LOCUS       sequence-330689-        9185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of AvCAST TnsA, TnsB, TnsC and TniQ proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9185)
  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 9185)
  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: pMS10: pTns(AvCAST)_Delta-TnsD
FEATURES             Location/Qualifiers
     source          1..9185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      916..1551
                     /direction=RIGHT
                     /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     1387..1406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1569..1660
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1661..2476
                     /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             1661..2476
                     /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     complement(1734..1753)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2706..2753)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2749..2767
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    2831..2855
                     /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(2856..2874)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8654..8678
                     /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(8679..8697)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        8743..8820
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(8821..9185,1..718)
                     /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(8840..8859)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
       61 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
      121 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
      181 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
      241 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
      301 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
      361 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
      421 gcgcgttggt gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta
      481 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
      541 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
      601 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
      661 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
      721 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
      781 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact aacatgagaa
      841 ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga taaattgcac
      901 tgaaatctag agtgatggtg tcgggaatcc gtaaaggatc ttcttgagat ccttttacga
      961 tcgtcgtaat ctcctgctct gtaaacgaaa aaaccgcctg gggaggcggt ttgatcgaag
     1021 gttaagtcag ttggggaact gcttaacctg gtaactggct ttagtggagc gcagatacca
     1081 aatactgtcc tttcagtgta gcctctgtta ggccaccact tcaagactct cgatatctaa
     1141 atccactaat tctcagttac caatggctgc tgccagtggc gttttgtcgt gtctttccgg
     1201 gttggactca agatgatagt taccggataa ggcgcagcag tcgggctgaa cggggggttc
     1261 ttgcacacag cccagcttgg agcgaactgt ctacacggaa cgggacgtgg tgatttgggt
     1321 aaagcctcca ccacaacacg gacgccgcag gacgggaaca ggagagcgca agagggagcc
     1381 atcaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga
     1441 gcgtcagatt tcgtgatgtt cgtcaggggg gcggagccta tggaaaaacg gcttcgctcc
     1501 ggccttattg tctctctgct aagtatcctc ctggcatctt ctaggacgtt tctgcgctag
     1561 catgcctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1621 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagccata ttcaacggga
     1681 aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     1741 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     1801 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     1861 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     1921 tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt
     1981 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2041 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg accgcgtatt
     2101 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2161 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2221 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2281 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2341 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2401 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2461 cgatgagttt ttctaagaat taattcatga gcggatacat atttgaatgt atttagaaaa
     2521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacttgcg gagacccggt
     2581 cgtcagcttg tcgtcggttc agggcagggt cgttaaatag ccgcttatgt ctattgctgg
     2641 tttaccggtt tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggt
     2701 ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc
     2761 tcagcggtgg cagcagccta ggttaattaa gtatatctcc ttcttatact taactaatat
     2821 actaagatgg ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg
     2881 attatgcggc cgtgtacaat acgattactt tctgttcgac ttaagttatg actgaaagaa
     2941 ctcttctata gaatttgcta aaaattttat cattaagata tgagttatgc ggtcaattgc
     3001 tgggtttaag tcgcatccaa acaagcagtg agaaaaatat ttaccttgat tttgggtcag
     3061 ctttggattg agggcttcta aggattcttg accataaaac tcaaatataa gctgggcaaa
     3121 accacaatca ttaaacttaa acttgccacc agcaaaggtt tttagcagtc ctttatccaa
     3181 taagtaacgc tggtactgag cgcgaagcca agccaaacct tgaaaggcaa aactgctatg
     3241 aattagccaa ttaatatcat gtgccagtat tgaaagtgtt tctacagttt tatttgtcaa
     3301 ttttttcaca ttatttgcta ataaacaatt atctatgctg gcagcatgat aatgtatgcc
     3361 tttcgattca acttctatac agctattgtg taaaacaaca cgatgaattg gacaaactaa
     3421 aatgccaggg acttgatgta ttctatgcca gtaaggttca ccatactttt gggtatcttt
     3481 ctctaaacat tcagggcaaa attttaaaaa tttttttgaa tcatcactag aattattagc
     3541 cactttagca aaatctagta tagacccact gaatttatta ctcattgagt ttttcagcag
     3601 ccaagcctcc tgcggcatta ataaagttgc ataaaaggaa tataaagtat gattttgtag
     3661 taaattctca actttttgct gggagcgtaa aggtaagttt ttcactaaat aatttaaatt
     3721 attagttaag gtaattttgc atatttgttg ggaactgtaa ccgaatagct ctaagtcagt
     3781 ttgcttgaaa cttttatttc cactccgaat atggtatcta gctaaagtgc tatataaaag
     3841 ttcatcggga tacagagttg ggaaaaaact aagcatttag aagctaaata ttgataaaat
     3901 cttcaactgg atttttgatt atacctgcta atttcaaatc ctcgtatgca gattttttgt
     3961 ctttcttggc attctcgaca attcgccgaa tatcattctc ttggtacttt gtctttaatt
     4021 tagctttcct ggtacttgtt ttagcaagac tctcatttac ttgacatcct tgcaaagcaa
     4081 gagcatatgc cttattaact aaagatggaa ttggaatgtc tggatcttca tgactataaa
     4141 caatttgttc tgcacattcc tttgctttac cagattctac ttccaatttc aacagctcac
     4201 taattacata attgagcttc ggagatactt tctgctgatt ctgctgcttt ttagctaatc
     4261 tcttaatctc tgtcaaatct ttagcatcaa tctcagatac acatctattg tgatattccg
     4321 taatatcgag aggagcaata tccttatatt taatcatcca ctctctgtcc cctaacctaa
     4381 ttgcatctag cattggcttt actaaataca atccttcttt agctacttga cgaatcagat
     4441 caaccgtaat aatttcttgg tcacctattg aaatcgctct ccattgaacc attttgtaaa
     4501 gttttatagc aatatcaata attccctggc tttcatcata aaaaacttct tttatttcat
     4561 ccgagaaatg aacaggtgtt tttgtccatt ggtattccca tattccttct aaaaagtaac
     4621 accattcatc attatttacc attctatccc acattctact tccttctcca gtatttcttc
     4681 tagcatttct aagattacct tgaagaaaac taatggcttc attcgtacct acacgaataa
     4741 ctggaattcc aatagtatta tcgagcttta ctaaaaaatt taatatttcg tctctatttc
     4801 tttttgcagt tgctaaattt tgtatttcat caatcaccaa aactcctaga tgatgagtgt
     4861 gtgctgtttg tgcaagttga gccaatagat aatcttcaga tgtgcgtgct gggttaaatt
     4921 tctttaataa attagttcct aacaatttat cgatacctgc aaaagtatct atacataacc
     4981 cctttaaagt tttagggcaa tctacccgca gccaaactat ttggtaaaca ctataatcag
     5041 gatgaaaaat tatttgggga taaagctcaa gaatattttt gaaatttgta gtctttccaa
     5101 tacctgatgg gccaattaga gttaagccgg acgcagatgt aggaagatta acatagtttt
     5161 ctaaattttt acctcctcca ttacctattg cattataaat ttgcctagag cgatttgcat
     5221 attcaggttt taaaaggttc cttgacagat agcctgtcat gataagcgta caaataactt
     5281 tttgtaattc aagagtttta ttttgagggt caaaatatct agataacctc tcaatacaat
     5341 gaaatctata atgtggttct agctctctct cttgttcatc aaaatttggt tctctggcta
     5401 ctttatctat aaattctgct gctgatagga taggaggtaa tgcctgaatt aatggattag
     5461 tgttgtattc tggcagtttt tgatctatat atctggcaat aactgctagt tctccattag
     5521 gaattttaac cagttcactc tcaattctgt ccacgttttc gctcctgtcg ctttttctta
     5581 agaagattca gatggtttgg ctgtagtgat tctggctgtt cggaggtttc tgctgtactc
     5641 tctgcaacct gaaaactgct ctgattatct gtaattgcta attcaaatcc ttcattctct
     5701 cgtcgtttag ccttttcaaa agccctatta tctcttattc cagaaacttt cttgctctta
     5761 cttaaattat gatattgaga agcatcagtc attttttctg ctctatctac tatgctttca
     5821 atatcagcta ttaaatctac cttttcttgt agctgtttac cttggctttt ctgtctttgc
     5881 aattcttcat aagcaagcag atattcaata tcatgaaagt ttttattagc ataccgccct
     5941 tctaagtcaa taagaaaaca tttttcaaaa tctctaccat cactagatgg aagataaatg
     6001 aagtttggct ttctgatgtc ataagataca atcaatgatt tttctgactt agaaaggaga
     6061 ttacttctag ctttttcaaa ccattgctca tttttagcac tatcacaggt atagtacatc
     6121 tccttacctc tcattttgat gcctttagca gtaatagttg ctttctcagt aggcatcaag
     6181 ttcaatttga caatatcttc agaaaaagtt ctgagtctgc cagaacgatt ggcgataccc
     6241 cacttccaca actctctagg tataggaggg gtatcatcag caatcattgc ttcatctctt
     6301 tcatattcag taagaggttt attgttatgg tgaacaatta aacgaataat aatctcagta
     6361 aattgatcta tatcaagttt gccatcaagc ctataatccc ttgcacctct ttgcctaaaa
     6421 tccgtatcca cataaccagg taaaaaaggt tttacatatt catgaattat acgaaagtgt
     6481 ctttctacta accctttcca atctgctcta taggaagctg cattttcaat acgaacataa
     6541 agattaggga ttaatgtttc tactgtcata cctgctaatt ctcccctatc tcctaaaatt
     6601 gcatctggca tttgattgca aggccattca ttttcagtaa tattaatgtt atattcctgg
     6661 caaaacaggg ttttatctgt agctacattg gctagtgcca tcattgctcc tatccaagat
     6721 gggccttcta aaccgacata aacacctgta atcattctac taaaaacatc aataattaca
     6781 tagataactg gtctaccaat tatccagtca cggttatatt ttgaaactaa atatacatca
     6841 gcaactgtgg catcaatttg atatctagaa ccagggccaa tagtttccat cttagagttt
     6901 cccaaaattg ctctattctc taaagctact cgcttggctc cttttcgaga tgttaaagtt
     6961 ttggtaacat catcctgttc aagttgatac cagtatttga attgagtaat tgtaggtatt
     7021 ttatcagttg gggttaatat agatttttga acaccattct cgtcatatct aattccttcc
     7081 ttgtaatatt ctttcaccat taattcatag gctgttgtta agtagttttc ttttggatta
     7141 ttgtaaaatt tgcttataga tactctaaaa atctttctat cttcttctgt aacatttaca
     7201 ccttcaccaa tatctgggtc atgtgcatat tttcttggtc ttcctctctt cttgcccgat
     7261 tgtcctcgaa tcttgccctt acctgatgaa ttgataaaat caggtatgag agcatttata
     7321 gtttttcctc tctgccaaaa tcttctcaaa taaccataga cggttttttc tactaattta
     7381 tctttatccc tttcttcgtt gtacttttca accgcttctt taaccatcga tcctctaaaa
     7441 tttctataat atattgatgg ttcttctata attaatgatg aaattatctc ccaagcttta
     7501 tctctgtatt ccttttcctt atctgtcaaa ttttcatccc gaataattct tgcccaagga
     7561 tcttgttgct gcttaaaagc aataccttca gttatagctt ctaaaatatc tgatattttc
     7621 cttggttcag gtagaccttt cttagcatta atatctataa caaatgcaat aatgtaacct
     7681 tcatctatcc aaataattct ttcgactata tgagtgtttt ggtatacaat ccactctatt
     7741 aaatcattga caaataaatc tttaaccata ttatttacct atcatgtggg tctaaaatat
     7801 tattttttca ataactttag ttgacggaca agcagataat tttattgcta ctagatccac
     7861 aaggatttct ttcctagcaa ttaaatgttt aaatacataa agtgatgtac cggaatttat
     7921 attcatttct ttgtctaaag cagtagttac tctatttata gttgtatctt cttcctgaag
     7981 cttagatttt aaaattgttg acaattcact cagttgtgca gtgtccattt cagaatttgc
     8041 ttctagccta taagcagaat gaacccactc aacattttca gctaaaagcc tgggaatttc
     8101 tttttctgtg ataattcccc agtcaatacc ctttgctaag taatattgtc tttctatctc
     8161 gaatttctca ttaacttgct ttttctccag ttctgttgac ggcttaacag tccgggcaat
     8221 ctgcatctgc ttcccatcgc ggttgactgt aagcataaag tcgcttgtta aaacgtgagg
     8281 agtgccactt tgtgtatcta cagggtactt tatacctatg tctgtagcaa tactcatggc
     8341 tagatcgaga ttgtctaaag gaaattgctc tcttatatct gtgactatat ctgaccactc
     8401 aaacaaataa aatagccgtt tttcatggtc tgagaagaaa tgatgtactc gatttgtttt
     8461 ccaaccagga gttctagaag aacgtccttt agagggaaca tcttggattg taagccaagg
     8521 tttatagtgc ttaccacttc ctcggccacg accttctttt atatatcgct catatttagc
     8581 ttgagtccat tcccttctac ccctaaccat ggtatatctc cttattaaag ttaaacaaaa
     8641 ttatttctac aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt
     8701 tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc
     8761 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat
     8821 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt
     8881 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg
     8941 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag
     9001 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc
     9061 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa
     9121 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt
     9181 gggct
//
