LOCUS       sequence-410905-        5489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISYmu1 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5489)
  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 5489)
  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_ISYmu1_reRNA
FEATURES             Location/Qualifiers
     source          1..5489
                     /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)"
     primer_bind     complement(1536..1553)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1706..1792
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1884..1911
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1930..2021
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2479..2934
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2566..2585)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2776..2797
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3490..4149)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4064..4083
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4150..4252)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4644..4661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4778..5323)
                     /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(4858..4877)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5399..5418
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttgacagctt atcatcgatc tgacagctag ctcagtccta ggtataatgc tagccccggg
       61 aaaagatcta aagaggagaa aatgctgcag cataaagcct atgaataccg tatctatcca
      121 gataagaagc aggaaacact gattgccaag acaataggca gttcaaggta tgtatacaac
      181 catttcctgg aactttggaa caaggaatat gaggagaccg gaaaaggtct gacttattac
      241 gcatgctcca aacttctgac aaagcttaag agggacccgg aaacagtatg gctttgtgaa
      301 gtggataagt tctcgctcca gaactcgctt cgtaacctgt cggacgcctt ctcccgcttt
      361 ttcaaaggac agaacgagca tccccagttc aagagcaaga agagcccgag gcaaagctac
      421 acgacacaat atacgaacaa caacatcgca gtttccggaa attgtctgaa gctgcctaag
      481 ctgggccttg tcaagtttgc agacagtagg gagatgaaag gccgtatact gaatgccaca
      541 gtacgaagga aatcgagcgg gaaattcttc gtatcgatcc tatgcgaaga ggagatctgt
      601 gaattgccaa agactgactc atctgtcgga attgacctcg gaatcattga ctttgcggtt
      661 atgtcagacg gaagcaggca tgacaacaat cattttacca ggcaaatgga agaaaggctt
      721 agacgggagc agcgaaagct tgcaaggcgt gcacttgctg cggagaaaag gggcatttcc
      781 ctttctgaag ccaggaacta tcagaagcaa aggcggaagg tagcgagact ttatgaaaag
      841 gttgcaaacc agcgaaaaga atatcttaac aaactcagta cagagatagt caaaaaccac
      901 gatatcatct gtatcgagga ccttaacgtt aagggcatga tgcgcaatca taaactggca
      961 aaaagcatct ctgatgtatc atggacgagc cttgtatcga aactgcagta caaggcttcc
     1021 tggtatggaa aagaagtcat caggataagc agatggtttc cgtcaagtca gatatgctca
     1081 gagtgcggcc acaaggacag gaagaagcct ctccatgtaa gggagtggac ctgtcctgtt
     1141 tgtcatgccc accatgaccg tgacgtcaat gcagcaagaa atatactggc tgagggactc
     1201 aggataagag ctctgactcc ggggtcttga gagatctttg acagctagct cagtcctagg
     1261 tataatagtc aatgcagcaa gaaatatact ggctgaggga ctcaggataa gagctctgac
     1321 tccggggtct tagcaataca gaagcaaaca ggaaccgcag gaattgcggg ggtagcttgg
     1381 taaacaagag aaacctctgc cggcaaagaa ataagccggt aagtatgctc tgttcccaag
     1441 aatctcgtga ctttagtcat gagagtttca agctcggaga tcatcattgc gttttttaag
     1501 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     1561 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     1621 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     1681 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     1741 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     1801 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     1861 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     1921 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1981 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2041 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2101 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc agcaaactat taactggcga
     2161 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2221 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2281 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2341 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2401 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2461 tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2521 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2581 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2641 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     2701 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2761 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2821 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2881 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg
     2941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     3001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3061 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3181 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3241 ccgcctacat acctcgctct gctaatcctg ttaccagtca ggcatttgag aagcacacgg
     3301 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     3361 aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc tgccattcat
     3421 ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc aataactgcc
     3481 ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     3541 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     3601 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     3661 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     3721 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     3781 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     3841 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     3901 caccagctca ccgtctttca ttgccatacg gaactccgga tgagcattca tcaggcgggc
     3961 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     4021 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc
     4081 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     4141 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     4201 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     4261 ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta
     4321 ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc
     4381 tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt
     4441 ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg
     4501 ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt
     4561 cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg
     4621 tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
     4681 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
     4741 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
     4801 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
     4861 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
     4921 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
     4981 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
     5041 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
     5101 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
     5161 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
     5221 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
     5281 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
     5341 tcagataaaa tatttctagg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     5401 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     5461 tgcgtccggc gtagaggatc tgctcatgt
//
