LOCUS       sequence-410874-        5459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISClsp11 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5459)
  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 5459)
  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_ISClsp11_reRNA
FEATURES             Location/Qualifiers
     source          1..5459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..37
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             53..64
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1489..1506)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1659..1745
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1837..1864
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1883..1974
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2432..2887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2519..2538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2729..2750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3443..4102)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4017..4036
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4103..4205)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4597..4614)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4731..5276)
                     /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(4811..4830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5352..5371
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atctgacagc tagctcagtc ctaggtataa tgctagcccc gggaaaagat ctaaagagga
       61 gaaaatgaac gtagtatacc ggtttcggat ctatcccaat aaaatacagc aggaactgtt
      121 tgcaaaaaca tttggatgtg tcaggttcgt atataaccgg atgctgtcag agaagaagga
      181 atgctatgaa aagaccggaa aaaatctaaa ggtcactccg gcaaaatata aggcagaatt
      241 tccatggcta aaagaagtgg acagccttgc tctgtgcaat gcccagctgc acctacagat
      301 tgcatataaa aactttttcc gggatccatc gtttggtttt ccgagattca agtcaaaaaa
      361 gaatcctgtt aaaagctata ccacaaacag tgtcaatgga aatatcctgt taaaagatgg
      421 gaagctgaag ctccccaaaa ctggctggat ccggatccgg cagcatcgta agatccggga
      481 agatgcctgc ctgaaaggcg caaccgtctg tatggaagct gatgggcggt attatgtttc
      541 tcttctgtat ttttgtgaag aaaagacggt ggaaacaagg aacataaggc aggccgtcgg
      601 actcgatttt tccatgaaag aactatatgt agattccaat ggaaaacatg caggttatcc
      661 gcactatttc agaaacagcc aggaaaaact ggcgaaagcg cagagaaaac tgagtcattg
      721 ccggaaagaa agcaatcgtt acaaaaaaca gcagaaaaaa gtggcgcgga tccatacaca
      781 tatcgctcac cagagaaaag attacttgca taaagaatca agaaagataa ccaatttcta
      841 tgatatcgta tgtatcgaag atctggatct gaaaacgatg agcggggaac atcattttgg
      901 aaaaagtgtc catgataacg gctggagaat gttcacagat tttttgcagt ataaactgga
      961 acgggaaggg aagaaacttg tccgcatcga ccgctggtat cccagttcca gaacatgcag
     1021 ctgttgtgga aaaataaaac atgatctgaa actggaagag cgggtatatc tttgtaactg
     1081 cggaaaccgg atggaccgtg atgaaaatgc ggcggtcaat atctgccggg aaggcctgag
     1141 aatatccggg atcatattag aaaaaagcga aaaagcatca tgagagatct ttgacagcta
     1201 gctcagtcct aggtataata gaaaatgcgg cggtcaatat ctgccgggaa ggcctgagaa
     1261 tatccgggat catattagaa aaaagcgaaa aagcatcata aaaagatatc agtccaaaag
     1321 aaccgtggga cacacgggga tcgctcgtga atgctttgcc caacggggca atcgaacgag
     1381 aagcccccac ttcaagcgtg agctaagtgg tgggagtatg tcacgctcgg agatcatcat
     1441 tgcgtttttt aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     1501 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     1561 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     1621 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     1681 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1741 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     1801 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     1861 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     1921 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1981 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2041 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcagcaaac
     2101 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2161 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2221 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2281 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2341 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2401 aagtttactc atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg
     2461 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2521 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2581 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2641 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2701 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2761 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2821 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     2881 acaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2941 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3001 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3061 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3121 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3181 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tcaggcattt
     3241 gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca
     3301 taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat
     3361 ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc
     3421 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta
     3481 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     3541 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     3601 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     3661 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     3721 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     3781 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     3841 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc ggatgagcat
     3901 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     3961 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     4021 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     4081 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     4141 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     4201 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac
     4261 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg
     4321 cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca
     4381 gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac
     4441 ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca
     4501 ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg
     4561 tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc
     4621 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     4681 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     4741 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
     4801 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
     4861 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
     4921 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
     4981 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
     5041 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga
     5101 agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa
     5161 gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca
     5221 aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag
     5281 atcatcttat taatcagata aaatatttct aggctcatga gcccgaagtg gcgagcccga
     5341 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     5401 atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttatcatcg
//
