LOCUS       sequence-418055-        4869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GstTnpB4 and native omega-RNA from constitutive promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4869)
  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 4869)
  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: pSL4673 (pEffector_TnpB4,omega-RNA)
FEATURES             Location/Qualifiers
     source          1..4869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(143..177)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        183..260
                     /gene="lacI"
                     /label=lacI promoter
     CDS             261..1343
                     /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(280..299)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      1553..2291
                     /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        2339..2430
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2431..3222
                     /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(3393..3440)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3436..3454
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(4623..4657)
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     terminator      complement(4670..4717)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 atcagggggc aggttgacaa ctacccaacg acgcaggtca tgcctgtgtc gttccttcaa
       61 cgatgtactc tattcctttt cgttgaaggt tcatagcacc aatgcggtca tcgtttgagg
      121 tatactcgag gtatatctcc acgctagcat tatacctagg actgagctag ctgtcaatcc
      181 cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
      241 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      301 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
      361 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
      421 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
      481 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
      541 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
      601 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
      661 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
      721 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
      781 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
      841 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
      901 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
      961 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     1021 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg
     1081 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     1141 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     1201 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     1261 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1321 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     1381 gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     1441 gcgcggggca tgactaacat gagaattaca acttatatcg tatggggctg acttcaggtg
     1501 ctacatttga agagataaat tgcactgaaa tctagagcgg ttcagtagaa aagatcaaag
     1561 gatcttcttg agatcctttt tttctgcgcg taatcttttg ccctgtaaac gaaaaaacca
     1621 cctggggagg tggtttgatc gaaggttaag tcagttgggg aactgcttaa ccgtggtaac
     1681 tggctttcgc agagcacagc aaccaaatct gtccttccag tgtagccgga ctttggcgca
     1741 cacttcaaga gcaaccgcgt gtttagctaa acaaatcctc tgcgaactcc cagttaccaa
     1801 tggctgctgc cagtggcgtt ttaccgtgct tttccgggtt ggactcaagt gaacagttac
     1861 cggataaggc gcagcagtcg ggctgaacgg ggagttcttg cttacagccc agcttggagc
     1921 gaacgaccta caccgagccg agataccagt gtgtgagcta tgagaaagcg ccacacttcc
     1981 cgtaagggag aaaggcggaa caggtatccg gtaaacggca gggtcggaac aggagagcgc
     2041 aagagggagc gacccgccgg aaacggtggg gatctttaag tcctgtcggg tttcgcccgt
     2101 actgtcagat tcatggttga gcctcacggc tcccacagat gcaccggaaa agcgtctgtt
     2161 tatgtgaact ctggcaggag ggcggagcct atggaaaaac gccaccggcg cggccctgct
     2221 gttttgcctc acatgttagt cccctgctta tccacggaat ctgtgggtaa ctttgtatgt
     2281 gtccgcagcg cccgccgcag tctcacgccc ggagcgtagc gaccgagtga gctagctatt
     2341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2401 atgcttcaat aatattgaaa aaggaagagt atgagggaag cggtgatcgc cgaagtatcg
     2461 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     2521 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     2581 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     2641 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     2701 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     2761 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     2821 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     2881 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     2941 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     3001 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
     3061 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
     3121 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
     3181 cactacgtga aaggcgagat caccaaggta gtcggcaaat aatgtctaac aattcgttca
     3241 agccgagggg ccgcaagatc cggccacgat gacccggtcg tcggttcagg gcagggtcgt
     3301 taaatagccg cttatgtcta ttgctggttt accggtttat tgactaccgg aagcagtgtg
     3361 accgtgtgct tctcaaatgc ctgaggtttc agcaaaaaac ccctcaagac ccgtttagag
     3421 gccccaaggg gttatgctag ttattgctca gcggtggcag caggagctct tatcacgctt
     3481 gggtggtgcc ttcaacaata tattcaatcc ccttacgttg caagttcatg gccccgatac
     3541 ggtcatcgtt gctcgtgtag ccgcaagtgc gacaacagaa ggtgtgggtg cgctttttac
     3601 ggttggcttt ttctgtatgt ccacatttgg ggcaggtcaa ggatgtgtaa tgaggagcga
     3661 cgacgataac cttggaccca tgcaagcgtg ccttgtactc taacatttga cggaattgat
     3721 aaaatgccca agacaccgtc tcgtaacggt cgtggatgcg cacacgttct gttttctcgc
     3781 ggatccccgt aaggtcttcc aaaacgaaca acgtgcgttc gccgtattga cgcactaatg
     3841 ctttagtaac tgcatggttc acgtcagtca tccagcgatt ctcacgttgg ccgatggttt
     3901 tcagcttgcg acgtgccgag gccgttccct tttgctgaag cgttttgcgc atacgtttgt
     3961 acttagcgcg catgtgttta atcttacgac cgttaaaaaa aatggttttt ccttgcgagt
     4021 cataagcaac agccaagaag ttcacaccca catccacccc aacgacctgg cgaatattgt
     4081 gttcatccac ggtaggaatt gttttagtca tcgggatgtg cagaaaaaat ttgttatgtt
     4141 tgtaaaccag cttggccgtt ccaaaagtcc aggacccgtc aaaataaggt tccatacctt
     4201 tcggttcaaa tgagaccttg atgcgtcctt ccagggtgtt gacggaaaaa aggccttgga
     4261 caatggagta gtcacgattc caaaccaagt cgtattctgg tttcttgaag cgtaccaagg
     4321 tccactcatg gccattagac tttaacgaac gataacgagc gataactgtt ttgatcaccg
     4381 attgggccat ttgtgaacgc agtgcgtagt tagaacgcag taagcggtat gtcatgtcat
     4441 gcagttttgc ttgggccagc acctttgtgt caaaaattac tactgaaacg gcattacagg
     4501 cttgacggta agctgataat gtagccttta atacttcagc ctgttccgca gtagggtaaa
     4561 ttttgatttt tgcggtcagc gtcatatcca tggtatatct ccttggtacc gtatatctcc
     4621 acgctagcat agtacctagg actgagctag ccgtaaaagt cgacttcagc aaaaaacccc
     4681 tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgcggccgc gagtaaggaa
     4741 ccggaccaat ttgcacaaca aacaagaggg gcggtcaact acccccactt agctaacgct
     4801 tgaagtgggg gcttgcaact ccccagaagt gcgagcggac ttcgtcctcc ttccttgact
     4861 tggggttgc
//
