LOCUS       sequence-418052-        4862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Harbors a native ISGst3 transposon derived form Geobacillus 
            stearothermophilus DSM 458.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4862)
  AUTHORS   Meers C, Le HC, Pesari SR, Hoffmann FT, Walker MWG, Gezelle J, Tang 
            S, Sternberg SH
  TITLE     Transposon-encoded nucleases use guide RNAs to promote their selfish
            spread.
  JOURNAL   Nature. 2023 Sep 27. doi: 10.1038/s41586-023-06597-1.
  PUBMED    37758954
REFERENCE   2  (bases 1 to 4862)
  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"; journalName: "Nature. 
            2023 Sep 27. doi: 10.1038/s41586-023-06597-1."
COMMENT     Sequence Label: pSL5200 (pDonor_ISGst3)
FEATURES             Location/Qualifiers
     source          1..4862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(228..246)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      242..289
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(460..1251)
                     /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(1252..1343)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1391..2129)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(2339..3421)
                     /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     3383..3402
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3422..3499)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        3510..3528
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tggcgcatta aacatcttaa agaaaagtaa agctgtagac ctgagtgtct tatgctctag
       61 cggcgaagtg gacacgcctc aaagaataag gattgcttga agcagtcaaa cttctttgga
      121 agcccccact tcaaattttc gctagaaaat taagtggggg tagttcacgg acaagtggtt
      181 caccatgcgt tgctttatgg tatgatagag cgcctaggct gctgccaccg ctgagcaata
      241 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaacctca
      301 ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca
      361 atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt catcgtggcc
      421 ggatcttgcg gcccctcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt
      481 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
      541 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
      601 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
      661 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
      721 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
      781 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
      841 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
      901 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
      961 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     1021 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     1081 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatccac
     1141 tgcggagccg tacaaatgta cggccagcaa cgtcggttcg agatggcgct cgatgacgcc
     1201 aactacctct gatagttgag tcgatacttc ggcgatcacc gcttccctca tactcttcct
     1261 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1321 atgtatttag aaaaataaac aaatagctag ctcactcggt cgctacgctc cgggcgtgag
     1381 actgcggcgg gcgctgcgga cacatacaaa gttacccaca gattccgtgg ataagcaggg
     1441 gactaacatg tgaggcaaaa cagcagggcc gcgccggtgg cgtttttcca taggctccgc
     1501 cctcctgcca gagttcacat aaacagacgc ttttccggtg catctgtggg agccgtgagg
     1561 ctcaaccatg aatctgacag tacgggcgaa acccgacagg acttaaagat ccccaccgtt
     1621 tccggcgggt cgctccctct tgcgctctcc tgttccgacc ctgccgttta ccggatacct
     1681 gttccgcctt tctcccttac gggaagtgtg gcgctttctc atagctcaca cactggtatc
     1741 tcggctcggt gtaggtcgtt cgctccaagc tgggctgtaa gcaagaactc cccgttcagc
     1801 ccgactgctg cgccttatcc ggtaactgtt cacttgagtc caacccggaa aagcacggta
     1861 aaacgccact ggcagcagcc attggtaact gggagttcgc agaggatttg tttagctaaa
     1921 cacgcggttg ctcttgaagt gtgcgccaaa gtccggctac actggaagga cagatttggt
     1981 tgctgtgctc tgcgaaagcc agttaccacg gttaagcagt tccccaactg acttaacctt
     2041 cgatcaaacc acctccccag gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa
     2101 aaaaaggatc tcaagaagat cctttgatct tttctactga accgctctag atttcagtgc
     2161 aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct
     2221 catgttagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     2281 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     2341 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2401 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     2461 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     2521 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     2581 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     2641 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     2701 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     2761 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     2821 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     2881 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     2941 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     3001 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     3061 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     3121 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     3181 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     3241 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     3301 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     3361 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     3421 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     3481 tgcgccattc gatggtgtcc gggagaaatt aatacgactc actataggcc tcgagattgg
     3541 cggagcgggc ctcgtaaaag gctcgttttt tttatcttaa ttaggcgaga agactcccac
     3601 ttcaacgaag cgaagcggag taagtgggag atgaatcgcc ttaactatat tttgctgaaa
     3661 ataggataga ttcagtaaaa tatagtatca tatagttaga tgggaggtga aaacatgtat
     3721 ttttgtatca aacaacagct aaatggtttg accaaagaag aatacttgac tcttcgagaa
     3781 ctgtgccata ttgccaagaa catgtacaac gtcggattgt acaatgtcag acaatactat
     3841 tttgaacaca aggaatttct taattatgag aaaaactatc atcttgcaaa aactaacgaa
     3901 aactataagc tgttaaacag caacatggca cagcaaattt taaaaaaggt taatgaagcc
     3961 tttaaatctt tctttggttt gatcagtctt gccaaacaag gaaaatatga ccacaaggct
     4021 atcagtattc caaaatatct taaaaaagat ggctttcatt cactgatcat tggccagatt
     4081 cgtatagacg gcaacaaatt cacgataccg tattctcgcc tatttaaaaa gactcacaag
     4141 cctatcacga taacgattcc gcctgtgtta ctggacaaaa agattaagca gattgaaatc
     4201 attcctaagc atcatgccag gttctttgag attcagtaca aatatgaaat gcctgaagat
     4261 caaagagaat taaacgacca aaaagcactg gcaattgatt taggattaaa caatcttgcc
     4321 acttgtgtca catcagacgg cagatcattc atcattgatg ggcggagatt aaaaagtata
     4381 aatcaatggt ttaacaaaga aaatgccaga cttcaaagca taaaagataa gcaaaaaatc
     4441 aaaggcacca ctcgtaaaca ggctttgctt gctatgaatc gcaataataa agtgaatgat
     4501 tatatcaaca agacttgccg ttacatcatt aactactgta ttgaaaatca aattggcaaa
     4561 cttgtcattg gctatgcgga aacatggcaa cgcaatatga atctaggaaa aaagacaaat
     4621 caaaactttg tcaatattcc tctcggtaac ataaaagaaa aactagaata tctttgtgaa
     4681 ttttacggca ttgaattctt gaaacaggaa gaatcctata cgtctcaagc cagctttttt
     4741 gacggcgatg agattcctga atataatgcc gacaatccaa aagaatataa gttcagcggc
     4801 aaacgtatta agcgcggctt gtatcgaaca aagtctggca aactaattaa tgctgatgtc
     4861 aa
//
