LOCUS       sequence-330379-        7672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of AvCAST Cascade proteins. Two BsaI sites for 
            spacer cloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7672)
  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 7672)
  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: pMS7: pCascade(AvCAST)_entry
FEATURES             Location/Qualifiers
     source          1..7672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(215..1297)
                     /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     1259..1278
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1298..1375)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1421..1439
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1440..1464
                     /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        1703..1721
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1722..1746
                     /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(5711..5729)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5725..5772
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5943..6734)
                     /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(6735..6826)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgatcttttc tactgaaccg ctctagattt cagtgcaatt tatctcttca aatgtagcac
       61 ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca
      121 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
      181 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      241 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      301 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
      361 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
      421 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
      481 atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
      541 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
      601 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
      661 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
      721 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
      781 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
      841 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
      901 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
      961 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     1021 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     1081 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     1141 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     1201 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     1261 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     1321 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     1381 tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact cactataggg
     1441 gaattgtgag cggataacaa ttcccctgta gaaataattt tgtttaactt taataaggag
     1501 atataccatg gtgctttaac attagatgtc gttaggcgtt gagcagagac ctctggtctc
     1561 gtgctttaac attagatgtc gttaggcgtt gagcagatcc gaattcgagc tcggcgcgcc
     1621 tgcaggtcga caagcttgcg gccgcataat gcttaagtcg aacagaaagt aatcgtattg
     1681 tacacggccg cataatcgaa attaatacga ctcactatag gggaattgtg agcggataac
     1741 aattccccat cttagtatat tagttaagta taagaaggag atatacatat gattattgat
     1801 agtaacttag ttacgcagct tccctcagaa tcatcttcaa agccaagaga gcttttaatg
     1861 tctcagtttc catatatgga tttaactttt agccttacgg gtgagacatt acctttcgac
     1921 cacggctatg aattattttc tgcgatcgcc cactttgaac ccaaactaca tacattagac
     1981 acgttgggaa ttcagaccat tgcaggtatt cccaaggatg gtgtaattaa tctgacccaa
     2041 aattctaggc ttcgtattcg gataccagta aatcaggtac gtttggttta tccattggcg
     2101 ggtaagtcac tgagaattgg caagcataca attcgtttgg gtattcctga catctgtttg
     2161 cttcaacctg cggagaaact gcgatcgcgc attgtggtaa taagaggcta tgaagaacct
     2221 gaaagttttt tagtcgtagc gcagcgtcag ctagaacaac tgggtatcca agcaattgct
     2281 agcattccca ccaaagccaa tggcaaacca atacgtagga ctatcaaaat taagcgattt
     2341 acagtcgtgg gttttggatt agaagttata aatttaagcg atgaagactc actgaggttg
     2401 caaatacatg gtgtaggagg aaagcaaaag atgggttgtg gactcttcat gcctattaag
     2461 gaaaggcaat agcaattact cgaaagcaga tgatgaagag agcagataac cccaaaattc
     2521 agcttaatct cggcgatcct agtatgactt tattgcatcg tgctggggtt gcaggacttt
     2581 ggatgacgct caaaaaactg gaaaaaatct atccaacacc tgctgaacgt attggtaatt
     2641 taacttggtt gctaacccct cactatatca atcttgattg ggaaggacag gatttcacag
     2701 ttttagactg gttgctgaag caatcatttc aaatcagcga tgagggttta atttccctga
     2761 cagggttaga ttctcaaaac atggatattc aaaatcaaat aattatccat caaggtatca
     2821 cagccacttt tacacaacat aataaatttt ttaagtctgc gggtcaacaa tctaagcaac
     2881 taattattga tggtattcag gttagggttg attataagaa agctgcttcc tatgctcatc
     2941 aacattttgc taaacattta tgtgaccagt acggacaatt gctaaaagag ccaattggta
     3001 tcaggggatg gctctatcca ggagcagtag ttcgccatta tgcttttaaa gaacaaacag
     3061 catttgaaga aaaagcagaa tatgctctag cgttgttgtt cgctcctatg gcttgtcaat
     3121 attttgtgct gcgatcgcac accgaaccat cacatacaaa ctatgttttg gttattcctg
     3181 aagtaattga tttagaactt tatgcacatt ttttgtggtg tttgggtaac ttagattata
     3241 aacattttca cgtatctagc ttaggagatg ctggattaaa atttcttaat tataaaacaa
     3301 cacaaatgcc tacttccaat tttttaaaac gatgtcaaat aatctttttt agtacaaaag
     3361 cttggtcaga acagcaaaaa actcgtgcag aaatagcaat catagaatta aattcttcag
     3421 ttaattatct ttataagctt agttgtattt gttttccaga atgtcaaatt gttttatata
     3481 aaaatcaaca ttatttgctt ccaagtttaa taaaaggaat aatcgccaat aatctagcaa
     3541 tgggtcggtc ttggtgggct aattggattt catttattaa aaataaaaaa tcatttaaac
     3601 aatttactga tgagtataaa ggaattaata agatgattca ggactcagaa tgggatatag
     3661 aagctcaaaa gctatttatt caagcttgtc atgaagcatt aagaaaaata tatgccaaaa
     3721 tttatagcag aacaaaagaa ggtgaatacg ctcaaataga acgagaaaat acacgcattt
     3781 tatctcagtt aaaacgctgc accaatactg acaatttccg aaaatttatt gcagagtttt
     3841 ggggaagggc agggcaaata gcaattctgg aagaacattg gatggaatta cttccattga
     3901 ctacagggat aggcaattgg aaagtggcca gggacttaac ttttattgca atggctagct
     3961 acccaaagaa taaaataaca gaggtagagg taaaaccatc acaaatatcg gagctaaata
     4021 ctgaataaac ttcatatttt ttgatttgat taagaacaaa atattaattg tttaggtaat
     4081 ggtggaatgt ttcatctgtt tggtaatatt ttgactagtt atggaaccgc cgctaataat
     4141 cgtggtgaaa atgaaggtaa taccactacg ttacaaaaat taatttggaa aggtgaagtt
     4201 cactctactg tttcatctga ggcaattcgt tgggcattgc gatattattg gcaaaatgct
     4261 ggctatcctg tgaatagacg ttgggatgaa aatgctcaac cagtacccga tcacatcttg
     4321 caagatccta attttgatga tattcgcttc attgatgatg atgtcttggg atttatgcaa
     4381 gcggaagcag caaaagttga agcagcagtt gaatccgaag ttgaaatcca gaactcaact
     4441 caagatgagg acaaaaaact ttctcaaggt aaggagaaag aaactgcaaa acgtaagcgt
     4501 aaggaaaaac ctaaagggaa aataactact agacgaggtg tattagaagt cactcgtgca
     4561 gtttcaacaa ttccttatgc tggtgatctt actttcaatg ctatgagtgg aaagaaagga
     4621 cgaacatctc tttattctac agaagttcat gctactcgat atcaatatgg cttcgcatta
     4681 acacccaata gcctcaaaga taaatctcgc attaatgctg tcctcgatag cttaatttct
     4741 attggagaag tggcaggaaa tcacgctcgt tttctctatg atttttctcc tgatagcatc
     4801 atattacggt ggacacacga tttctcacct cggttacttt actgttttga agaagatgaa
     4861 cttagaaata tatcaacacc ggatttagta aggcgggtag aatctggaga tatagaccct
     4921 aaagaattat ggataggagg ggcaatttct agcactctgg aagatttagg ggcaaatgtg
     4981 tttcccggag tgaagacaac agttgaagca ttaaaacagg tgattattga agatttgcag
     5041 ttgtcgttaa ggagttcatg atatgacgac aattgcctta aaagtagaag ttcccatagc
     5101 ttgttttcgg cagtctcgtg ctagggaata tggcgaaact taccctgtac caccaccatc
     5161 gacagtatat ggaatgttac tatctcttgt tggagaagtg gatagataca aacactgcgg
     5221 tgtgaagctg gcgatcgcac ttctatctga accagaaaag tcaactgtta ttcgcacctt
     5281 tcaccgattt aagacgaaga atatccatga ctctaaaaat aataaacctg attatcagga
     5341 gttattaact aatatagaat ttatagtttg ggtggatgct ggagtagata aagctaaacc
     5401 aaatttagtg gaacgtttag cagaggcatt gactaacccg gcttctatta ataggtttgg
     5461 tggattatgt ttaggtgaaa gtcgggattt agtgaatgat gttacccttt tacctaaaaa
     5521 ttactatgca gaatcaatgc gatggttaat tcgagatgaa tatgggttac taactttacc
     5581 ttactgggta gaccatgtag gttcgcgggg aacgcgatgg ctacgttacg agattcagaa
     5641 atgtccagtc tttcaaccac ctgaattatc ttggacttca gttcaaagta attagcctag
     5701 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     5761 aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact gcttccggta
     5821 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
     5881 ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg aattgttaga
     5941 cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa ttcttccaac
     6001 tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg tctagcttca
     6061 agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc agcgacatcc
     6121 ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt aagcactaca
     6181 tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt ttcatttagc
     6241 gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc tggacctacc
     6301 aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg
     6361 gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg
     6421 cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt gacttctaca
     6481 gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa
     6541 gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc aatatcactg
     6601 tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag caacgtcggt
     6661 tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac ttcggcgatc
     6721 accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6781 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc tagctcactc
     6841 ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc ggacacatac aaagttaccc
     6901 acagattccg tggataagca ggggactaac atgtgaggca aaacagcagg gccgcgccgg
     6961 tggcgttttt ccataggctc cgccctcctg ccagagttca cataaacaga cgcttttccg
     7021 gtgcatctgt gggagccgtg aggctcaacc atgaatctga cagtacgggc gaaacccgac
     7081 aggacttaaa gatccccacc gtttccggcg ggtcgctccc tcttgcgctc tcctgttccg
     7141 accctgccgt ttaccggata cctgttccgc ctttctccct tacgggaagt gtggcgcttt
     7201 ctcatagctc acacactggt atctcggctc ggtgtaggtc gttcgctcca agctgggctg
     7261 taagcaagaa ctccccgttc agcccgactg ctgcgcctta tccggtaact gttcacttga
     7321 gtccaacccg gaaaagcacg gtaaaacgcc actggcagca gccattggta actgggagtt
     7381 cgcagaggat ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc aaagtccggc
     7441 tacactggaa ggacagattt ggttgctgtg ctctgcgaaa gccagttacc acggttaagc
     7501 agttccccaa ctgacttaac cttcgatcaa accacctccc caggtggttt tttcgaaggt
     7561 taagtcagtt ggggaactgc ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
     7621 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tt
//
