LOCUS       sequence-410936-        5587 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISEc41 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5587)
  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 5587)
  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_ISEc41_reRNA
FEATURES             Location/Qualifiers
     source          1..5587
                     /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)"
     promoter        1316..1350
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1637..1654)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1807..1893
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1985..2012
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2031..2122
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2580..3035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2667..2686)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2877..2898
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3591..4250)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4165..4184
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4251..4353)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4745..4762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4879..5424)
                     /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(4959..4978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5500..5519
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgtttgacag cttatcatcg atctgacagc tagctcagtc ctaggtataa tgctagcccc
       61 gggaaaagat ctaaagagga gaaagaattc gaattcatgt taatccagaa agcatataaa
      121 tacaggctct accccactga ccagcaggcg caaaggttgc gtcagttgtg cggttgcgct
      181 cgctttgtct ggaactatgc actgaatgaa acactgagca ttcatgacgc gggcggaaaa
      241 attccgtcag ctttcgacct gaataaacga cttacgggct ggaaaaaact ccctgagctg
      301 gcctttcttt ctgagggtta cactgacaac ctccagcaga aactgaaaga cctgcggagc
      361 gcatgggatc gctgttttga taaatccctg acggctgaaa agcccgtttt caaaaagaaa
      421 accaaaggct gcgactctat ccggttcgtc aatttcagca aatattgtgg gctggattat
      481 ggccgtgtga aactcccgtc aggactgggc tgggttaaat ttcgtcagtc ccggaagatt
      541 gaaggcgtca ttaaaaattg caccatcagc cagcatgccg gacactggta tgtgtctttt
      601 caggtcgaat tagctgttac cgatccgatt cacgcatcga ccagcgcaat aggcctggat
      661 gcaggtataa cgaagctggc aacactttca gacggcacgg tctttgaacc ggtaaacagc
      721 ttaaaaaaaa atcaggataa gctggctcgc ttacagcgcc gtctggcccg gatggttaaa
      781 ttctccgcca actggaaaaa gcagaaagcg aagatttccc gtttccactc tcatatcgca
      841 aacatccgcc gcgattacct tcataaaacc acgacgacga tcagcaaaaa ccacgcgatg
      901 atagtcattg aggatttaaa ggtttcgaac atgtcaaaat cagcagcggg tacggtagac
      961 cagccggggc gcaacgtcgc ggcaaaatca ggtcttaacc gggcgatact ggatcagggc
     1021 tgggcagaaa tgcgccgcca gctcgaatat aagcaggcat ggcgcggcgg tgatgtactg
     1081 gcgattaatc cggcgtacac cagtcagaaa tgcgcatgtt gtgggcatac atcgaaaaat
     1141 aaccgcagga cgcaggcgag ctttatctgt accgcgtgcg gctatacggc aaacgccgat
     1201 gtaaacggcg ctcgcaatat attaacagca ggttttgaag tcatggcggc aggacaaata
     1261 ttgccgtcag tccgtaaagg acgcgcccgt aaggcggcgg gatcctgaga gatctttgac
     1321 agctagctca gtcctaggta taatactagt ctatacggca aacgccgatg taaacggcgc
     1381 tcgcaatata ttaacagcag gttttgaagt catggcggca ggacaaatat tgccgtcagt
     1441 ccgtaaagga cgcgcccgta aggcggcgta agccgtcgag cgaggaaccg gggaatgcgg
     1501 caacgtattg ctaccctgac aagggaatcc ccctcctgag tcgctttgcg cgaaggtggg
     1561 ggaggatgtc aagctcggag atcatcattg cgttttttaa gcttggctgt tttggcggat
     1621 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     1681 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     1741 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     1801 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1861 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     1921 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     1981 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     2041 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2101 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2161 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2221 tgctgaagat cagttgggtg cagcaaacta ttaactggcg aactacttac tctagcttcc
     2281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     2581 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2641 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2701 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2761 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     2821 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2881 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     2941 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3001 cgcgaatttt aacaaaatat taacgtttac aatttaaaag gatctaggtg aagatccttt
     3061 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3361 tgctaatcct gttaccagtc aggcatttga gaagcacacg gtcacactgc ttccggtagt
     3421 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
     3481 gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt atcacttatt
     3541 caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa ttacgccccg
     3601 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca
     3661 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     3721 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa
     3781 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac
     3841 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt
     3901 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc
     3961 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc
     4021 attgccatac ggaactccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc
     4081 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga
     4141 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga
     4201 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc
     4261 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta
     4321 tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc
     4381 cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc
     4441 gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt actgatttag
     4501 tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct gtccctcctg
     4561 ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc agcgctagcg
     4621 gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt gcttcatgtg
     4681 gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg atatattccg
     4741 cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt
     4801 acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg gaagtgagag
     4861 ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc acgaaatctg
     4921 acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4981 tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct
     5041 gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct
     5101 ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta
     5161 actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg
     5221 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg
     5281 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct
     5341 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt
     5401 acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag
     5461 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     5521 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     5581 ctgctca
//
