LOCUS       sequence-411034-        5507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISBth16 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  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 5507)
  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_ISBth16_reRNA
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..43
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             59..70
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1543..1560)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1713..1799
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1891..1918
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1937..2028
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2486..2941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2573..2592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2783..2804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3497..4156)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4071..4090
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4157..4259)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4651..4668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4785..5330)
                     /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(4865..4884)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5406..5425
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcatcgatct gacagctagc tcagtcctag gtataatgct agccccggga aaagatctaa
       61 agaggagaaa gaattcatga caaagcagaa taaagcatat aaattccgtt tgtatccaac
      121 aaaagaccaa gcacatctta tgcgtaaaac cttcggttgt gtacgtttcg tgtataacag
      181 aatgttagct gaacggaaag aagcgtatga aaaacataaa gatgataagg atcaactaaa
      241 gaaacaaaag cttcccactc ctgcaaaata caaagcagaa tttgagtggt taaaagaagt
      301 ggattcatta gctttggcaa atgctcaact aaacttgcaa actgcctata aaaatttctt
      361 tcgtggtcaa aatgacttcc caacattcaa aagcaaaaaa gacagaaagt cttatacaac
      421 gaatgtagtg aacggaaata ttatattgct taacggtcat atcaaattgc caaaactgaa
      481 aatggtacgt atcaaacagc atagagaaat accacaaggc catataatca agtcatgtac
      541 gatttctatg acacctactg gtaaatacta tgtgtccatt ttgactgaat acgaaaaaga
      601 gatcgtacaa aaagaagtag aaacagttgt tggattagac tttgcgatgg atcaattata
      661 cgtcagttct gaggatgaga gagccaatta tcctaagttc tatcgtgaaa tgttagaccg
      721 attagcaaag gctcaacgag tattatcaag acgtacaaaa ggttcagggc gttggaataa
      781 acaacgtatc cgtgtagcta agctgcatga aaaagtagca aaccaacgta agaacttcct
      841 tcatcataag tctaaagagt tagctactca ttttgatgtt gtagctattg aagatctaaa
      901 tatgaaagga atgtcgcaag cacttcattt tggaaaaagt gtcgctgaca atggttgggg
      961 tatgttcact tcttttttag cttataaact acatgaacaa ggcaaacaac ttgtgaaaat
     1021 agacaaatgg tttccttcca caaaaacatg tagcagttgt ggaaatatga aaaatatgtc
     1081 attgtctgag cgagtctatt cttgtatatg cggtgtaaat ctcgatagag attataacgc
     1141 agctatcaat atcaaaaatg aagcaatacg cctattggcg ttagtaggat cctgagagat
     1201 ctttgacagc tagctcagtc ctaggtataa tagtctgagc gagtctattc ttgtatatgc
     1261 ggtgtaaatc tcgatagaga ttataacgca gctatcaata tcaaaaatga agcaatacgc
     1321 ctattggcgt tagtatagat agcaataaaa taacccttgg aacaagggag ttagctcggc
     1381 tgtcttagga cgaattcgtt agctaccaaa agtaacgact aaccgagaag cccccacttc
     1441 aaacagtccg ttaggacgtt aagtggcggg tagttcacgc tcggagatca tcattgcgtt
     1501 ttttaagctt ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc
     1561 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
     1621 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
     1681 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     1741 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     1801 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
     1861 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
     1921 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1981 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2041 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2101 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcagc aaactattaa
     2161 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2221 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2281 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2341 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2401 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2461 actcatatat actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2521 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2581 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2641 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2701 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2761 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2821 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2881 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     2941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3001 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3061 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3121 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3181 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtcaggc atttgagaag
     3301 cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg
     3361 gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc
     3421 cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat
     3481 aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     3541 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     3601 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
     3661 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     3721 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     3781 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     3841 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     3901 cccatatcac cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca
     3961 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     4021 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     4081 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     4141 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     4201 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     4261 cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat
     4321 ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg
     4381 gtgatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct
     4441 tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa
     4501 agcaccgccg gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg
     4561 agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca
     4621 gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     4681 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     4741 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     4801 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     4861 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
     4921 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
     4981 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
     5041 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
     5101 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
     5161 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
     5221 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
     5281 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
     5341 ttattaatca gataaaatat ttctaggctc atgagcccga agtggcgagc ccgatcttcc
     5401 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     5461 gccacgatgc gtccggcgta gaggatctgc tcatgtttga cagctta
//
