LOCUS       sequence-330861-        7576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of PmcCAST TnsAB and TnsC proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7576)
  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 7576)
  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: pMS28: pTns(PmcCAST)_Delta-TniQ,Delta-TnsD
FEATURES             Location/Qualifiers
     source          1..7576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(600..1682)
                     /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     1644..1663
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1683..1760)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1827..1845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1846..1870
                     /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        5816..5834
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5835..5859
                     /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)."
     CDS             5968..6012
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(6050..6068)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6064..6111
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6232..7023)
                     /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(7024..7115)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgctgcgcc ttatccggta actgttcact tgagtccaac ccggaaaagc acggtaaaac
       61 gccactggca gcagccattg gtaactggga gttcgcagag gatttgttta gctaaacacg
      121 cggttgctct tgaagtgtgc gccaaagtcc ggctacactg gaaggacaga tttggttgct
      181 gtgctctgcg aaagccagtt accacggtta agcagttccc caactgactt aaccttcgat
      241 caaaccacct ccccaggtgg ttttttcgaa ggttaagtca gttggggaac tgcttaacct
      301 tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga
      361 aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta gatttcagtg
      421 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
      481 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
      541 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
      601 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      661 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
      721 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
      781 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
      841 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
      901 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
      961 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     1021 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     1081 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     1141 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     1201 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     1261 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     1321 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     1381 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     1441 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     1501 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     1561 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     1621 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     1681 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     1741 ttgcgccatt cgatggtgtc aagcggctat ttaacgaccc tcgggatctc gacgctctcc
     1801 cttatgcgac tcctgcatta ggaaattaat acgactcact ataggggaat tgtgagcgga
     1861 taacaattcc cctgtagaaa taattttgtt taactttaat aaggagatat accatgaaag
     1921 agcaatttga acaatgggtt gctcgccttc aaatatccga ggcagggcaa aaaattattg
     1981 agaaagttcg ctcatctgag ccatcccgtc gcgttggtgg cggtagcaag aatgtttcgg
     2041 gacgctaccc tagtcgcaag atgggagtca cgattcaatt tgagtctcat cgggttgaac
     2101 tgccaattat ctatcagtta gaacatgata aagatgttct agaattctac gatcaaccac
     2161 cgcaaattaa acttgactat caagcaagta acggacgtag attaggcgtt ttacacaccc
     2221 cagatttatt tgtcatccga accaattcgg ctggttggga agagtgtaaa actgagcagg
     2281 atttgaaaaa actagcagag aaaaacccca atcgttactt ttactctgaa gaaaataatc
     2341 aatggcattc cttgccagga gaagtttatg ctaaccagtt tgggctttat caccgcattc
     2401 attcagatcg cgaaatcaat tgggttctgc aacgtaattt gcaattttta gaagattatt
     2461 atcgctcaga atctctagta gtggaagaag cgatcgctca atcattactt gcaattgtgt
     2521 catctcaacc aggaatcact ttagcagaat tactcaatcg gtcaacagga gctaaatctg
     2581 acgatatcta caccttaatt attcaagagc aaatctatat tgatttaaat gctacaccac
     2641 tagctgaacc agaaagatgt ctaatcttcc gtgacgagca aactgctagt gcttatagat
     2701 taatggttga gcaaccaagt gtaagtcttc cggctatctc tcctgtagtc aatattgtta
     2761 ctggagcaac tgttagttgg gacggcaaag gtttaaatat catccatgtt ggagaaacag
     2821 aggttatact tggcgcagaa aataatcagt taattgaact caaaaaagct atatttgaga
     2881 atctggttcg ccaaggaaaa atcacaagtc tccaaacacc agaaaaaaca gctattagca
     2941 cagaatcttg gcagcgattt catcaagcta gtcctgaaga ccaagcagaa gcccttgaac
     3001 gctacaaaac tatcgaacct tacttaaacg gacatcctcc agaaaacgaa acaataccag
     3061 cccgtaccat tcgtcactgg aaagctaaat atttaattgc aattcaaaaa tacggttgcg
     3121 gctacattgg gcttttgtcc catcgcagtg tcaaaggtaa tcgacagcgc aagctgcccg
     3181 aagacacctt agcaattatg gagaagttta ttttagaaga ctatgaaact cttaaacaaa
     3241 aaagaatgtg ggaggttcac gcggctttag ttcgagcttg cgaagaagct ggtgtaattg
     3301 ctccgagcta taaagcattt accaaagaag tacaacgccg gactggttac gaacagacta
     3361 aaaaacgcca aggtcgtcgt gccgcttatc agcatgaatc cttctattgg gagcttgcta
     3421 taactacccc aagacacggc gatcgccctt ttgaaattgg gcatatagat catactgagt
     3481 tagatgtaga gctagtttgc tctgatacag gtcgcaatct tggcagacct tgggcaacat
     3541 ttctcgtaga tgcttattca cggcgattgc tggcagttta cctaacgttt gactcaccct
     3601 catatcgttc ttgcttaatg gtgctaagaa tttgtgtcaa gcgtcatggc aggcttcccc
     3661 aaattgtagt ggtagataac ggtgcagagt ttcacagcgt ttactttgaa acactactag
     3721 ctacttttga atgtacaaaa aagcaaagac ccccagccaa agctcgtttt ggttcggttt
     3781 gcgaacgcct atttggaact tccaatactc aatttgtcca taacttactt ggtaatacac
     3841 aaatcactcg taatgtccgc caagtcacta agtcagttaa ccctaaaaat ctagctgtat
     3901 ggacacttgg tttactttat gaatatcttt gtgcttgggc ttacgaagtt tatgatacag
     3961 acgaacatcc ggcacttttc cagagtccga gggatgcgtt tgctgcgggt atggcgattg
     4021 gaggcagtcg ggttcaccgc atgattcctt acgatgaaaa ctttcagatt cttaccctac
     4081 ctacaacctc agagggaaaa gcaaaagttc aagttggacg cggcataaaa attaactaca
     4141 tatactactg gtctaatagt ttccgcgatc ctcaaattga aaatacctct gtgcagataa
     4201 gatacgaccc gtttaatatt ggcatcgctt acgcctttgt tcgaggacag tgggtacagt
     4261 gtatctctca atattatgca gaactccaag gtcgttcaga aaaagaactg aagttagcta
     4321 gtatcgagtt acgcaaacac tattcaaatc atacacaaca atctaaagtt tcggctaaga
     4381 atttagcaga atttttagca tcggtagaag ctcaagaagc tctattagaa cagcgttcct
     4441 atgatgcaga agttaaggaa gtctttaggg taatcgaggg tggaaaagcg acaacaagcc
     4501 gtaacgaaga accaaaactt attcaagtta cttttgccaa taccgatttt caagcggatg
     4561 aagatgaagc gatcgtccca gaaacacttg tagtgtatga ggagttttaa ctatgacaaa
     4621 atctactgga tttccgcttg aactcctcac acggcctgct acagaaaggc ttgcttactt
     4681 tgaaaattac accgtagcgc atcctcgact taaagaagtc tatgaaattc tcatgcgaac
     4741 gattgccgaa ccagcaggag cttcatttat ctttgtttac ggtgctagtg gtgttggtaa
     4801 aacaactttg cgactccgcg ttgagcagaa attaacagaa ctagcattac ctaaattgga
     4861 aagcgatcgc gctcgtgttc ctgttgttgg tattgaagct atagcgccag aatcgagata
     4921 cttcaactgg aaagaatact acacacgcgc cttaattact ttggaagaac cattaatcga
     4981 tcacaagttt gattacggtg tgcgtggtat ctctcgcgat aattttggca aaattaatgt
     5041 tgagtcgaaa gtagttgcac ctgctctacg cagagcttta gaaaatgctc tcatccatcg
     5101 tcatccagac gtattttttg tagatgaagc acaacatttt ggtaaggtgg caagcggata
     5161 caaactacaa gatcaattag actgtctcaa atctttagca aatatgacag gcatcttgca
     5221 ctgtctatta ggaacatacg aactacttac attccgtaat ttgagcggtc aactctcacg
     5281 tcgtagtgtt gatattcatt tccgccgtta ttgtgctgat agcccagaag atgtacaggc
     5341 ttttaagagt gtgttgttaa cttttcaaca gcatctgcca ctagcagaaa ctccaaattt
     5401 agtcgatcat tgggaatatt tttacgaacg tactttagga tgtattggca cactgaaaga
     5461 ttggctgaag cgtgttctta gtgatgcttt ggatcgagaa gcaacaacaa tcactctcaa
     5521 agatttacaa aaacgtgctt tgtcggttgc tcagtgtcag aaaatgttca aggaaattca
     5581 agagggtgaa cgacaattat ctgaaacaga agcagatgtg caaaacttac gctctgcatt
     5641 aggacttggc gcaaaaccaa ttgtgctacc cgaagaaacc ccaaaaacta ctcgtccgcc
     5701 aggaaaggtt ggcaaacgca aacctaaaag agatcccatc ggagtgcagc aagatgttag
     5761 ctagcttaag tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc gaaattaata
     5821 cgactcacta taggggaatt gtgagcggat aacaattccc catcttagta tattagttaa
     5881 gtataagaag gagatataca tatggcagat ctcaattgga tatcggccgg ccacgcgatc
     5941 gctgacgtcg gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag
     6001 cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa
     6061 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaacct
     6121 caggcatttg agaagcacac ggtcacactg cttccggtag tgccctgaac cgacgaccgg
     6181 gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc
     6241 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     6301 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     6361 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     6421 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     6481 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     6541 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     6601 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     6661 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     6721 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     6781 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     6841 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     6901 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     6961 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     7021 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7081 atacatattt gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc
     7141 tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt
     7201 ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc
     7261 cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg
     7321 ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag
     7381 atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt
     7441 taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca
     7501 cacactggta tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac
     7561 tccccgttca gcccga
//
