LOCUS       sequence-418054-        4932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GstTnpB3 and native omega-RNA from constitutive promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4932)
  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 4932)
  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: pSL4672 (pEffector_TnpB3,omega-RNA)
FEATURES             Location/Qualifiers
     source          1..4932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1212..1230)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1226..1273
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1444..2235)
                     /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(2236..2327)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2375..3113)
                     /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(3323..4405)
                     /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     4367..4386
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4406..4483)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4489..4523
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     terminator      4818..4865
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        4878..4912
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
ORIGIN
        1 ggagatatac catgggctac cgcacactga aaactcgttt ccgcgccaag aaagaagtta
       61 tccagaagtt gtttgaatgc aaccgtatct ccgcggaggt ttggaacgag tgcctgcgct
      121 tagctaaaga gcatcacctt gagaccggca agtggattac taaaacggaa cttcaaaaag
      181 ccacgaaggg ccgcttttct attcatagcc agagtattca agcggtagtt cataagtaca
      241 ttttcgcgcg cgatggggct aaagaggccc gtaagaaagg agaaaaaatc aagtatccat
      301 acaaaaaaaa gaagcacttc aatactaagt gggctaaaga tggcttcgtg ttgcatgaag
      361 acggaacgct tgaattgtcg ttaggaatct ggaatcgtaa gcgtcaatcg cctttggtgg
      421 tgaaaatcga taaagagaag ttaccaaacg gcaaagtaaa agaaattgag ttggtctatg
      481 accgcgggtt gtggttatgc ttgagctatg aagacggcaa aaaacctaaa gaaaaccctt
      541 gtaagaatcg cgtagcaatc gacccaggag agatccacac gattgcagcg atctgtgaga
      601 atggcgaatc gctgattatt actggtcgta aaatccgctc gattcaccgc cttcgtaaca
      661 agaaacttaa agaattgcag aagttgatgt cgcgctgtaa gaaggggagt aagcagtgga
      721 aaaaatataa ccgtgccaag cagtatgttt tatcaaaatc ggaagcccag cttaaagatg
      781 ctttgcacaa aacgacccgc caattcgttc gctggtgtct ggagaatcaa gtcaaggagg
      841 tggtaattgg tgacatcgag ggtattcagc gtaatactaa gaagaagcgc aacaagaaaa
      901 cgaatcaaaa gctgagtaat tggtcgttcg gtaagctgtt tgattatctg aagtataagt
      961 taaatgccga ggggatccag attgaaaaaa aggacgaatc gtacacctca caaacctgcc
     1021 cggtctgcgg aaagaagaac aagtcgagtt cacgcaacta cacctgtcag tgtgggtata
     1081 aacgtcaccg tgatatccac ggggccatga atttattcgc aaaggtctat tacggggaga
     1141 ttcgccccct tgagtttacc gttaaaccgt tcacttaccg tcgtatcgcg taataagagc
     1201 tcctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1261 gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     1321 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     1381 accgacgacc gggtcatcgt ggccggatct tgcggcccct cggcttgaac gaattgttag
     1441 acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa attcttccaa
     1501 ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct gtctagcttc
     1561 aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg cagcgacatc
     1621 cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg taagcactac
     1681 atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg tttcatttag
     1741 cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg ctggacctac
     1801 caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa tgtcgatcgt
     1861 ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa attgcagttc
     1921 gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg tgacttctac
     1981 agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt cgttgatcaa
     2041 agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat caatatcact
     2101 gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca gcaacgtcgg
     2161 ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata cttcggcgat
     2221 caccgcttcc ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2281 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag ctagctcact
     2341 cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata caaagttacc
     2401 cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag ggccgcgccg
     2461 gtggcgtttt tccataggct ccgccctcct gccagagttc acataaacag acgcttttcc
     2521 ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg cgaaacccga
     2581 caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct ctcctgttcc
     2641 gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag tgtggcgctt
     2701 tctcatagct cacacactgg tatctcggct cggtgtaggt cgttcgctcc aagctgggct
     2761 gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac tgttcacttg
     2821 agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt aactgggagt
     2881 tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc caaagtccgg
     2941 ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac cacggttaag
     3001 cagttcccca actgacttaa ccttcgatca aaccacctcc ccaggtggtt ttttcgttta
     3061 cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3121 ctgaaccgct ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc
     3181 ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct
     3241 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     3301 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3361 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3421 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     3481 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     3541 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     3601 agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     3661 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     3721 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     3781 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     3841 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     3901 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     3961 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     4021 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     4081 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     4141 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     4201 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     4261 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     4321 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4381 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     4441 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatt gacagctagc
     4501 tcagtcctag gtataatgct agcgtggaga tatacctcga gtttacggtc aaaccgttta
     4561 cgtatcgacg gattgcttag gtatgaagtc gtagatggcg agttcccgcc cttgagtatc
     4621 atggatactc atgttgcctg tggctcccaa cgggacgggt gccaggtgtt gcctgcagat
     4681 aggctgccaa ccagcctatc atgcatccac ctccgaaggg gaaacaggaa acccccactt
     4741 cgataagtgg gggaggttca tcggtcattt tcccgtgttt tcgttctctg tctccagcgc
     4801 actcgcggcc gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     4861 ttttgctgaa gtcgactttt acggctagct cagtcctagg tactatgcta gcgtggagat
     4921 atacggtacc aa
//
