LOCUS       sequence-410924-        5465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISMma22 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5465)
  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 5465)
  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_ISMma22_reRNA
FEATURES             Location/Qualifiers
     source          1..5465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..57
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             73..84
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1515..1532)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1685..1771
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1863..1890
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1909..2000
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2458..2913
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2545..2564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2755..2776
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3469..4128)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4043..4062
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4129..4231)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4623..4640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4757..5302)
                     /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(4837..4856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5378..5397
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgtttgacag cttatcatcg atctgacagc tagctcagtc ctaggtataa tgctagcccc
       61 gggaaaagat ctaaagagga gaaagatttc atgttaaaag tgcttcggta tagagtctat
      121 cctacaaagg ctcaaaaaac aactatgaat aatactcttg agctttgtag gtggacatat
      181 aacgagacac ttgcactcag gaaagatgca tgggagtcag agcaaaagaa tatagggtac
      241 tatgattcta agaaaatgat ccctgaatgg aaaaaagaaa aacctgaatt gaaaacagtt
      301 cattcacagg tattacagga agtaatcaag agagtagatc ttgcctttca agcctttttc
      361 cgtagagtta aaacaggaga gaaggcaggc tatcctagat tcagaggata cggaaggtat
      421 gatagtttta cctatactca atcaggcttt tctctggatt ctgataggtt aacactatca
      481 aaaataggtg atattaaaat caaattacat agaccagtag aaggtgaaat taaaagactc
      541 aatgttcgta agatgcctac aggaaaatgg tttgtttctt ttctggttga aactgatact
      601 cctttggaat tacaaaaaac aggattatca gtaggtgtag acgtaggtat caagagcttc
      661 ttaacgcttt ctgatggtaa ttatgtacct aatcctagat tctttgttac agaagaaaag
      721 tttctggcaa aagtccaaag aaaactctcc aaacttgaaa aaggaacacc tgaaagagca
      781 aaagccttga aagctgttca acgtgttcac gaaagaatct caaacaagag agaggatttt
      841 gtacagaaag taagtcttaa tctggtcagg aattatgacc tgattacttt tgaagatttg
      901 aacattaaag gtatgattaa aaaccacaaa ctagcaaaac acatagctga tgttgcatgg
      961 aataagcttg taactatcac gtcttacaag gcagaatggg ctggtaaacg tgtagagctt
     1021 gtcaatccat gtaatacatc acaaatgtgc tctggatgcg gtgaaatagt taaaaaagaa
     1081 ctatcagaga gaatacacag ttgtccttac tgtggtctta ctcttgatag agatcacaat
     1141 gccgctatta acattatgag attgggatta caatctctcc agaactctgg tagatgcccg
     1201 tcacttcagg gatcctgaga gatctttgac agctagctca gtcctaggta taatagtaat
     1261 acatcacaaa tgtgctctgg atgcggtgaa atagttaaaa aagaactatc agagagaata
     1321 cacagttgtc cttactgtgg tcttactctt gatagagatc acaatgccgc tattaacatt
     1381 atgagattgg gattacaatc tctccagaat tctggtagat gcccgtcact tcagtgatgg
     1441 gagtagtcac gctcggagat catcattgcg ttttttaagc ttggctgttt tggcggatga
     1501 gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag
     1561 aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg
     1621 aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
     1681 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1741 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
     1801 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
     1861 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
     1921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1981 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2041 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2101 ctgaagatca gttgggtgca gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2161 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2221 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2281 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2341 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2401 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttacg
     2461 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2521 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2581 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2641 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     2701 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2761 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     2821 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2881 cgaattttaa caaaatatta acgtttacaa tttaaaagga tctaggtgaa gatccttttt
     2941 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3001 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3061 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3241 ctaatcctgt taccagtcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     3301 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     3361 cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca
     3421 ggcgtagcaa ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc
     3481 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     3541 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     3601 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     3661 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     3721 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     3781 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     3841 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     3901 tgccatacgg aactccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     3961 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     4021 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     4081 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     4141 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     4201 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca
     4261 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     4321 cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac tgatttagtg
     4381 tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctcctgtt
     4441 cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctagcgga
     4501 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     4561 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     4621 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     4681 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     4741 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     4801 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4861 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     4921 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     4981 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     5041 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     5101 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     5161 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     5221 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     5281 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctaggc
     5341 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     5401 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct
     5461 gctca
//
