LOCUS       sequence-417996-        5584 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression of ISAba30 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5584)
  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 5584)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_ISAba30_reRNA
FEATURES             Location/Qualifiers
     source          1..5584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(948..1403)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1085..1106)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1297..1316
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        complement(1861..1952)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1971..1998)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2090..2176)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2329..2346
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     promoter        complement(2615..2649)
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     RBS             3896..3907
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3923..3957)
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     primer_bind     complement(4048..4067)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     rep_origin      4143..4688
                     /direction=RIGHT
                     /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     4589..4608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4805..4822
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5214..5316
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(5317..5584,1..392)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5383..5402)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
       61 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
      121 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct
      181 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
      241 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
      301 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
      361 agtactgcga tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt
      421 aaacgcctgg ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga
      481 aagcaaattc gacccggtcg tcggttcagg gcagggtcgt taaatagccg cttatgtcta
      541 ttgctggttt accggtttat tgactaccgg aagcagtgtg accgtgtgct tctcaaatgc
      601 ctgactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      661 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
      721 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      781 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      841 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      901 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttgtaaacgt
      961 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     1021 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag ggttgagtgt
     1081 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     1141 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt
     1201 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     1261 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     1321 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     1381 taatgcgccg ctacagggcg cgtaaatcaa tctaaagtat atatgagtaa acttggtctg
     1441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1741 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1801 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1861 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1921 catatttgaa tgtatttaga aaaataaaca aaagagtttg tagaaacgca aaaaggccat
     1981 ccgtcaggat ggccttctgc ttaatttgat gcctggcagt ttatggcggg cgtcctgccc
     2041 gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga tttgtcctac
     2101 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
     2161 cctttcgttt tatttgatgc ctggcagttc cctactctcg catggggaga ccccacacta
     2221 ccatcggcgc tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg gaccaccgcg
     2281 ctactgccgc caggcaaatt ctgttttatc agaccgcttc tgcgttctga tttaatctgt
     2341 atcaggctga aaatcttctc tcatccgcca aaacagccaa gcttaaaaaa cgcaatgatg
     2401 atctccgagc ttgacatcct ccccggcctg aaggccggag attcctacgg cgctcagacg
     2461 cggcaatgag cctggtctga gtcgcttcgg tgggttcctg ctgctggcga ccctactgca
     2521 tcgctcactt cacaggctaa ccgggcgtgc cccgccctta gtacattgat cgcgccgacc
     2581 acatcggcat taccctcgaa cccgcactct atgcactagt attataccta ggactgagct
     2641 agctgtcaaa gatctctcag gatccgacgc ggcaatgagc ctggtctgag tcgcttcggt
     2701 gggttcctgc tgctggcgac cctactgcat cgctcacttc acaggctaac cgggcgtgcc
     2761 ccgcccttag tacattgatc gcgccgacca catcggcatt accctcgaac ccgcactcta
     2821 tgcactcgaa ccgctcttgt gtctggcgat tatccgccga cacatgagca caacacgggc
     2881 aggttcggct ggtgttctta ggcggcacgg caacgagcca cccaccattc cacgccagtt
     2941 tataatccag ttggcgacgg aactcgaacc agccctgatc gaggatggat ttgttcaggc
     3001 cggacttggc ccgaacgttt cttcccggct gttcgggagt acccgccgcc gacttggaca
     3061 tgttccgtac ctgcaagtcc tcgatacaca ccatcgcgtg gttttggctg atcgcggtcg
     3121 aggtcttgtg caggaagtcg cggcgggcgt tgccgatgcg ggaatggatg cgctggatac
     3181 gggctttcgc cttctgccag ttgttgctga atttcacctt gcgggagagc gcttgctgcg
     3241 cgcagcgcaa gcgcttttcg tgtcgcttga agctgtttag aggagcgtag aacttgccgt
     3301 ccgagatcgt agcgaaccgg gcaatgccca tgtcgatacc gaccgcgcca ccctgcggga
     3361 tgggctgctc gacctctcgt tcggtttgaa tggacacgaa ccacttgccg cccgagaggc
     3421 tgacagtgac gttcttcacc gcccccagca catcgcggct cttgcggtag cgaatccagc
     3481 caagcttggg cagaaagatg cggctgttgt gctgctccag cttgatttgt ttcggatcgg
     3541 gatagcgaaa gctactgtgg caccccttct ttttgaagcg cgggaagtcg gcgcgcttgg
     3601 caaagaagtt ggaatacgcc cgctccaagt ccttgagcgt ctgttgcagc gggtgaaccg
     3661 gcgcatcagc caaccatgcc atttccgagc tattgcgcca ctcggtgagt agcttgcaca
     3721 gcccggcata gccgagcttc ttctcaccgc gctcgtagcg ctccttctgc aacgccagtg
     3781 ccttgttgta gacgaaccgg cacgagccag cgaagcggcg catctgccgt tcctgctggc
     3841 cgtttggtac catttcgtac ttgaaggctt gaagtcgttt catgaattcg aattctttct
     3901 cctctttaga tcttttcccg gggctagcat tatacctagg actgagctag ctgtcagatc
     3961 gatgataagc tgtcaaacat gagcagatcc tctacgccgg acgcatcgtg gccggcatca
     4021 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     4081 gggctcgcca cttcgggctc atgagcctag aaatatttta tctgattaat aagatgatct
     4141 tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg
     4201 cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt
     4261 ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc
     4321 aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct
     4381 ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg
     4441 gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg
     4501 tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa
     4561 caggagagcg cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg
     4621 ggtttcgcca ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc
     4681 tatggaaaaa cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc
     4741 ttccaggaaa tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga
     4801 gcgtagcgag tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg
     4861 caccggtgca gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg
     4921 ccaacatagt aagccagtat acactccgct agcgctgatg tccggcggtg cttttgccgt
     4981 tacgcaccac cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg
     5041 gagcacctca aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca
     5101 ctttgcgccg aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt
     5161 gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg
     5221 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     5281 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
     5341 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     5401 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     5461 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     5521 gctcatccgg agttccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     5581 cacc
//
