LOCUS       sequence-410880-        5597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISCco1 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5597)
  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 5597)
  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_ISCco1_reRNA
FEATURES             Location/Qualifiers
     source          1..5597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..48
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             64..75
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1638..1655)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1808..1894
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1986..2013
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2032..2123
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2581..3036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2668..2687)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2878..2899
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3592..4251)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4166..4185
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4252..4354)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4746..4763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4880..5425)
                     /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(4960..4979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5501..5520
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcttatcatc gatctgacag ctagctcagt cctaggtata atgctagccc cgggaaaaga
       61 tctaaagagg agaaagaatt catgcaaata accaaaggct ttaagtatag gctatatcca
      121 aacaaagagc aaagcttaaa gctttctcat caatgtttca tctataatca agcctataat
      181 atttgcctaa acttacaaca agaacaatgg ctaaaaaata aagaattaga taaaaaagaa
      241 agaacttatt taagtgctac tcaaatcgat aatcaagtaa aactagcctt aaaagctaga
      301 aacttagagt ttaaaagtgt agtagcacaa caagctagaa taaatgctaa tagagcatta
      361 gaacaagcaa taaaagataa aaataaaggt tttccaaagt ttaaaaactc taaattggca
      421 aagcaaagtt tcaattggaa taatcaaggc tatcaaattc tagaatttaa cgataagttt
      481 aaaatattta gacttatgtc tatgcctata aaaatgcgtt atcatagagc tttgccaaat
      541 aattataaac taaatcaaat aactatttct aaaactcata ataaatactt tgtctcattt
      601 agtataactt atgatgaacc aagtgttgtt ttaatgtcta gcaataaaat agacattaac
      661 agagcagttg gcattgatat gaatatcaat gaaattgctt taagcaatgg tgctttgtta
      721 ctgacttgtt ctaagagtat aaataaagct aaatactcta ctactttcaa acgcttaatg
      781 cgaaaacaaa gtaaaagagt attgaaagct aaaaaggata agattaagtt aggttctaat
      841 tttagaaaaa ctcaaattag acttaataaa atctatgaaa gagctagtaa tataaaaaaa
      901 gatttagctc ataagataag taatcaaata gtaaaaaact ttgatttgat agtagtagaa
      961 aatctaaaag tgaaaaatat gactaaacga gcaaagctta aaaatgtaaa agctaaaagt
     1021 ggcttaaata agtctatatt agatacttct ttctatcaat ttttaagctt tgtggaatat
     1081 aaagctatgc ataatggcaa agtatttact aagatagaac cacaatatac aagcaaagct
     1141 tgttgcaaat gtggaactat aaaagagaat ttaactctaa aagatagagt ttttacttgc
     1201 gataattgtg gatatgtaga acatagagat ataaatgcaa gtaaaaatat cttggataga
     1261 ggcttgaagt cgtttgggct aggaactagc ctattagact ataaacaaga agccttgcgt
     1321 gttagcggat cctgagagat ctttgacagc tagctcagtc ctaggtataa tagatagagt
     1381 ttttacttgc gataattgtg gatatgtaga acatagagat ataaatgcaa gtaaaaatat
     1441 cttggataga ggcttgaagt cgtttgggct aggaactagc ctattagact ataaacaaga
     1501 agccttgcgt gttagctgac attggttggc tttagtaagg aagcacctac ttctataagt
     1561 aggtgtggtt cacgctcgga gatcatcatt gcgtttttta agcttggctg ttttggcgga
     1621 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     1681 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     1741 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     1801 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     1861 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     1921 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     1981 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     2041 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2101 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2161 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2221 atgctgaaga tcagttgggt gcagcaaact attaactggc gaactactta ctctagcttc
     2281 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2341 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2401 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2461 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2521 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2581 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2641 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2701 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2761 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     2821 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2881 gactcttgtt ccaaactgga acaacactca accctatctc gggctattct tttgatttat
     2941 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3001 acgcgaattt taacaaaata ttaacgttta caatttaaaa ggatctaggt gaagatcctt
     3061 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     3121 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3181 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3241 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     3301 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3361 ctgctaatcc tgttaccagt caggcatttg agaagcacac ggtcacactg cttccggtag
     3421 tcaataaacc ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac
     3481 cgacgaccgg gtcgaatttg ctttcgaatt tctgccattc atccgcttat tatcacttat
     3541 tcaggcgtag caaccaggcg tttaagggca ccaataactg ccttaaaaaa attacgcccc
     3601 gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc
     3661 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     3721 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     3781 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     3841 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     3901 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     3961 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     4021 cattgccata cggaactccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     4081 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     4141 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     4201 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     4261 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     4321 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc aaaagttggc
     4381 ccagggcttc ccggtatcaa cagggacacc aggatttatt tattctgcga agtgatcttc
     4441 cgtcacaggt atttattcgg cgcaaagtgc gtcgggtgat gctgccaact tactgattta
     4501 gtgtatgatg gtgtttttga ggtgctccag tggcttctgt ttctatcagc tgtccctcct
     4561 gttcagctac tgacggggtg gtgcgtaacg gcaaaagcac cgccggacat cagcgctagc
     4621 ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
     4681 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
     4741 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
     4801 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
     4861 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
     4921 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4981 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
     5041 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
     5101 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
     5161 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
     5221 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
     5281 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
     5341 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
     5401 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta
     5461 ggctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     5521 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     5581 tctgctcatg tttgaca
//
