LOCUS       sequence-410875-        5552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISRor9 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5552)
  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 5552)
  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_ISRor9_reRNA
FEATURES             Location/Qualifiers
     source          1..5552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..41
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             57..68
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1282..1316
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1586..1603)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1756..1842
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1934..1961
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1980..2071
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2529..2984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2616..2635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2826..2847
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3540..4199)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4114..4133
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4200..4302)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4694..4711)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4828..5373)
                     /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(4908..4927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5449..5468
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatctga cagctagctc agtcctaggt ataatgctag ccccgggaaa agatctaaag
       61 aggagaaaga attcgaattc atgcatagag cgacaaaagt acgcatttac ccaacgcaag
      121 aacaaactga attgctggaa aagcaattcg gcgcggtgcg ttttgcgtac aataaagccc
      181 tgtatattcg ctctcactgg tacaaaaaac atggcgtatc gctgaataat gcaaagcata
      241 ttcagcctct gctacccatt gccaaaaagt cccgccgcta tagctggctc aaggcagcag
      301 actccaacgc attgcaggta tctttgcaga atctggatcg tgcattccaa aacttctttg
      361 agaaacgggc gaaatatccg aagttcaaac gcaagcacgg accgcaatcc tcctatcatt
      421 gcagtagcct gaaaatcact gccgacacaa tcaagatccc gaagatttcc cctatccggg
      481 cgaacattca ccgggagatt gtcggcaagg tgaaaagcat caccatcagc cgcgcaccga
      541 ctggcaaata ctacgcctca atccttgccg atgatggaca ggaagcgcca gcgaagccga
      601 cctacataac ccaactgaca ggctatgaca ttggcctgaa agattatctg atacgcgatg
      661 acggcaaaaa aacgcctaac ccgcgtcact tggtcaatgc ctatcgccgc ttacgtcgcg
      721 atagcaaggc gctatcccgt aaacagaagg gaagcgcgaa tcgtcgcaag gcacgtattc
      781 gcctagcagc acaacacgat cgggtagcca atgcccgtaa cgattttcag cacaaacttt
      841 ccagaaatat tatcgtcgaa aatcaagcgg tgattgtgga aacactgaaa acggcaaaca
      901 tgatgaaaaa caaacgcctt gcccgtcata tcggggatgc cgcgtggtct ggtttcgtgg
      961 cgaaactggc ctataaagcg gaggcgtccg gcgtccacct ggtcaaactg gatcagtggt
     1021 tcgccagttc aaaaacatgc cattgttgcg gtcacaagat ggcagaaatg ccactgaaag
     1081 tacgtgaatg gcaatgccct gtctgcctga ctcaccataa tcgcgatatc aatgcggcga
     1141 tgaacatcag gctgaagggc ataacagaac taatggcggc gggtcacgtc gtcaaagccc
     1201 acggaggcct gcgtaagtcc gttataacaa cggttgcggc ctatgaagtg ggaagccccg
     1261 cccttggatc ctgagagatc tttgacagct agctcagtcc taggtataat actagtgtac
     1321 gtgaatggca atgccctgtc tgcctgactc accataatcg cgatatcaat gcggcgatga
     1381 acatcaggct gaagggcata acagaactaa tggcggcggg tcacgtcgtc aaagcccacg
     1441 gaggcctgcg taagtccgtt ataacaacgg ttgcggccta tgaagtggga agccccgccc
     1501 tttagggcgg ggagcagtca cgctcggaga tcatcattgc gttttttaag cttggctgtt
     1561 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
     1621 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
     1681 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
     1741 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1801 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     1861 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     1921 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     1981 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2041 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2101 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2161 agtaaaagat gctgaagatc agttgggtgc agcaaactat taactggcga actacttact
     2221 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     2281 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     2341 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     2401 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2461 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2521 attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2581 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2641 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2701 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     2761 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2821 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     2881 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2941 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg atctaggtga
     3001 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3061 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3121 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3181 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3241 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3301 acctcgctct gctaatcctg ttaccagtca ggcatttgag aagcacacgg tcacactgct
     3361 tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt aacgaccctg
     3421 ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat ccgcttatta
     3481 tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc ttaaaaaaat
     3541 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     3601 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     3661 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     3721 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
     3781 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     3841 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     3901 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     3961 ccgtctttca ttgccatacg gaactccgga tgagcattca tcaggcgggc aagaatgtga
     4021 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     4081 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     4141 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     4201 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     4261 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa
     4321 aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag
     4381 tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc tgccaactta
     4441 ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt ctatcagctg
     4501 tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg ccggacatca
     4561 gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg
     4621 cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga
     4681 tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg
     4741 aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg
     4801 aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca
     4861 cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc
     4921 gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt
     4981 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg
     5041 cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct
     5101 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag
     5161 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa
     5221 actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag
     5281 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag
     5341 caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa
     5401 tatttctagg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     5461 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     5521 gtagaggatc tgctcatgtt tgacagctta tc
//
