LOCUS       sequence-410935-        5543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISEc46 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5543)
  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 5543)
  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_ISEc46_reRNA
FEATURES             Location/Qualifiers
     source          1..5543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        12..46
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             62..73
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1263..1297
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1582..1599)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1752..1838
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1930..1957
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1976..2067
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2525..2980
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2612..2631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2822..2843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3536..4195)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4110..4129
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4196..4298)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4690..4707)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4824..5369)
                     /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(4904..4923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5445..5464
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttatcatcga tctgacagct agctcagtcc taggtataat gctagccccg ggaaaagatc
       61 taaagaggag aaagaattca tgttaagagc aacaaaagta cgcatctacc cgacaccaga
      121 acaggctgaa tttatcaacg cccagttcgg tgcagtgcgt tttgcgtaca acaaagctct
      181 gcatatcaag aaacattctt accagcggca ccgcgtaaat ttaaacccgc gtaaagacct
      241 caaaccactg cttgcagtgg cgaagaaatc ccgccggtat ggatggctta aagaatatga
      301 ttctattgcg ttgcagcagg cggtaattaa cctcgacgtg gctttttcta actttttcaa
      361 tccgaaactg aaagcaggta aacctacctt taaaagtaaa cacggcagac aatcgagcta
      421 tcattgtgtt ggcataaaag tccttgatgg tgcgataaaa ataccgaaaa tcttgccgat
      481 tgaagcgcgt ttacatcgtg aaattacggg aatgctgaaa agtatcacgt tgagccgcag
      541 cgcaaccggg aaatactacg cgtcgttact ttgtgacgac gggatggaag cgccggcaaa
      601 gcctgcactg atatcaaatg tgacagggct ggatatgggg ctggcgcatt ttgtcatcaa
      661 atcggacggc aataaaattg ccagtccccg ccacctaatc aatgcaagtc gcaaccttcg
      721 ccggaagcag aaagcactgt cccgtaagaa aaaagggaac gcaaacaggc gcaaagccag
      781 aatacaactt gcagccgttc acgaacgggt agccagtgcc cgtgccgatt tccagcacaa
      841 gttatctcgc gcaattgttg acgaaaacca agcggtaatt gtcgagacat tgaaaacggc
      901 aaatatgatg aaaaatcacc atctggctcg tgcgataggc gatgctggat ggtctggatt
      961 catcacaaaa ctggaataca aagcaacaga gaaaggtgtg catctggtga aactggatca
     1021 atggttcgcc agttcgaaaa cctgccactg ctgcggtcac aaaatgtcgg agatgccatt
     1081 acataagcgc atctggcagt gtccagagtg tggagttgaa catgaccgcg atatcaacgc
     1141 ggcgctcaac atcaaacgca agggcatact ggaattacaa gcggcgggac tcgtcgtttc
     1201 tgcccatgga ggccagcata aatccgtcac acagacggtt gcggctggat cctgagagat
     1261 ctttgacagc tagctcagtc ctaggtataa tactagtgag atgccattac ataagcgcat
     1321 ctggcagtgt ccagagtgtg gagttgaaca tgaccgcgat atcaacgcgg cgctcaacat
     1381 caaacgcaag ggcatactgg aattacaagc ggcgggactc gtcgtttctg cccatggagg
     1441 ccagcataaa tccgtcacac agacggttgc ggcttaagaa gtgggaagcc tcgcccgaca
     1501 gggcggggag cagtcacgct cggagatcat cattgcgttt tttaagcttg gctgttttgg
     1561 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     1621 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     1681 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     1741 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1801 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     1861 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     1921 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     1981 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2041 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2101 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2161 aaagatgctg aagatcagtt gggtgcagca aactattaac tggcgaacta cttactctag
     2221 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2281 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2341 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2401 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2461 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2521 atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2581 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2641 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2701 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     2761 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2821 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
     2881 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2941 tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat
     3001 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3061 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3241 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3301 cgctctgcta atcctgttac cagtcaggca tttgagaagc acacggtcac actgcttccg
     3361 gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct
     3421 gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac
     3481 ttattcaggc gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg
     3541 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     3601 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     3661 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     3721 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
     3781 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     3841 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     3901 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     3961 ctttcattgc catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa
     4021 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     4081 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     4141 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     4201 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     4261 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt
     4321 tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat
     4381 cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga
     4441 tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc
     4501 tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc
     4561 tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
     4621 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
     4681 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
     4741 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
     4801 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
     4861 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4921 ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt
     4981 ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt
     5041 tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc
     5101 cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc
     5161 cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg
     5221 aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg
     5281 tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag
     5341 agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt
     5401 tctaggctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     5461 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     5521 aggatctgct catgtttgac agc
//
