LOCUS       sequence-418057-        5048 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses GstIscB and native omega-RNA from constitutive promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5048)
  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 5048)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pSL4164 (pEffector_IscB,omega-RNA)
FEATURES             Location/Qualifiers
     source          1..5048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      87..824
                     /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        872..963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             964..1755
                     /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(1926..1973)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1969..1987
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    3290..3314
                     /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(3315..3333)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(3346..3393)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(3736..3754)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3765..3842
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3843..4925
                     /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(3862..3881)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tacaacttat atcgtatggg gctgacttca ggtgctacat ttgaagagat aaattgcact
       61 gaaatctaga gcggttcagt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      121 cgcgtaatct tttgccctgt aaacgaaaaa accacctggg gaggtggttt gatcgaaggt
      181 taagtcagtt ggggaactgc ttaaccgtgg taactggctt tcgcagagca cagcaaccaa
      241 atctgtcctt ccagtgtagc cggactttgg cgcacacttc aagagcaacc gcgtgtttag
      301 ctaaacaaat cctctgcgaa ctcccagtta ccaatggctg ctgccagtgg cgtttaccgt
      361 gcttttccgg gttggactca agtgaacagt taccggataa ggcgcagcag tcgggctgaa
      421 cggggagttc ttgcttacag cccagcttgg agcgaacgac ctacaccgag ccgagatacc
      481 agtgtgtgag ctatgagaaa gcgccacact tcccgtaagg gagaaaggcg gaacaggtat
      541 ccggtaaacg gcagggtcgg aacaggagag cgcaagaggg agcgacccgc cggaaacggt
      601 ggggatcttt aagtcctgtc gggtttcgcc cgtactgtca gattcatggt tgagcctcac
      661 ggctcccaca gatgcaccgg aaaagcgtct gtttatgtga actctggcag gagggcggag
      721 cctatggaaa aacgccaccg gcgcggccct gctgttttgc ctcacatgtt agtcccctgc
      781 ttatccacgg aatctgtggg taactttgta tgtgtccgca gcgcccgccg cagtctcacg
      841 cccggagcgt agcgaccgag tgagctagct atttgtttat ttttctaaat acattcaaat
      901 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      961 agtatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
     1021 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
     1081 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
     1141 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
     1201 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
     1261 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca
     1321 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
     1381 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
     1441 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
     1501 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
     1561 ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag
     1621 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg
     1681 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
     1741 gtagtcggca aataatgtct aacaattcgt tcaagccgag gggccgcaag atccggccac
     1801 gatgacccgg tcgtcggttc agggcagggt cgttaaatag ccgcttatgt ctattgctgg
     1861 tttaccggtt tattgactac cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggt
     1921 ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc
     1981 tcagcggtgg cagcagccta ggttatcacg ctaatgcgaa ggttggcgaa ctatccccct
     2041 tacggcgttc aatcagtaat gtacttgcgt gttcaacacg ttttaatttt ttccagctgg
     2101 ccgaagcgtg gactttagtc ccgtcaagca gacgtaaatc gaagtagccg gaagaacgac
     2161 gcccgaaaat aaaacactct ttacgctcat acacgacttt atcgaacagc tggaacccct
     2221 ttacgaagcg gggagccttg ttggatttac gcttaccgcc tttcaaaatt gtcgccttat
     2281 gtaattggcg attattcttg cgaacgaact tcagcaaata tacggtacca ctcggtttcg
     2341 ctaaaggatt gcctgaaatg caacgtgcgt ctaccatgtg cgacttgctt aacccgttcg
     2401 cgatgcgaat acatttggtg atataaccat atgtgatgtt cgcgtggctg tacttctcac
     2461 ggatgcggga aaacatcgcc caacgcatgg ccgtcatctg agacgcgtca cgcattggcg
     2521 ggcgtttgcg ttgaaaaata tgttccaggc ctttctcatg aatctcatga tggcacgtct
     2581 tgcacaaagt aatcaaatta tcgggcgagt cgccacccgt cttgcgagat tcaatatggt
     2641 gtacttgcaa tactggatct tttttcttac ccttacaata ctggcaggtg tggttgtcgc
     2701 gatataacac gtattcgcgt acgttccaaa aacccatttg ttccccctgt tggtactcga
     2761 cgccgcgaat gttcaagttt tttaatttct gggtgtcgaa cgcggctact tcaatagtaa
     2821 cagacgtcac gggcaaaatg ttatgaacta aatcaatcag cttaatatgc gaattaatct
     2881 tatgttggat tgaaggcggc aaccaccctt cgggtttacg acggttcaag aaacgagcct
     2941 cacggtaacg ggtcttgcgg ttgcggcgcg agcggcggaa ctggcggcgg atagctaaca
     3001 attccacaat atccgtacgc aactgcgcct ctgcctcgaa caatacatcc ttcttggtag
     3061 tagcagaaat gccgacatgg cgggtgccca tatccacgcc aagtgatacc ggctgcttgt
     3121 agccggagct gccatacagc aattgaatcg taaacggtgt acgacgaact acttttgctt
     3181 tcttttgttt cagaagcaga cgtgctttgc gggggctaca cggcattaac ggttttccat
     3241 gcttattgat aatgtataca aaccccatgg tatatctcct tggtacctgg gaattgttat
     3301 ccgctcacaa ttcccctata gtgagtcgta ttaagtcgac ttcagcaaaa aacccctcaa
     3361 gacccgttta gaggccccaa ggggttatgc tagttattgc ggccgcgagt ggttttccat
     3421 gcttattgat aatgtacacg aacaagacgt gtacctcctt tcaggagtga gttcctcctc
     3481 cccaatgttg gaagggcttg ttcggcagac agcaccgctc ctacccatca gagcttttaa
     3541 ccatctgcga cagggttacg gactggagga gcatccgtaa gtgtcatgac ccttccaacg
     3601 tagccattga atggctgagg gtagtcaact agggcttgcc aagcccccac ttcaaaagcc
     3661 gttaggcttt aagtggtggg gagttgacct tccctacaca caagcgccct tgttgcaacg
     3721 gctccgcact cgaggcctat agtgagtcgt attaatttct cccggacacc atcgaatggc
     3781 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
     3841 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
     3901 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
     3961 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
     4021 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
     4081 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
     4141 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
     4201 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     4261 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     4321 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     4381 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     4441 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     4501 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     4561 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     4621 ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc gaagacagct
     4681 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     4741 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     4801 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     4861 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4921 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg
     4981 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactaac
     5041 atgagaat
//
