LOCUS       sequence-418053-        5003 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GstTnpB2 and native omega-RNA from constitutive promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5003)
  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 5003)
  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: pSL4369 (pEffector_TnpB2,omega-RNA)
FEATURES             Location/Qualifiers
     source          1..5003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      134..872
                     /direction=RIGHT
                     /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."
     promoter        920..1011
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1012..1803
                     /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"
     terminator      complement(1974..2021)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2017..2035
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(3336..3370)
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     terminator      complement(3382..3429)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(3727..3761)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        3767..3844
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3845..4927
                     /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(3864..3883)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ccagtcagct ccttccggtg ggcgcggggc atgactaaca tgagaattac aacttatatc
       61 gtatggggct gacttcaggt gctacatttg aagagataaa ttgcactgaa atctagagcg
      121 gttcagtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctttt
      181 gccctgtaaa cgaaaaaacc acctggggag gtggtttgat cgaaggttaa gtcagttggg
      241 gaactgctta accgtggtaa ctggctttcg cagagcacag caaccaaatc tgtccttcca
      301 gtgtagccgg actttggcgc acacttcaag agcaaccgcg tgtttagcta aacaaatcct
      361 ctgcgaactc ccagttacca atggctgctg ccagtggcgt tttaccgtgc ttttccgggt
      421 tggactcaag tgaacagtta ccggataagg cgcagcagtc gggctgaacg gggagttctt
      481 gcttacagcc cagcttggag cgaacgacct acaccgagcc gagataccag tgtgtgagct
      541 atgagaaagc gccacacttc ccgtaaggga gaaaggcgga acaggtatcc ggtaaacggc
      601 agggtcggaa caggagagcg caagagggag cgacccgccg gaaacggtgg ggatctttaa
      661 gtcctgtcgg gtttcgcccg tactgtcaga ttcatggttg agcctcacgg ctcccacaga
      721 tgcaccggaa aagcgtctgt ttatgtgaac tctggcagga gggcggagcc tatggaaaaa
      781 cgccaccggc gcggccctgc tgttttgcct cacatgttag tcccctgctt atccacggaa
      841 tctgtgggta actttgtatg tgtccgcagc gcccgccgca gtctcacgcc cggagcgtag
      901 cgaccgagtg agctagctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      961 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagggaa
     1021 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     1081 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     1141 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     1201 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     1261 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     1321 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     1381 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     1441 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     1501 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     1561 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     1621 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     1681 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     1741 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     1801 taatgtctaa caattcgttc aagccgaggg gccgcaagat ccggccacga tgacccggtc
     1861 gtcggttcag ggcagggtcg ttaaatagcc gcttatgtct attgctggtt taccggttta
     1921 ttgactaccg gaagcagtgt gaccgtgtgc ttctcaaatg cctgaggttt cagcaaaaaa
     1981 cccctcaaga cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca
     2041 gcagcctagg ttatcacgcg atgcgaatac gttgcggggt gtctacctca ccagaactgc
     2101 acaagacaga caaatcaaca gccttgcttt tttttaaaat attcaatgcc ccattcacgt
     2161 ctgcgttaat aagttttcct gatttagtgc gatacaggcc gcgcttaatg cgcttaccag
     2221 aaaatttgta ctctttggga ttatccgcgt tgtattcggg gatttcatct ccatcaaaaa
     2281 acgacgcttg acttgtatac gactcttctt gcttcaaaaa ctcgatccca tagaactcac
     2341 aaaggtattc taacttctcc ttaatgtttc caagagggat attgacgaag ttttggttgg
     2401 ttttcttgcc caaattcatg ttgcgctgcc atgtttcggc gtacccgatt accagcttgc
     2461 caatttgatt ttcgatacag tagttaatga tatagcggca ggtcttattg atatagtcgt
     2521 tcaccttgtt attacgattc atcgcaagca gtgcttgttt acgcgtggtt cccttaatct
     2581 tctgtttatc tttaatagac tgcaagcgag cgttttcctt attgaaccat tggttgatag
     2641 atttaagacg acggccgtca ataatgaaag aacgtccgtc tgaggttaca catgtagcca
     2701 agttattaag ccccaagtca atcgcaagcg ccttctgatc attaagttcg cgctgatctt
     2761 caggcatttc gtacttgtac tggatctcga agaaacgtgc atggtgtttc ggaatgattt
     2821 cgatctgctt aatcttttta tcaagtaaca ctggtggaat agtgattgta atgggcttat
     2881 gtgtcttttt gaataagcga gagtatggaa tcgtgaactt atttccgtca atgcggattt
     2941 ggccgatgat taacgaatgg aaaccgtctt tctttaagta tttgggaatc gagattgctt
     3001 tatggtcgta tttaccttgt ttggcaagcg aaataagacc gaagaaactt ttgaaggctt
     3061 catttacttt tttcaagatt tgttgagcca tgttcgagtt cagtaacttg tagttttcat
     3121 tagtcttcgc caagtgatag ttcttttcgt aatttaagaa ctccttatgc tcaaaatagt
     3181 actgacgtac gttgtaaagg ccaacgttgt acatgttttt ggcaatatgg cacaactcac
     3241 gcagtgtcag gtactcttcc ttcgtcaacc cattaagctg ctgcttgatg caaaaatagc
     3301 ccatggtata tctccttggt accgtatatc tccacgctag catagtacct aggactgagc
     3361 tagccgtaaa atcgacttca gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     3421 ttatgctagt tattgcggcc gcgagctatc ataccataaa gcaacgcatg gtgaaccact
     3481 tgtccgtgaa ctacccccac ttaattttct agcgaaaatt tgaagtgggg gcttccaaag
     3541 aagtttgact gcttcaagca atccttattc tttgaggcgt gtccacttcg ccgctagagc
     3601 ataagacact caggtctaca gctttacttt tctttaagat gtttaatgcg ccattgacat
     3661 cagcattaat tagtttgcca gactttgttc gatacaagcc gcgcttaact cgaggtatat
     3721 ctccacgcta gcattatacc taggactgag ctagctgtca atcccggaca ccatcgaatg
     3781 gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt
     3841 gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac
     3901 cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga
     3961 agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa
     4021 acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat
     4081 tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt
     4141 agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt
     4201 cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc
     4261 ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat
     4321 tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca
     4381 ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc
     4441 tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga
     4501 ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc
     4561 cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga
     4621 gtccgggctg cgcgttggtg cggacatctc ggtagtggga tacgacgata ccgaagacag
     4681 ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac
     4741 cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt
     4801 gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc
     4861 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4921 gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac
     4981 cgatgccctt gagagccttc aac
//
