LOCUS       sequence-410876-        5560 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISTva1 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5560)
  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 5560)
  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_ISTva1_reRNA
FEATURES             Location/Qualifiers
     source          1..5560
                     /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)"
     primer_bind     complement(1610..1627)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1780..1866
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1958..1985
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2004..2095
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2553..3008
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2640..2659)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2850..2871
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3564..4223)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4138..4157
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4224..4326)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4718..4735)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4852..5397)
                     /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(4932..4951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5473..5492
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgtttgacag cttatcatcg atctgacagc tagctcagtc ctaggtataa tgctagcccc
       61 gggaaaagat ctaaagagga gaaaatgcag agagcgttta aagttacact cattcctaat
      121 cacaatcaag aggtcttaat caataagagt attggttgtg caaggtttgt gtataaccac
      181 tttctggcat taaagcaaga gctatatcag tccgaacaga aaacattaaa ctataacgct
      241 tgtagtcaac gtttaacttt actcaagaaa gaaattgaat ggttgaagga agtagataag
      301 tttgccttac agaactcgct caagaattta gagacagcat acaaaaactt ttttgctgac
      361 ttaaagaagg tcaaaggcaa aaaaggagta ggctttccca agtttaaaaa gaagcatggt
      421 tgcaagcagt cttacaagac gaatttaaca aacggtaaca tccaggtaat agagaatcgt
      481 ttaaagcttc cgaagttagg gtgggtgaag tttcataagt cccaagaagt taccggaaag
      541 cttataaacg ttaccataac tcggacttct tctggtaaat atgttgctag tattctgtgt
      601 gatacagaga ttgagaaaca tccccaagtt agtcaaaata ttggcataga cttaggcata
      661 aagtcttatc tcgttactag taatggggaa gtcgtagata atcccaaata ttaccggact
      721 caaactcgta aattacgtaa agcacataaa aaattatccc gcagtgtaaa aggcagtact
      781 aatcgagtca aagcgaaaat caagctggct cgtagctacg aaagaattac caatctcaga
      841 gatgactttc tgcacaagct gtcaactcgt ctaatcaaag aaaacagtat tatctgtatt
      901 gaggatttgc gagttgccaa catggtaaaa aaccataaat tatcattgag tatttcagac
      961 gctagttggt ctaagttcgt tgccatgctt gaatataaag ctttttggca tgacagaatt
     1021 gtgcagaagg ttggtacgtt ttatccctcc tctcagactt gtaatcattg tggttttatc
     1081 aaccctttgg tcaaagattt aaagttacgt gaatgggctt gtcctggttg tagtggttac
     1141 aatttgagag acaataacgc agcgttgaac atattaggtg agggattgag attaatagcc
     1201 gccgtgggta tcccggaggc tctaaacgcc tgtggagaac tcgtcagtcc tggagcaatt
     1261 caggcagaga tcgttgaagc aggaatcacg cgacttcaag tcgtgtgaga gatctttgac
     1321 agctagctca gtcctaggta taataggctt gtcctggttg tagtggttac aatttgagag
     1381 acaataacgc agcgttgaac atattaggtg agggattgag attaatagcc gccgtgggta
     1441 tcccggaggc tctaaacgcc tgtggagaac tcgtcagtcc tggagcaatt caggcagaga
     1501 tcgttgaagc aggaatcacg cgacttcaag tcgtgtgagg ttcaagctcg gagatcatca
     1561 ttgcgttttt taagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     1621 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     1681 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     1741 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     1801 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     1861 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     1921 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     1981 ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc
     2041 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2101 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2161 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcagcaaa
     2221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2341 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2521 caagtttact catatatact ttagattgat ttacgcgccc tgtagcggcg cattaagcgc
     2581 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2641 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2701 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2761 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2821 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2881 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2941 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3001 tacaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3061 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3121 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3181 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3241 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3301 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtcaggcatt
     3361 tgagaagcac acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac
     3421 ataagcggct atttaacgac cctgccctga accgacgacc gggtcgaatt tgctttcgaa
     3481 tttctgccat tcatccgctt attatcactt attcaggcgt agcaaccagg cgtttaaggg
     3541 caccaataac tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     3601 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     3661 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     3721 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     3781 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     3841 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     3901 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     3961 acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaactc cggatgagca
     4021 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     4081 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     4141 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     4201 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     4261 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     4321 cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca
     4381 ccaggattta tttattctgc gaagtgatct tccgtcacag gtatttattc ggcgcaaagt
     4441 gcgtcgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt gaggtgctcc
     4501 agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg tggtgcgtaa
     4561 cggcaaaagc accgccggac atcagcgcta gcggagtgta tactggctta ctatgttggc
     4621 actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct gcaccggtgc
     4681 gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac tcgctacgct
     4741 cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag
     4801 atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag
     4861 gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc
     4921 gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt
     4981 tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac
     5041 gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc
     5101 ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa
     5161 gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg
     5221 aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca
     5281 agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc
     5341 aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa
     5401 gatcatctta ttaatcagat aaaatatttc taggctcatg agcccgaagt ggcgagcccg
     5461 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     5521 gatgccggcc acgatgcgtc cggcgtagag gatctgctca
//
