LOCUS       sequence-411098-        5528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISAeme8 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5528)
  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 5528)
  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_ISAeme8_reRNA
FEATURES             Location/Qualifiers
     source          1..5528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..50
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             66..77
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1243..1277
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1571..1588)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1741..1827
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1919..1946
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1965..2056
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2514..2969
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2601..2620)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2811..2832
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3525..4184)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4099..4118
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4185..4287)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4679..4696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4813..5358)
                     /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(4893..4912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5434..5453
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cagcttatca tcgatctgac agctagctca gtcctaggta taatgctagc cccgggaaaa
       61 gatctaaaga ggagaaagaa ttcatgactt tgagaagagc aacgaaagta cgcatttatc
      121 cgaccgacga acaagcggca ttcttgaatg cccagttcgg cgcggtgcgg ttcgcgtaca
      181 acaaggccct tcatattcaa cgacacatgt tcaagcgtca cgggatttca ctgaaaccga
      241 aacgcgacct gaaacccatg ctcgcagtgg cgaaaaaatc gcgcaaatac ggctggctaa
      301 aggagtacga ttcactcgcc ttgcagcaag cggtgattaa tctcgacaag gcattcgcca
      361 acttcttcaa tcccaagctc aaggcacgaa tgcctacctt caagagcaaa cggggcaggc
      421 aatccagcta tcaccccaac ggaaaggtgc tcacagcggc tatccagttg ccgaagatga
      481 cgcccatcag ggcggtcatt catcgagata ttatcggcgt ggtatccagc atcacagtca
      541 gccgtggccc gacagggaaa tactatgctt ccatcctttg cgaggatggc tgcgaggctc
      601 ccgccaagcc caccttcatc acggacgtga ctggctgtga tatggggctg tcccactacc
      661 tcattgagtc gagcggcaaa aagattgcca atccacgcca ccttatcaac gccagtcgca
      721 atctacgacg caagcaaaag gcactgtctc gcaagaaaaa gggcagcgcc aaccgtggca
      781 aggcaagacg gcaactggcc gccctgcacg aacgggtagc caatgcccgc gccgattttc
      841 agcacaaact ctctcacacg atggttgacg aaaaccaagc ggtcatcgtg gagacattga
      901 aatcagcgaa catgatgaaa aatcacaaac tggcccgcgc cattggcgat gctggttggc
      961 atggtttcat catgaagctg gagtacaagg cacaagcggc gggtcgccac ctaatcaaac
     1021 ttgatcagtg gtttgccagc tcgaaactgt gtagcgattg cggccacaag atgccggaca
     1081 tgccgttgca tcagcggcag tgggtatgtc cagcgtgtgg ggcagggcat gaccgcgaca
     1141 ttaacgcggc catgaatatc cagcaacaag gcatattgga attaaaggcg gcgggacacg
     1201 tcgtttctac ccatggaggc cagcgtggat cctgagagat ctttgacagc tagctcagtc
     1261 ctaggtataa tactagtgcc ggacatgccg ttgcatcagc ggcagtgggt atgtccagcg
     1321 tgtggggcag ggcatgaccg cgacattaac gcggccatga atatccagca acaaggcata
     1381 ttggaattaa aggcggcggg acacgtcgtt tctacccatg gaggccagcg ttaatctgcc
     1441 aattcggcag tagcggccta agaagtggga agcctcgccg cttgcggcgg ggagcagtca
     1501 cgctcggaga tcatcattgc gttttttaag cttagcttgg ctgttttggc ggatgagaga
     1561 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
     1621 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
     1681 gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat
     1741 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     1801 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     1861 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
     1921 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
     1981 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2041 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2101 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2161 agatcagttg ggtgcagcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2221 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2281 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2341 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2401 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2461 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc
     2521 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2581 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2641 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2701 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     2761 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2821 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     2881 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2941 ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata
     3001 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3061 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3301 tcctgttacc agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa
     3361 accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac
     3421 cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg
     3481 tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
     3541 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     3601 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     3661 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     3721 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     3781 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     3841 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     3901 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     3961 atacggaact ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     4021 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     4081 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     4141 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     4201 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     4261 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
     4321 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     4381 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg
     4441 atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc
     4501 tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt
     4561 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     4621 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     4681 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     4741 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     4801 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     4861 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
     4921 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     4981 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     5041 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     5101 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     5161 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     5221 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     5281 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     5341 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctaggctcat
     5401 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     5461 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc
     5521 atgtttga
//
