LOCUS       sequence-410872-        5598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISAs26 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5598)
  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 5598)
  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_ISAs26_reRNA
FEATURES             Location/Qualifiers
     source          1..5598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..43
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             59..70
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1317..1351
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1634..1651)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1804..1890
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1982..2009
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2028..2119
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2577..3032
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2664..2683)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2874..2895
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3588..4247)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4162..4181
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4248..4350)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4742..4759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4876..5421)
                     /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(4956..4975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5497..5516
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcatcgatct gacagctagc tcagtcctag gtataatgct agccccggga aaagatctaa
       61 agaggagaaa gaattcatgc aaattctgaa agcctacaaa ttccgactcg aaccgaccga
      121 acaacaagcg caacgcctgc gccagttggg cggttgtgct cgttatgtct ggaatcacgg
      181 gctggcagaa accaaacgca ttctggatgc tggcgaaaaa ctgccttctg cctttgagct
      241 gaacaggatg ctcaccagtt ggaagaaaga cccagaaatg gcctttctgg ccgaggcata
      301 taccgacaac ctccagcaga agctcaagga tttgcatggc gcgtggatgc gctgctttga
      361 taagaaactc gccgccaagg cacccgtatt caagaaaaaa agcgacaacc gcgacgctat
      421 ccggttcgtc aacttcgaca agtattgccg ccttgaaaag ggtcgcgtga agctgcctgc
      481 cgggctggcg tgggtcaagt tccgtcagtc ccagccagtt gaagggaaaa ttaaaaacgc
      541 gaccgtcagc caagcagccg ggcactggta tatctccttt caggtcgagc aggagacgct
      601 tgagccaatt cacccctccg gctcaatggt cgggctagat gcggggatcg ccaagctcgc
      661 aaccctgtca gacggcacgg tgtttgagcc cgtcaacagc ttcaaaacca atcagaccaa
      721 gctggcaagg ctccaacgcc agttgagcaa aaaggtaaag ttcagcagca actggaagaa
      781 acaaaaagcc aagatccagc ggctgcactc acatatcgcc aatacccgcc gcgattacct
      841 tcacaaggtc acaacgacta tcagcaaaaa ccacgcgatg atcgtcattg aggatttgaa
      901 ggttgccagc atgtcaaaat cagcagcggg cacgaccagc cagccggggc gcaacgtccg
      961 ggcaaaatca gggctgaatc gttcgatatt agatcagggc tggtttgagc tgcgccgcca
     1021 gctcgaatat aagcaggcat ggcgaggcgg tcaggtgttg gcgataaacc cggcctacac
     1081 cagccagcag tgcgcctgtt gtgggcatac tgcgaaggat aaccgccaga cgcaaagcca
     1141 gtttaaatgc gctgcgtgtg gttatgaggc gaacgccgat gtgaacggcg ctcgcaatat
     1201 tttagcggcg ggacacgccg ttttagcctg tggaggcgtg gtgcaatcag gccgcccgat
     1261 gaagcaggaa cccggtactg cgcgagcagc tatcacccag ggatcctgag agatctttga
     1321 cagctagctc agtcctaggt ataatactag tgccagttta aatgcgctgc gtgtggttat
     1381 gaggcgaacg ccgatgtgaa cggcgctcgc aatattttag cggcgggaca cgccgtttta
     1441 gcctgtggag gcgtggtgca atcaggccgc ccgatgaagc aggaacccgg tactgcgcga
     1501 gcagctatca cccagtaatg ggaatccccc tcctgtgacg cgaagcgcta aggtggggga
     1561 ggatgtcaag ctcggagatc atcattgcgt tttttaagct tggctgtttt ggcggatgag
     1621 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
     1681 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
     1741 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
     1801 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     1861 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
     1921 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     1981 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     2041 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2101 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2161 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2221 tgaagatcag ttgggtgcag caaactatta actggcgaac tacttactct agcttcccgg
     2281 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2341 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2401 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2461 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2521 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttacgc
     2581 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2641 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2701 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2761 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     2821 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2881 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     2941 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3001 gaattttaac aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg
     3061 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3121 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3181 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3241 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3301 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3361 taatcctgtt accagtcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     3421 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     3481 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     3541 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     3601 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     3661 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     3721 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     3781 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     3841 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     3901 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     3961 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     4021 gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     4081 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     4141 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     4201 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     4261 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     4321 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag
     4381 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
     4441 acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt
     4501 atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc
     4561 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag
     4621 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     4681 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     4741 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     4801 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     4861 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     4921 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4981 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     5041 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     5101 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     5161 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     5221 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     5281 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     5341 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     5401 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctaggct
     5461 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     5521 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg
     5581 ctcatgtttg acagctta
//
