LOCUS       sequence-330377-        8234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of AvCAST TnsA, TnsB and TnsC proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8234)
  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 8234)
  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: pMS11: pTns(AvCAST)_Delta-TniQ,Delta-TnsD
FEATURES             Location/Qualifiers
     source          1..8234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     563..582
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(602..679)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        725..743
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    744..768
                     /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        5597..5615
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5616..5640
                     /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(5704..5722)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5718..5765
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5995..6810)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5995..6810)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6718..6737
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(6811..6902)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6920..7555)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(7065..7084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7753..8234,1..601))
                     /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"
ORIGIN
        1 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
       61 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
      121 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
      181 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
      241 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
      301 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
      361 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
      421 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
      481 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
      541 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
      601 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
      661 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      721 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tgtagaaata
      781 attttgttta actttaataa ggagatatac catggttagg ggtagaaggg aatggactca
      841 agctaaatat gagcgatata taaaagaagg tcgtggccga ggaagtggta agcactataa
      901 accttggctt acaatccaag atgttccctc taaaggacgt tcttctagaa ctcctggttg
      961 gaaaacaaat cgagtacatc atttcttctc agaccatgaa aaacggctat tttatttgtt
     1021 tgagtggtca gatatagtca cagatataag agagcaattt cctttagaca atctcgatct
     1081 agccatgagt attgctacag acataggtat aaagtaccct gtagatacac aaagtggcac
     1141 tcctcacgtt ttaacaagcg actttatgct tacagtcaac cgcgatggga agcagatgca
     1201 gattgcccgg actgttaagc cgtcaacaga actggagaaa aagcaagtta atgagaaatt
     1261 cgagatagaa agacaatatt acttagcaaa gggtattgac tggggaatta tcacagaaaa
     1321 agaaattccc aggcttttag ctgaaaatgt tgagtgggtt cattctgctt ataggctaga
     1381 agcaaattct gaaatggaca ctgcacaact gagtgaattg tcaacaattt taaaatctaa
     1441 gcttcaggaa gaagatacaa ctataaatag agtaactact gctttagaca aagaaatgaa
     1501 tataaattcc ggtacatcac tttatgtatt taaacattta attgctagga aagaaatcct
     1561 tgtggatcta gtagcaataa aattatctgc ttgtccgtca actaaagtta ttgaaaaaat
     1621 aatattttag acccacatga taggtaaata atatggttaa agatttattt gtcaatgatt
     1681 taatagagtg gattgtatac caaaacactc atatagtcga aagaattatt tggatagatg
     1741 aaggttacat tattgcattt gttatagata ttaatgctaa gaaaggtcta cctgaaccaa
     1801 ggaaaatatc agatatttta gaagctataa ctgaaggtat tgcttttaag cagcaacaag
     1861 atccttgggc aagaattatt cgggatgaaa atttgacaga taaggaaaag gaatacagag
     1921 ataaagcttg ggagataatt tcatcattaa ttatagaaga accatcaata tattatagaa
     1981 attttagagg atcgatggtt aaagaagcgg ttgaaaagta caacgaagaa agggataaag
     2041 ataaattagt agaaaaaacc gtctatggtt atttgagaag attttggcag agaggaaaaa
     2101 ctataaatgc tctcatacct gattttatca attcatcagg taagggcaag attcgaggac
     2161 aatcgggcaa gaagagagga agaccaagaa aatatgcaca tgacccagat attggtgaag
     2221 gtgtaaatgt tacagaagaa gatagaaaga tttttagagt atctataagc aaattttaca
     2281 ataatccaaa agaaaactac ttaacaacag cctatgaatt aatggtgaaa gaatattaca
     2341 aggaaggaat tagatatgac gagaatggtg ttcaaaaatc tatattaacc ccaactgata
     2401 aaatacctac aattactcaa ttcaaatact ggtatcaact tgaacaggat gatgttacca
     2461 aaactttaac atctcgaaaa ggagccaagc gagtagcttt agagaataga gcaattttgg
     2521 gaaactctaa gatggaaact attggccctg gttctagata tcaaattgat gccacagttg
     2581 ctgatgtata tttagtttca aaatataacc gtgactggat aattggtaga ccagttatct
     2641 atgtaattat tgatgttttt agtagaatga ttacaggtgt ttatgtcggt ttagaaggcc
     2701 catcttggat aggagcaatg atggcactag ccaatgtagc tacagataaa accctgtttt
     2761 gccaggaata taacattaat attactgaaa atgaatggcc ttgcaatcaa atgccagatg
     2821 caattttagg agatagggga gaattagcag gtatgacagt agaaacatta atccctaatc
     2881 tttatgttcg tattgaaaat gcagcttcct atagagcaga ttggaaaggg ttagtagaaa
     2941 gacactttcg tataattcat gaatatgtaa aacctttttt acctggttat gtggatacgg
     3001 attttaggca aagaggtgca agggattata ggcttgatgg caaacttgat atagatcaat
     3061 ttactgagat tattattcgt ttaattgttc accataacaa taaacctctt actgaatatg
     3121 aaagagatga agcaatgatt gctgatgata cccctcctat acctagagag ttgtggaagt
     3181 ggggtatcgc caatcgttct ggcagactca gaactttttc tgaagatatt gtcaaattga
     3241 acttgatgcc tactgagaaa gcaactatta ctgctaaagg catcaaaatg agaggtaagg
     3301 agatgtacta tacctgtgat agtgctaaaa atgagcaatg gtttgaaaaa gctagaagta
     3361 atctcctttc taagtcagaa aaatcattga ttgtatctta tgacatcaga aagccaaact
     3421 tcatttatct tccatctagt gatggtagag attttgaaaa atgttttctt attgacttag
     3481 aagggcggta tgctaataaa aactttcatg atattgaata tctgcttgct tatgaagaat
     3541 tgcaaagaca gaaaagccaa ggtaaacagc tacaagaaaa ggtagattta atagctgata
     3601 ttgaaagcat agtagataga gcagaaaaaa tgactgatgc ttctcaatat cataatttaa
     3661 gtaagagcaa gaaagtttct ggaataagag ataatagggc ttttgaaaag gctaaacgac
     3721 gagagaatga aggatttgaa ttagcaatta cagataatca gagcagtttt caggttgcag
     3781 agagtacagc agaaacctcc gaacagccag aatcactaca gccaaaccat ctgaatcttc
     3841 ttaagaaaaa gcgacaggag cgaaaacgtg gacagaattg agagtgaact ggttaaaatt
     3901 cctaatggag aactagcagt tattgccaga tatatagatc aaaaactgcc agaatacaac
     3961 actaatccat taattcaggc attacctcct atcctatcag cagcagaatt tatagataaa
     4021 gtagccagag aaccaaattt tgatgaacaa gagagagagc tagaaccaca ttatagattt
     4081 cattgtattg agaggttatc tagatatttt gaccctcaaa ataaaactct tgaattacaa
     4141 aaagttattt gtacgcttat catgacaggc tatctgtcaa ggaacctttt aaaacctgaa
     4201 tatgcaaatc gctctaggca aatttataat gcaataggta atggaggagg taaaaattta
     4261 gaaaactatg ttaatcttcc tacatctgcg tccggcttaa ctctaattgg cccatcaggt
     4321 attggaaaga ctacaaattt caaaaatatt cttgagcttt atccccaaat aatttttcat
     4381 cctgattata gtgtttacca aatagtttgg ctgcgggtag attgccctaa aactttaaag
     4441 gggttatgta tagatacttt tgcaggtatc gataaattgt taggaactaa tttattaaag
     4501 aaatttaacc cagcacgcac atctgaagat tatctattgg ctcaacttgc acaaacagca
     4561 cacactcatc atctaggagt tttggtgatt gatgaaatac aaaatttagc aactgcaaaa
     4621 agaaatagag acgaaatatt aaatttttta gtaaagctcg ataatactat tggaattcca
     4681 gttattcgtg taggtacgaa tgaagccatt agttttcttc aaggtaatct tagaaatgct
     4741 agaagaaata ctggagaagg aagtagaatg tgggatagaa tggtaaataa tgatgaatgg
     4801 tgttactttt tagaaggaat atgggaatac caatggacaa aaacacctgt tcatttctcg
     4861 gatgaaataa aagaagtttt ttatgatgaa agccagggaa ttattgatat tgctataaaa
     4921 ctttacaaaa tggttcaatg gagagcgatt tcaataggtg accaagaaat tattacggtt
     4981 gatctgattc gtcaagtagc taaagaagga ttgtatttag taaagccaat gctagatgca
     5041 attaggttag gggacagaga gtggatgatt aaatataagg atattgctcc tctcgatatt
     5101 acggaatatc acaatagatg tgtatctgag attgatgcta aagatttgac agagattaag
     5161 agattagcta aaaagcagca gaatcagcag aaagtatctc cgaagctcaa ttatgtaatt
     5221 agtgagctgt tgaaattgga agtagaatct ggtaaagcaa aggaatgtgc agaacaaatt
     5281 gtttatagtc atgaagatcc agacattcca attccatctt tagttaataa ggcatatgct
     5341 cttgctttgc aaggatgtca agtaaatgag agtcttgcta aaacaagtac caggaaagct
     5401 aaattaaaga caaagtacca agagaatgat attcggcgaa ttgtcgagaa tgccaagaaa
     5461 gacaaaaaat ctgcatacga ggatttgaaa ttagcaggta taatcaaaaa tccagttgaa
     5521 gattttatca atatttagct tctaacttaa gtcgaacaga aagtaatcgt attgtacacg
     5581 gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     5641 ccatcttagt atattagtta agtataagaa ggagatatac ttaattaacc taggctgctg
     5701 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     5761 ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     5821 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     5881 caagctgacg accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct
     5941 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
     6001 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     6061 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     6121 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     6181 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     6241 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     6301 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     6361 agcgagacga aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     6421 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     6481 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     6541 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     6601 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     6661 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     6721 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
     6781 gcaagacgtt tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat
     6841 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6901 aataggcatg ctagcgcaga aacgtcctag aagatgccag gaggatactt agcagagaga
     6961 caataaggcc ggagcgaagc cgtttttcca taggctccgc ccccctgacg aacatcacga
     7021 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7081 tccccctgat ggctccctct tgcgctctcc tgttcccgtc ctgcggcgtc cgtgttgtgg
     7141 tggaggcttt acccaaatca ccacgtcccg ttccgtgtag acagttcgct ccaagctggg
     7201 ctgtgtgcaa gaaccccccg ttcagcccga ctgctgcgcc ttatccggta actatcatct
     7261 tgagtccaac ccggaaagac acgacaaaac gccactggca gcagccattg gtaactgaga
     7321 attagtggat ttagatatcg agagtcttga agtggtggcc taacagaggc tacactgaaa
     7381 ggacagtatt tggtatctgc gctccactaa agccagttac caggttaagc agttccccaa
     7441 ctgacttaac cttcgatcaa accgcctccc caggcggttt tttcgtttac agagcaggag
     7501 attacgacga tcgtaaaagg atctcaagaa gatcctttac ggattcccga caccatcact
     7561 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
     7621 taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     7681 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     7741 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7801 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     7861 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     7921 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     7981 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc
     8041 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     8101 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     8161 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     8221 gagatattta tgcc
//
