LOCUS       sequence-410910-        5581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISSen6 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5581)
  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 5581)
  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_ISSen6_reRNA
FEATURES             Location/Qualifiers
     source          1..5581
                     /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        1313..1347
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1631..1648)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1801..1887
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1979..2006
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2025..2116
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2574..3029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2661..2680)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2871..2892
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3585..4244)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4159..4178
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4245..4347)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4739..4756)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4873..5418)
                     /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(4953..4972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5494..5513
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgtttgacag cttatcatcg atctgacagc tagctcagtc ctaggtataa tgctagcccc
       61 gggaaaagat ctaaagagga gaaagaattc atgatgctaa tcctgaaagc ctacaaattc
      121 agactggaac caacgcctga acagtcgcag cgtttgcggc agttatgtgg ttgcgcccgt
      181 tttgtctgga atttaggtct tgcggagaca aagcgcatcc ttggttcagg cgaaaagtta
      241 ccttcggctt tcgagttgaa caggatgctt acagtgtgga aaaaaatgcc gggacacatc
      301 tttttacagg atgcttatac cgacaatctg caacaaaagt tgaaagacct gcatactgca
      361 tggaaacgtt gctttgataa aaagctcgca gctaaggccc cggtatggaa acgaaaaaat
      421 gagggcagag actcaatccg ttttgtgaac tttgagaaat attgccgcct tgaaaatcgc
      481 agagtgaagc taccgtcagg tcttgggtgg gtaaaattcc ggcaatctca acgtgtgaac
      541 ggtaaaatca aaaatgcgac aatcagtcag ttagcgggac agtggtatat ctcgtttcag
      601 gttgaagttg aaacggcaga acccaatcac acaagcacaa cgatagtcgg tctggacgca
      661 ggagtaacaa aactcgccac gctttcagac ggtacggtat accaaccagt caacagtttt
      721 aaagcaagtc agcgcaagct ggcaacactc cagcggcagt taagccgtaa agtcagattc
      781 agcagtaact ggcagaaaca aaagcgaaaa gtccagcatc tgcactcgca catagccaat
      841 atccggcgcg actaccttca taaagttacc agtgaaatca gcaaaaacca cgcgatgata
      901 gtcattgagg atttgaaggt tagtaacatg tcgaaatcgg caaaaggtac agcagaacag
      961 cacggacgaa acgtcaaggc caaatcaggt ctgaaccgtt cgatactgga tcagggctgg
     1021 tatgaaatgc gccgccagct tgagtacaaa cagctctggc gtggcggtca ggtgcttgcg
     1081 gttccgccag catatacaag ccagcgttgc gcgtgctgtg gtcataccgc gaaagaaaat
     1141 cgcctgtcac aaagtaaatt tgtgtgtcag gcatgcggat atacagcgaa cgctgatgta
     1201 aacggcgctc gtaacatttt agcggcgggg cacgccgttc ttgcctgtgg agggatggtg
     1261 cagtcaggcc gcccgtctga aacaggaacc agacgcggat cctgagagat ctttgacagc
     1321 tagctcagtc ctaggtataa tactagtgta aatttgtgtg tcaggcatgc ggatatacag
     1381 cgaacgctga tgtaaacggc gctcgtaaca ttttagcggc ggggcacgcc gttcttgcct
     1441 gtggagggat ggtgcagtca ggccgcccgt ctgaaacagg aaccagacgc tgagcaatca
     1501 gtcgtcaccc actaatggga attccccacc tgagtcacga agtgcgaagg tgggggagga
     1561 tgtcaagctc ggagatcatc attgcgtttt ttaagcttgg ctgttttggc ggatgagaga
     1621 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
     1681 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
     1741 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
     1801 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     1861 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     1921 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
     1981 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
     2041 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2101 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2161 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2221 agatcagttg ggtgcagcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2281 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2341 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2401 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2461 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2521 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc
     2581 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2641 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2701 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2761 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     2821 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2881 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     2941 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3001 ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata
     3061 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3121 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3181 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3241 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3301 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3361 tcctgttacc agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa
     3421 accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac
     3481 cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg
     3541 tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
     3601 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     3661 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     3721 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     3781 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     3841 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     3901 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     3961 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     4021 atacggaact ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     4081 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     4141 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     4201 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     4261 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     4321 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
     4381 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     4441 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg
     4501 atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc
     4561 tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt
     4621 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     4681 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     4741 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     4801 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     4861 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     4921 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     4981 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     5041 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     5101 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     5161 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     5221 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     5281 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     5341 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     5401 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctaggctcat
     5461 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5521 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc
     5581 a
//
