LOCUS       sequence-411027-        5507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISClte2 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  AUTHORS   Xiang G, Li Y, Sun J, Huo Y, Cao S, Cao Y, Guo Y, Yang L, Cai Y, 
            Zhang YE, Wang H
  TITLE     Evolutionary mining and functional characterization of TnpB 
            nucleases identify efficient miniature genome editors.
  JOURNAL   Nat Biotechnol. 2023 Jun 29. doi: 10.1038/s41587-023-01857-x.
  PUBMED    37386294
REFERENCE   2  (bases 1 to 5507)
  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: "Nat 
            Biotechnol. 2023 Jun 29. doi: 10.1038/s41587-023-01857-x."
COMMENT     Sequence Label: pBad33_ISClte2_reRNA
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..54
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             70..81
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1241..1275
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1554..1571)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1724..1810
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1902..1929
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1948..2039
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2497..2952
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2584..2603)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2794..2815
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3508..4167)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4082..4101
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4168..4270)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4662..4679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4796..5341)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4876..4895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5417..5436
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttgacagctt atcatcgatc tgacagctag ctcagtccta ggtataatgc tagccccggg
       61 aaaagatcta aagaggagaa agaattcatg tgtcaactgt tggtggtgct ccactttcag
      121 caattaaaca atatattgaa aatcaaaaaa catcacaaag gaaatgatac tgtgcaatta
      181 accattaaaa tcaaattaat accaatgaaa gagcaaaaac tacatattca agatacttgt
      241 ttagaatata ttaaaactgt taatgatata gtatcactaa tggttgctga aaataaaagt
      301 cttaaactaa cttcaaaaga tataattgct aatcttccaa gtgctgttaa aaatcaagca
      361 attagagata gtaaaagtgt atattctaaa tataagaaaa ctaaaataca aaattttctt
      421 aaaaaaccta tgtgtatatg gaacaaccaa aactatagta ttggtgaaaa ttatattgct
      481 tttccagtaa tgattaatgg taaatctaaa aagattaaag ttaaagctat catagaacaa
      541 tatcaaaaag atttattaac caataaatta ggtactttaa gaataactaa aaaagccaat
      601 aaattaatgg ctcaaatttc tattgatgta gtagaaaaag aaaatgtaaa caataaagct
      661 atgggtatag atttgggact aaaaatacct gctgttgcag taacagaaga taaaaaagta
      721 aaattctttg gcaatggtag gcaaaacaaa tacattaaaa gaaaatatag atcattaaga
      781 aaaaagttag gtaaagctaa aaaaattaac aaaataaaat ctataaatga taaagaacaa
      841 agatatatga aagaccaaga ccacaaaata agtagaggaa tagttaattt tgctattcaa
      901 aataatattt ccgtaattag attagaaaaa ctaactaata tcagaaatac ggcaaaaaca
      961 aaccgtaaaa atgaaaagaa tttgcatact tggtcattct atcgtctagc acaatacatt
     1021 gaatataagg cacttttaga aggtattaaa attgaatatg ttaatccatg taatacatct
     1081 aaacaatgcc ctatttgtgg caaattaaat actgctaaag atagaaaata tcaatgttca
     1141 tgtggttata aagcacatag agatttgaat gcttgtatta atataattaa tgcaactgtt
     1201 gtagatggta atagtctaca agccggatcc tgagagatct ttgacagcta gctcagtcct
     1261 aggtataata ctagtgtggt tataaagcac atagagattt gaatgcttgt attaatataa
     1321 ttaatgcaac tgttgtagat ggtaatagtc tacaagccta agatactata ggtactgtct
     1381 taggaagggt actggcatac ccttagcttg aggtcatact ggactatcag aaatgaactg
     1441 tcctttaatc actcaagaat cccctagatt tatctatggg gagtgtcaag ctcggagatc
     1501 atcattgcgt tttttaagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     1561 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     1621 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     1681 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     1741 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1801 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     1861 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     1921 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     1981 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2041 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2101 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcag
     2161 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2221 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2281 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2341 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2401 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2461 agaccaagtt tactcatata tactttagat tgatttacgc gccctgtagc ggcgcattaa
     2521 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2581 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2641 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2701 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2761 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2821 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     2881 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2941 cgtttacaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3001 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3061 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3121 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3181 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3241 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtcagg
     3301 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     3361 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt
     3421 cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta
     3481 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtactg
     3541 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     3601 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     3661 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     3721 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     3781 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     3841 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     3901 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga actccggatg
     3961 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     4021 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     4081 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     4141 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     4201 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     4261 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
     4321 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
     4381 aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg
     4441 ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc
     4501 gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt
     4561 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
     4621 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
     4681 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     4741 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     4801 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     4861 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     4921 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     4981 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     5041 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5101 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     5161 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     5221 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     5281 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     5341 agaagatcat cttattaatc agataaaata tttctaggct catgagcccg aagtggcgag
     5401 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     5461 cggtgatgcc ggccacgatg cgtccggcgt agaggatctg ctcatgt
//
