccaacttct tccattgttg gtggtggttc ttcattacct
     2461 gcaagtattg ctaataaagc atcttttact tgatattggt cgaaagattt tggagatacc
     2521 cttggtgtgc gagaaagttg gaggtgcttc tgcgagattt ttgtaaaaga tttagtaatt
     2581 agattttttg gagtaagaaa ttctaccaac gatatctcta attcgtaaca aattttcaga
     2641 agtgtattta gctctggcaa ggttttagta gactgccaca tccacaaact attttttggt
     2701 actccaataa gacgagaaaa tgcagctgca ttattttccg caactttatc aataactata
     2761 tttaatgatt ttgtaatatt ttcttttgat ggggcagatt caaagcattg agaagctaaa
     2821 attagctctc ctacaacatt ggcagtccaa tactgccact ccaactgtaa ttttattgat
     2881 ccttcaccat ctgtaaatgt cttaaggcat tgatttgcgc ctaaccactc tccacacttt
     2941 gaacagtaac caggtcttga gcgccagttt aacaaaggtg attcatgatt acagtgaggg
     3001 caaagttcaa ccaagggttt ttgatgctgc gggcaaattg taattaggtt aattgtccaa
     3061 agcaattgct catagactac cagcccattt tgatgccgct cttgataaca aattggacac
     3121 caagcttttc tggtacgaaa taaattgcgt ggagtcaaaa tgttagacca aaacagcatg
     3181 gttaaaaaac gcaagtcatc tctaagcgtc aatttttgca acacttgaac taaatctgct
     3241 gccatctctc ccatgccatt taacgcttga ctacggttga atatttcacg aaaacctcga
     3301 ttagctccat tttggaaata tattttattg acaagcggag caatttctct agtcatcaag
     3361 actcctggaa gaactccatg caactctgct atccgtgtga tataacctgt taaactctct
     3421 atgtaaggag tcccaatacc tacaggttga ggttgataga ggcgactacg ttgtggaatt
     3481 aatggctttt ttaaattcca agattcgtat gtttctaata cactagctaa catcttgctg
     3541 cactccgatg ggatctcttt taggtttgcg tttgccaacc tttcctggcg gacgagtagt
     3601 ttttggggtt tcttcgggta gcacaattgg ttttgcgcca agtcctaatg cagagcgtaa
     3661 gttttgcaca tctgcttctg tttcagataa ttgtcgttca ccctcttgaa tttccttgaa
     3721 cattttctga cactgagcaa ccgacaaagc acgtttttgt aaatctttga gagtgattgt
     3781 tgttgcttct cgatccaaag catcactaag aacacgcttc agccaatctt tcagtgtgcc
     3841 aatacatcct aaagtacgtt cgtaaaaata ttcccaatga tcgactaaat ttggagtttc
     3901 tgctagtggc agatgctgtt gaaaagttaa caacacactc ttaaaagcct gtacatcttc
     3961 tgggctatca gcacaataac ggcggaaatg aatatcaaca ctacgacgtg agagttgacc
     4021 gctcaaatta cggaatgtaa gtagttcgta tgttcctaat agacagtgca agatgcctgt
     4081 catatttgct aaagatttga gacagtctaa ttgatcttgt agtttgtatc cgcttgccac
     4141 cttaccaaaa tgttgtgctt catctacaaa aaatacgtct ggatgacgat ggatgagagc
     4201 attttctaaa gctctgcgta gagcaggtgc aactactttc gactcaacat taattttgcc
     4261 aaaattatcg cgagagatac cacgcacacc gtaatcaaac ttgtgatcga ttaatggttc
     4321 ttccaaagta attaaggcgc gtgtgtagta ttctttccag ttgaagtatc tcgattctgg
     4381 cgctatagct tcaataccaa caacaggaac acgagcgcga tcgctttcca atttaggtaa
     4441 tgctagttct gttaatttct gctcaacgcg gagtcgcaaa gttgttttac caacaccact
     4501 agcaccgtaa acaaagataa atgaagctcc tgctggttcg gcaatcgttc gcatgagaat
     4561 ttcatagact tctttaagtc gaggatgcgc tacggtgtaa ttttcaaagt aagcaagcct
     4621 ttctgtagca ggccgtgtga ggagttcaag cggaaatcca gtagattttg tcatagttaa
     4681 aactcctcat acactacaag tgtttctggg acgatcgctt catcttcatc cgcttgaaaa
     4741 tcggtattgg caaaagtaac ttgaataagt tttggttctt cgttacggct tgttgtcgct
     4801 tttccaccct cgattaccct aaagacttcc ttaacttctg catcatagga acgctgttct
     4861 aatagagctt cttgagcttc taccgatgct aaaaattctg ctaaattctt agccgaaact
     4921 ttagattgtt gtgtatgatt tgaatagtgt ttgcgtaact cgatactagc taacttcagt
     4981 tctttttctg aacgaccttg gagttctgca taatattgag agatacactg tacccactgt
     5041 cctcgaacaa aggcgtaagc gatgccaata ttaaacgggt cgtatcttat ctgcacagag
     5101 gtattttcaa tttgaggatc gcggaaacta ttagaccagt agtatatgta gttaattttt
     5161 atgccgcgtc caacttgaac ttttgctttt ccctctgagg ttgtaggtag ggtaagaatc
     5221 tgaaagtttt catcgtaagg aatcatgcgg tgaacccgac tgcctccaat cgccataccc
     5281 gcagcaaacg catccctcgg actctggaaa agtgccggat gttcgtctgt atcataaact
     5341 tcgtaagccc aagcacaaag atattcataa agtaaaccaa gtgtccatac agctagattt
     5401 ttagggttaa ctgacttagt gacttggcgg acattacgag tgatttgtgt attaccaagt
     5461 aagttatgga caaattgagt attggaagtt ccaaataggc gttcgcaaac cgaaccaaaa
     5521 cgagctttgg ctgggggtct ttgctttttt gtacattcaa aagtagctag tagtgtttca
     5581 aagtaaacgc tgtgaaactc tgcaccgtta tctaccacta caatttgggg aagcctgcca
     5641 tgacgcttga cacaaattct tagcaccatt aagcaagaac gatatgaggg tgagtcaaac
     5701 gttaggtaaa ctgccagcaa tcgccgtgaa taagcatcta cgagaaatgt tgcccaaggt
     5761 ctgccaagat tgcgacctgt atcagagcaa actagctcta catctaactc agtatgatct
     5821 atatgcccaa tttcaaaagg gcgatcgccg tgtcttgggg tagttatagc aagctcccaa
     5881 tagaaggatt catgctgata agcggcacga cgaccttggc gttttttagt ctgttcgtaa
     5941 ccagtccggc gttgtacttc tttggtaaat gctttatagc tcggagcaat tacaccagct
     6001 tcttcgcaag ctcgaactaa agccgcgtga acctcccaca ttcttttttg tttaagagtt
     6061 tcatagtctt ctaaaataaa cttctccata attgctaagg tgtcttcggg cagcttgcgc
     6121 tgtcgattac ctttgacact gcgatgggac aaaagcccaa tgtagccgca accgtatttt
     6181 tgaattgcaa ttaaatattt agctttccag tgacgaatgg tacgggctgg tattgtttcg
     6241 ttttctggag gatgtccgtt taagtaaggt tcgatagttt tgtagcgttc aagggcttct
     6301 gcttggtctt caggactagc ttgatgaaat cgctgccaag attctgtgct aatagctgtt
     6361 ttttctggtg tttggagact tgtgattttt ccttggcgaa ccagattctc aaatatagct
     6421 tttttgagtt caattaactg attattttct gcgccaagta taacctctgt ttctccaaca
     6481 tggatgatat ttaaaccttt gccgtcccaa ctaacagttg ctccagtaac aatattgact
     6541 acaggagaga tagccggaag acttacactt ggttgctcaa ccattaatct ataagcacta
     6601 gcagtttgct cgtcacggaa gattagacat ctttctggtt cagctagtgg tgtagcattt
     6661 aaatcaatat agatttgctc ttgaataatt aaggtgtaga tatcgtcaga tttagctcct
     6721 gttgaccgat tgagtaattc tgctaaagtg attcctggtt gagatgacac aattgcaagt
     6781 aatgattgag cgatcgcttc ttccactact agagattctg agcgataata atcttctaaa
     6841 aattgcaaat tacgttgcag aacccaattg atttcgcgat ctgaatgaat gcggtgataa
     6901 agcccaaact ggttagcata aacttctcct ggcaaggaat gccattgatt attttcttca
     6961 gagtaaaagt aacgattggg gtttttctct gctagttttt tcaaatcctg ctcagtttta
     7021 cactcttccc aaccagccga attggttcgg atgacaaata aatctggggt gtgtaaaacg
     7081 cctaatctac gtccgttact tgcttgatag tcaagtttaa tttgcggtgg ttgatcgtag
     7141 aattctagaa catctttatc atgttctaac tgatagataa ttggcagttc aacccgatga
     7201 gactcaaatt gaatcgtgac tcccatcttg cgactagggt agcgtcccga aacattcttg
     7261 ctaccgccac caacgcgacg ggatggctca gatgagcgaa ctttctcaat aattttttgc
     7321 cctgcctcgg atatttgaag gcgagcaacc cattgttcaa attgctcttt catggtatat
     7381 ctccttatta aagttaaaca aaattatttc tacaggggaa ttgttatccg ctcacaattc
     7441 ccctatagtg agtcgtatta atttcctaat gcaggagtcg cataagggag agcgtcgaga
     7501 tcccgagggt cgttaaatag ccgcttgaca ccatcgaatg gcgcaaaacc tttcgcggta
     7561 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     7621 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     7681 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     7741 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     7801 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     7861 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     7921 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     7981 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     8041 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     8101 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     8161 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     8221 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     8281 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     8341 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     8401 cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     8461 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     8521 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     8581 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     8641 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8701 aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc
     8761 aacccagtca gctccttccg gtgggcgcgg ggcatgacta acatgagaat tacaacttat
     8821 atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga
     8881 gcggttcagt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     8941 tttgccctgt aaac
//
