LOCUS       sequence-410906-        5566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISTfu1 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5566)
  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 5566)
  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_ISTfu1_reRNA
FEATURES             Location/Qualifiers
     source          1..5566
                     /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)"
     promoter        1284..1318
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1602..1619)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1772..1858
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1950..1977
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1996..2087
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2545..3000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2632..2651)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2842..2863
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3556..4215)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4130..4149
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4216..4318)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4710..4727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4844..5389)
                     /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(4924..4943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5465..5484
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcatcgatct gacagctagc tcagtcctag gtataatgct agccccggga aaagatctaa
       61 agaggagaaa gaattcatgc tgaccggacg ccgctacctg ctggctttca cccccgagca
      121 ggaggagttc gccgagaaga tcggtgacgc ctgccgggtt gtgtggaaca ccgcgctgga
      181 gcagcgccga cagtaccggc ggcgcggcgc gttcatcggt tacgtagagc aggcgcggca
      241 gatggccgag gccaagaagg acttcccgtg gctggccgag gccccgtcgc acacgcttca
      301 gcaaacgctc cgcgaccttg agcgggcatg caagacgcac ggcacgttca aggtccgctg
      361 gcgctccaag cggaaaaacg cgccgacgtt ccgcttcccc gatcccaagc agatcaccgt
      421 ggagcgaatc tcccgtcgct ggggccgggt caagctgccc aagctgggct ggacccgctt
      481 ccggtggaca cgcccgctcg gcgggcaact tcgcaatgcc accgtgctca aggacggtgg
      541 gcgctggtac atctccttct gcgtcgaaga cgggctgatg gagtccgcac cgaatggcaa
      601 acccccggtg ggggtggatc gtggtgtcac tgtcgcagtc gccacctcgt ccgggtggat
      661 gcgtgaccgc gagttcgtca ccccagggga agcggtacgg ctcaagcgtc ttcagcagca
      721 gctcgcccgg caacggaagg gctccaaccg gcgcgcggcc acccgggcga agctgaccaa
      781 gctgaacgcc cgcatccgcg cccggcgtac cgacttcctg gcgtggaccg ccaaccgcct
      841 cacctgtgac cacggactgg tcgttgtgga ggacctcaac atccgaaaca tgaccgcctc
      901 tgccaaaggc acgcttgccg agccggggag gaacgtcgcc gccaaggctg ggctgaaccg
      961 ggcgatcctg gccaagggat ggggcggact actgaccgcc ttggagcaca aggctcgtta
     1021 ccacggctcc cgtattctgc gggtgccacc tgcgttcaca tcgcagacct gccatgcctg
     1081 cgggcactgc gcgccggaca accgtgaaag ccaagcggag ttccggtgcc gcgcctgcgg
     1141 ccaccaggcc aacgccgacg tgaacgccgc gaagaacatc ctcgccgccg ggctggcggt
     1201 gacagggcgt ggagacctcg ccgttgggcg gtctgtgaag cgccaaccac ccaaggtaga
     1261 ggcagcggga tcctgagaga tctttgacag ctagctcagt cctaggtata atactagtgt
     1321 gaaagccaag cggagttccg gtgccgcgcc tgcggccacc aggccaacgc cgacgtgaac
     1381 gccgcgaaga acatcctcgc cgccgggctg gcggtgacag ggcgtggaga cctcgccgtt
     1441 gggcggtctg tgaagcgcca accacccaag gtagaggcag cgtgaacgcg cctcagccac
     1501 gggaatcccc cgccttcagg cgtggggagg aagtcaagct cggagatcat cattgcgttt
     1561 tttaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     1621 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     1681 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     1741 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     1801 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     1861 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     1921 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     1981 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2041 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2101 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2161 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcagca aactattaac
     2221 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2281 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2341 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2401 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2461 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2521 ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2581 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2641 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2701 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2761 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2821 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2881 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2941 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     3001 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3061 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3121 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3181 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3241 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3301 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtcaggca tttgagaagc
     3361 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
     3421 ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc
     3481 attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag ggcaccaata
     3541 actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
     3601 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
     3661 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     3721 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     3781 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     3841 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     3901 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     3961 ccatatcacc agctcaccgt ctttcattgc catacggaac tccggatgag cattcatcag
     4021 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     4081 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     4141 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     4201 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     4261 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
     4321 gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
     4381 tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg
     4441 tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt
     4501 ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa
     4561 gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga
     4621 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
     4681 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
     4741 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
     4801 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
     4861 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
     4921 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
     4981 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
     5041 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
     5101 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
     5161 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
     5221 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt
     5281 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
     5341 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     5401 tattaatcag ataaaatatt tctaggctca tgagcccgaa gtggcgagcc cgatcttccc
     5461 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     5521 ccacgatgcg tccggcgtag aggatctgct catgtttgac agctta
//
