LOCUS       sequence-411032-        5460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISCath1 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5460)
  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 5460)
  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_ISCath1_reRNA
FEATURES             Location/Qualifiers
     source          1..5460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..39
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             55..66
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1492..1509)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1662..1748
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1840..1867
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1886..1977
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2435..2890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2522..2541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2732..2753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3446..4105)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4020..4039
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4106..4208)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4600..4617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4734..5279)
                     /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(4814..4833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5355..5374
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cgatctgaca gctagctcag tcctaggtat aatgctagcc ccgggaaaag atctaaagag
       61 gagaaaatga ttgtaaataa agcatataag tttcgtctct atccgaacaa agaacagcaa
      121 atattaatta ataaaacatt tggttgttct cgatttgtat ttaatcattt tttagcaaaa
      181 tgggataatg cttacaaaga aacaggaaaa ggattaactt ataattcttg ttctgctcag
      241 ttaacacaat taaaaaaaga gttaacttgg ctaaaagaag tagatagtat tgcgattcaa
      301 tcgtcattaa aaaatctcgc tgatgcttat tctcgtttct tcaagaaaca aaatgataga
      361 ccacgtttta agtctaaaaa aaataaagtg caatcctata caaccaaata tactaatggc
      421 aatattgcga ttaatgacaa taaaatcaaa ttgccgaaac ttggatggat tcgttttgtt
      481 aaaagtcgag aagttgaagg tcgcatttta aatgtaacca ttagacgaaa tccaagtggt
      541 aaatactttg tatctatttt agttgaaaca gaagttcaag aattatctaa aactaattct
      601 gctattggaa ttgatgtagg attaaaggat ttcgctattt tgtctaatgg aacgacttat
      661 gcaaatccta aattcttccg taaattagaa gaaaaattag caaaagcaca acgtattctt
      721 tctagacgta caaaaggtag ttctaattgg aataaacaac gaattaaagt tgcttgtatt
      781 catgaaaaaa tcacaaacgc aagaaacgat tacttacata aaatctctac cgaaattatc
      841 aaaaaccacg atattatcgg tatagaagat ttgcaagtaa gtaatatgtt gagaaatcat
      901 aatcttgcaa aagcgattag tgaagtatct tggtcacaat ttagaacgat gttggaatac
      961 aaagctaaat ggtatggaaa acaagtagtt accgtagcga aaaattttcc atccagccaa
     1021 ctttgttcat gctgtggcta tcaaaataaa gacgttaaaa atcctgggtt acgtgagtgg
     1081 gaatgtccaa agtgtcatac ccatcatgac cgagatatca acgcaagttt gaatcttaaa
     1141 aatgaagcca taaggcttct aaccgtagga actacgggga tagcctgaga gatctttgac
     1201 agctagctca gtcctaggta taatagttaa aaatcctggg ttacgtgagt gggaatgtcc
     1261 aaagtgtcat acccatcatg accgagatat caacgcaagt ttgaatctta aaaatgaagc
     1321 cataaggctt ctaaccgtag gaactacggg gatagcctaa tcaattagag ttcggtagaa
     1381 ctctgtactt aggaatccac tacggcttta gcctagtggt agttcaagct cggagatcat
     1441 cattgcgttt tttaagcttg gctgttttgg cggatgagag aagattttca gcctgataca
     1501 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
     1561 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     1621 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     1681 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     1741 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     1801 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
     1861 gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat
     1921 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     1981 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2041 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcagca
     2101 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2161 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2221 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2281 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2341 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2401 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc
     2461 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2521 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2581 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2641 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2701 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2761 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2821 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2881 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2941 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3001 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3061 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     3181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtcaggca
     3241 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     3301 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg
     3361 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag
     3421 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt
     3481 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     3541 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
     3601 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
     3661 gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
     3721 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
     3781 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
     3841 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaac tccggatgag
     3901 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     3961 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     4021 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     4081 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     4141 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     4201 gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga
     4261 caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa
     4321 gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct
     4381 ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt
     4441 aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg
     4501 gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt
     4561 gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg
     4621 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
     4681 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
     4741 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
     4801 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
     4861 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
     4921 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
     4981 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
     5041 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
     5101 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
     5161 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
     5221 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
     5281 aagatcatct tattaatcag ataaaatatt tctaggctca tgagcccgaa gtggcgagcc
     5341 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     5401 gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac agcttatcat
//
