LOCUS       sequence-418056-        5023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GstTnpB1 and native omega-RNA from constitutive promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5023)
  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 5023)
  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: pSL4624 (pEffector_TnpB1,omega-RNA)
FEATURES             Location/Qualifiers
     source          1..5023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(463..481)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        492..569
                     /gene="lacI"
                     /label=lacI promoter
     CDS             570..1652
                     /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(589..608)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      1862..2600
                     /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        2648..2739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2740..3531
                     /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(3702..3749)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3745..3763
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     regulatory      4887..4896
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4916..4940
                     /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(4941..4959)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(4972..5019)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tgcggccgcg agttcccagt acacagaagg cagttcatgc ttttcgatca gcgtgacata
       61 ctcccaccac ctacgcttcg cttagaggtg ggggcttctc gggtaatccc atctcatgat
      121 gggaagttga ccgagcgatc cccgtgtgcc ccacggttcg aggacaaatt agacgatcgc
      181 caatcgtttc atgccctcat gtttgatatt gatggccgca ttgacgtccc tgtcgctctc
      241 gaatccacat tcacagcgga atgtccgttc agaaagcgat agagactcct tgactcgacc
      301 gcagcacgaa cacgttttgg acgaggggaa ccatttgtcg attttgatca gcttcttccc
      361 ctgttcggcc agcttgtact gaaggaaagt ggtgaacatg ccccagccgt tgtcatgaac
      421 gccttgaccg aaatggaggg cttgtgacat ccttactcga ggcctatagt gagtcgtatt
      481 aatttctccc ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
      541 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
      601 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
      661 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
      721 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
      781 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
      841 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
      901 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
      961 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     1021 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
     1081 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     1141 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     1201 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     1261 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     1321 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggac atctcggtag
     1381 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac
     1441 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
     1501 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg
     1561 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     1621 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc
     1681 actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     1741 ttccggtggg cgcggggcat gactaacatg agaattacaa cttatatcgt atggggctga
     1801 cttcaggtgc tacatttgaa gagataaatt gcactgaaat ctagagcggt tcagtagaaa
     1861 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatcttttgc cctgtaaacg
     1921 aaaaaaccac ctggggaggt ggtttgatcg aaggttaagt cagttgggga actgcttaac
     1981 cgtggtaact ggctttcgca gagcacagca accaaatctg tccttccagt gtagccggac
     2041 tttggcgcac acttcaagag caaccgcgtg tttagctaaa caaatcctct gcgaactccc
     2101 agttaccaat ggctgctgcc agtggcgttt taccgtgctt ttccgggttg gactcaagtg
     2161 aacagttacc ggataaggcg cagcagtcgg gctgaacggg gagttcttgc ttacagccca
     2221 gcttggagcg aacgacctac accgagccga gataccagtg tgtgagctat gagaaagcgc
     2281 cacacttccc gtaagggaga aaggcggaac aggtatccgg taaacggcag ggtcggaaca
     2341 ggagagcgca agagggagcg acccgccgga aacggtgggg atctttaagt cctgtcgggt
     2401 ttcgcccgta ctgtcagatt catggttgag cctcacggct cccacagatg caccggaaaa
     2461 gcgtctgttt atgtgaactc tggcaggagg gcggagccta tggaaaaacg ccaccggcgc
     2521 ggccctgctg ttttgcctca catgttagtc ccctgcttat ccacggaatc tgtgggtaac
     2581 tttgtatgtg tccgcagcgc ccgccgcagt ctcacgcccg gagcgtagcg accgagtgag
     2641 ctagctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2701 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagggaagc ggtgatcgcc
     2761 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     2821 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     2881 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     2941 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     3001 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     3061 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     3121 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     3181 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     3241 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     3301 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     3361 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     3421 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     3481 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctaaca
     3541 attcgttcaa gccgaggggc cgcaagatcc ggccacgatg acccggtcgt cggttcaggg
     3601 cagggtcgtt aaatagccgc ttatgtctat tgctggttta ccggtttatt gactaccgga
     3661 agcagtgtga ccgtgtgctt ctcaaatgcc tgaggtttca gcaaaaaacc cctcaagacc
     3721 cgtttagagg ccccaagggg ttatgctagt tattgctcag cggtggcagc agcctaggtt
     3781 atcatacaat ggccaaacgc ttcatcccct cgtgcttaat gttgatagcc gcgtttacat
     3841 cgcggtcact ttcgaacccg cactcacaac ggaaggtgcg ttcgctcagt gacaagctct
     3901 ctttcacacg cccgcaacag ctgcaggttt ttgaggatgg gaaccactta tcaatcttaa
     3961 tcaacttttt cccttgctcc gccaatttat actgtaaaaa cgttgtaaac ataccccatc
     4021 cattatcatg taccccctgt ccaaagtgca gcgcttggct catgcctttc atattaagat
     4081 cctcgattac aacgcaatca tagcgcttgg caagtttgtg agattccttg tgcagaaagt
     4141 ccttgcgttg gttcgccatc ttttcgtgca atttggcaac ctttaaacgc tgcttatgcc
     4201 aacgatttga tccttttttt ttacgagaaa gcttacgttg ttctttggcc aacgtttcca
     4261 atgctttacg atagaagcgc ggatagttag cacgcttgcc ttcagaatca acatacaacg
     4321 tgctcatact gaaatccagt ccaaccacag tttgcacctc tttaagtgct ggttggtgct
     4381 cgtattctgt taagatgcta atgtagtatt tcccggtctt tgtcttagtg atagtgcaac
     4441 ttttgataat atgatgcgct ggaatttcac gatgctgttt aaatttgatc catttcagtt
     4501 ttggcaattt gatataccca tccgacaact taatgttgcc attcaccatg tttgtagtgt
     4561 atgattgttt cgccttacgg ttcttaaatt tagggaatcc ggcacggccg gagaaaaagt
     4621 gttgaaatgc tttttgcagg tttaattggg cgttggccag tgccaagctg tccacttctt
     4681 tcaaccaagg aaattccttt ttatatttag caggtgtcgg acacgtttgt tgttttaacg
     4741 attcttgatc atccttgaac ttctcgaaaa gctgaatacg ttcttccagc attttgttgt
     4801 aaacaaagcg cacacagccg aaagttttcg ccagaagttg ctcttgttct ttggttggat
     4861 aaagacggaa ttggtaggct ttgttggcca ccatggtata tctccttggt acctgggaat
     4921 tgttatccgc tcacaattcc cctatagtga gtcgtattaa gtcgacttca gcaaaaaacc
     4981 cctcaagacc cgtttagagg ccccaagggg ttatgctagt tat
//
