LOCUS       sequence-411035-        5482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISBce3 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5482)
  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 5482)
  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_ISBce3_reRNA
FEATURES             Location/Qualifiers
     source          1..5482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..52
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             68..79
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1179..1213
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1527..1544)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1697..1783
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1875..1902
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1921..2012
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2470..2925
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2557..2576)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2767..2788
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3481..4140)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4055..4074
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4141..4243)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4635..4652)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4769..5314)
                     /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(4849..4868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5390..5409
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gacagcttat catcgatctg acagctagct cagtcctagg tataatgcta gccccgggaa
       61 aagatctaaa gaggagaaag aattcatgtc acaaacaatc acagtcaaag ttaaattgct
      121 tccaacaaaa gaacaaattc agttattgga acaaagtagc tatgagtata tcaaagtcat
      181 taatacactt gtatctgaaa tggctgaagc gaagaagagc actaagaaaa gtacaaaaga
      241 tgttgaagcg aatcttccaa gcgcagtgaa gaaccaagca attaaagacg ctaaaagtgt
      301 gttttctaca aaggtgaaaa aaagtaaata caaagttatt ccgattctaa agagacctgt
      361 ttgtgtatgg aacaatcaaa actattcgtt tgacttcacg cacatttcag ttccactcat
      421 ggtaaatgga aaatcaactc gcttaaaggt tcgtgcttta ttaactgata aatataatcg
      481 caattttaat ttgttaacac acaagttagg tacacttcgt atcacgaaga aatctaataa
      541 gtggattgct caaatatctg tcacaattcc tacaaatgaa aaaacaggaa cgaagatttt
      601 aggggtagat ttaggtctta aagtccctgc ggtagccatc acagacgatg agaaagttcg
      661 tttcttcggc aatggtagac aaaacaaata tatgaaacgt aagtttcgta gtgttcgtaa
      721 gaagttaggt gaaaataaga aattaaatgc cattcgacaa cttgatgata aagaacaaag
      781 atggatgcaa gaccaagacc acaaagtaag tcgtgaaatc attgactttg caactgataa
      841 taacatttct gttattcgtt tagaacaact aacgaatatt agacagacgg caagaacaag
      901 ccgtaaaaac gagaaaaatc tacacacttg gtcattctat cgtttagcac aattcattga
      961 atacaaagcg aatttagttg gtattaaggt agaatatgtg aaccctgctt atacaagtca
     1021 aacatgtccg aaatgttctg aaaagaacaa agcaccagac agaaaataca agtgccgatg
     1081 tggatttgag aaacatcgtg atattgttgg ggcgatgaat attcgctacg caactgtgat
     1141 tgatggtaac agtcaatcag ccggatcctg agagatcttt gacagctagc tcagtcctag
     1201 gtataatact agtgtgccga tgtggatttg agaaacatcg tgatattgtt ggggcgatga
     1261 atattcgcta cgcaactgtg attgatggta acagtcaatc agcctaagat gctatacgca
     1321 ctgtcttagg aggggcaatg agatgccctc atcttgaagt ctgttcaaaa cagaaatgga
     1381 ctgcgaacgt ttagtcattc aagaatccca cccgtcaatc cgtaaggatt agggcttgcg
     1441 actttagtcg tgggagtctc aagctcggag atcatcattg cgttttttaa gcttggctgt
     1501 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1561 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1621 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1681 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1741 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     1801 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     1861 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     1921 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1981 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2041 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2101 aagtaaaaga tgctgaagat cagttgggtg cagcaaacta ttaactggcg aactacttac
     2161 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2221 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2281 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2341 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2401 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2461 gattgattta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2521 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     2581 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     2641 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     2701 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     2761 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
     2821 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     2881 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttaaaag gatctaggtg
     2941 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3001 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3061 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3121 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3181 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3241 tacctcgctc tgctaatcct gttaccagtc aggcatttga gaagcacacg gtcacactgc
     3301 ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct
     3361 gccctgaacc gacgaccggg tcgaatttgc tttcgaattt ctgccattca tccgcttatt
     3421 atcacttatt caggcgtagc aaccaggcgt ttaagggcac caataactgc cttaaaaaaa
     3481 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     3541 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     3601 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     3661 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     3721 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     3781 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     3841 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     3901 accgtctttc attgccatac ggaactccgg atgagcattc atcaggcggg caagaatgtg
     3961 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     4021 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     4081 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     4141 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     4201 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
     4261 aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa
     4321 gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg tcgggtgatg ctgccaactt
     4381 actgatttag tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct
     4441 gtccctcctg ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc
     4501 agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt
     4561 gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg
     4621 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcggcgagcg
     4681 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     4741 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     4801 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     4861 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     4921 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     4981 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     5041 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     5101 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     5161 aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga
     5221 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     5281 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa
     5341 atatttctag gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     5401 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     5461 cgtagaggat ctgctcatgt tt
//
