LOCUS       sequence-411023-        5431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISCvi1 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5431)
  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 5431)
  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_ISCvi1_reRNA
FEATURES             Location/Qualifiers
     source          1..5431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..50
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             66..77
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1474..1491)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1644..1730
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1822..1849
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1868..1959
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2417..2872
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2504..2523)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2714..2735
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3428..4087)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4002..4021
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4088..4190)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4582..4599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4716..5261)
                     /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(4796..4815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5337..5356
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cagcttatca tcgatctgac agctagctca gtcctaggta taatgctagc cccgggaaaa
       61 gatctaaaga ggagaaaatg aacaaaggtg ttaagtttag ggcgtacccc aataaggaac
      121 aacaaaactt aataaaccga acactcggat gttgcaggct catctacaac aaaggtcttg
      181 ctatgcgtaa cgatgcctat tccaatggcg aaaagattgg ctataaccaa acatctgcta
      241 tgctcacaaa ccttaaaaag caggctgact ttgctttcct gaaggatgtt gattctattg
      301 cactccagca gtctttgagg gatttagacc gtggatttaa aaatttcttt gagaagcgtg
      361 caagacatcc gcagttcaaa agtaagcata ataaccacca gtcatacaga accatcaatc
      421 agggcgataa catccgcata atcgggaagt atattaagct cccgaagctc gggtatgtca
      481 aaatcaaaca gtccatggaa gtcgggcata tcaataatgt cacggtagag cgtactccta
      541 cgggtaaata ttttgtagtt ctgaatgttg actttgagcc agagtttcga ccaaatgcag
      601 gctgcatgat tggcattgat gtcggcataa aggagtttta ctccgacagc aacggcaatg
      661 tcgtcaataa ccctaagtat ttagagaagt caatgcgtaa actcgccaga gaacaacgca
      721 ggctttctcg taagcaaaaa ggctctaata atcgcaatag acagcgtgtt aaggtagcta
      781 aagtccatga aaagataact aatcagcgta acgatttcct tcaaaaacag tcaacgatac
      841 ttgttagcga aaaccaaacg atctgtatcg aagacctgaa tgtaaaagga atgattcgta
      901 atcataaact tgcacaacac atagcaagct gttcatggtc taagttcttt gcaatgcttg
      961 aatacaaggc tatatggtac ggaaatgaca taatcaaagt acctactatg tatccaagca
     1021 gtcagacatg cagttgttgt gggtataaga acccgtttgt aaagaatctc tcaatcagag
     1081 aatgggaatg tcctaattgt cacgcaaagc acgatagaga caccaacgca agcataacca
     1141 tcttaaataa agggctgtca atagcctgag agatctttga cagctagctc agtcctaggt
     1201 ataatagcaa agcacgatag agacaccaac gcaagcataa ccatcttaaa taaagggctg
     1261 tcaatagcct gacatatata aaataccgta gggcatacgg gaatttacgc ttgtggacac
     1321 tgtgtaagac gagccaacaa ggtaacttgt taggaacgca gtagtggttg aagcaagaat
     1381 cccacgactt tagtcgtgtg gagtgtcaag ctcggagatc atcattgcgt tttttaagct
     1441 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     1501 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     1561 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     1621 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1681 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     1741 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     1801 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     1861 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1921 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1981 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2041 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcag caaactatta actggcgaac
     2101 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2161 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2221 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2281 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2341 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2401 tactttagat tgatttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2461 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2521 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2581 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     2641 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2701 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     2761 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2821 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttaaaaggat
     2881 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3121 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3181 gcctacatac ctcgctctgc taatcctgtt accagtcagg catttgagaa gcacacggtc
     3241 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
     3301 cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc
     3361 gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt
     3421 aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     3481 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     3541 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     3601 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     3661 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     3721 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg
     3781 aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca
     3841 ccagctcacc gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa
     3901 gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg
     3961 ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct
     4021 caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt
     4081 tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     4141 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt
     4201 ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt
     4261 ctgcgaagtg atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg
     4321 ccaacttact gatttagtgt atgatggtgt ttttgaggtg ctccagtggc ttctgtttct
     4381 atcagctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc
     4441 ggacatcagc gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca
     4501 gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg
     4561 atacaggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg
     4621 gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact
     4681 taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac
     4741 aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga
     4801 taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc ctttcggttt
     4861 accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc
     4921 cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg
     4981 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc
     5041 actggcagca gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt
     5101 aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt
     5161 tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt
     5221 tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat cttattaatc
     5281 agataaaata tttctaggct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5341 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5401 cgtccggcgt agaggatctg ctcatgtttg a
//
