LOCUS       sequence-410889-        5652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of IS605 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5652)
  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 5652)
  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_IS605_reRNA
FEATURES             Location/Qualifiers
     source          1..5652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..56
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             72..83
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1701..1718)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1871..1957
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2049..2076
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2095..2186
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2644..3099
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2731..2750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2941..2962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3655..4314)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4229..4248
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4315..4417)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4809..4826)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4943..5488)
                     /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(5023..5042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5564..5583
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga tctgacagct agctcagtcc taggtataat gctagccccg
       61 ggaaaagatc taaagaggag aaagaattca tgcttaacgc tatcaagttt agaatttatc
      121 ctaacgctca acaaaaagag cttatttcta aacattttgg ctgttctagg gtcgtgtata
      181 actacttttt agattaccga caaaagcaat acgcaaaagg cattaaagaa acttacttca
      241 ccatgcaaaa agtcttaacc caaatcaagc accaagaaaa ataccattac ctcaatgaat
      301 gcaattctca aagcttgcaa atggcgttaa gacagcttgt gagtgcttat gataatttct
      361 ttagcaaaag agcgagatac cctaaattca aatctaaaaa aaacgctaaa caatcttttg
      421 caatccccca aaacatagaa atcaaaacag agactcaaac catcgctctc cctaaattca
      481 aagagggcat taaggctaaa ttacacagag aattgcctaa agatagcgtt atcaaacagg
      541 cttttatttc ttgcatagcc gatcaatatt tttgttctat atcctatgaa accaaagagc
      601 ctatccctaa acctaccatc attaaaaaag cggtaggttt agacatgggc ttaagaacgc
      661 tcattgttac aagcgataaa atagaatacc cacacatccg tttttatcaa aaattagaaa
      721 agaaactcac taaagcgcaa aggaggttaa gtaaaaaagt aaaaggctcc aacaacagga
      781 aaaaacaagc taaaaaggta gctagattgc atctagcttg ttcaaacact agagatgact
      841 acttgcataa aatcagtaat gagataacca atcaatacga tttgataggg gtagaaactt
      901 tgaatgttaa ggggcttatg agaacctatc attctaaaag ccttgctaat gcgagttggg
      961 ggaaattcct tactatgcta aaatacaaag cccaaagaaa agctaaaacc ctattaggca
     1021 tagacagatt tttccctagc tctcaattgt gttcttattg tgggttcaat acaggcaaaa
     1081 aacatgaaaa catcactaaa ttcacttgtc ctcattgcaa tatcacacac cacagagatt
     1141 acaatgcgag cgtcaatatt agaaactacg ctttaggcat gctagatgac aggcataaaa
     1201 taaagataga taaaagtagg gtagggatta tccgaactga ttacgctcat tacactgatg
     1261 agcgcatcaa agcttgtgga gcttcctcta atggggttat ttctaaatat ggcaacatat
     1321 tggatctagc tagttatgga gcgatgaagc aagaaaaagc ccaatcgctt ggatcctgag
     1381 agatctttga cagctagctc agtcctaggt ataatagctt taggcatgct agatgacagg
     1441 cataaaataa agatagataa aagtagggta gggattatcc gaactgatta cgctcattac
     1501 actgatgagc gcatcaaagc ttgtggagct tcctctaatg gggttatttc taaatatggc
     1561 aacatattgg atctagctag ttatggagcg atgaagcaag aaaaagccca atcgctttag
     1621 cggttgggta attcacgctc ggagatcatc attgcgtttt ttaagcttgg ctgttttggc
     1681 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     1741 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     1801 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     1861 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1921 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     1981 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     2041 aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt
     2101 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2161 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2221 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2281 aagatgctga agatcagttg ggtgcagcaa actattaact ggcgaactac ttactctagc
     2341 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2401 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2461 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2521 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2581 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2641 tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2701 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2761 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2821 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2881 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2941 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     3001 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3061 ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc
     3121 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3181 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3241 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3301 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3361 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3421 gctctgctaa tcctgttacc agtcaggcat ttgagaagca cacggtcaca ctgcttccgg
     3481 tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg
     3541 aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact
     3601 tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc
     3661 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     3721 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     3781 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     3841 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     3901 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     3961 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     4021 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     4081 tttcattgcc atacggaact ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     4141 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     4201 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     4261 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     4321 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     4381 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt
     4441 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
     4501 ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat
     4561 ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct
     4621 cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct
     4681 agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca
     4741 tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat
     4801 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
     4861 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
     4921 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
     4981 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5041 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
     5101 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
     5161 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
     5221 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
     5281 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
     5341 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
     5401 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
     5461 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt
     5521 ctaggctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     5581 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     5641 ggatctgctc at
//
