LOCUS       sequence-410923-        5763 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression of ISMmu1 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5763)
  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 5763)
  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_ISMmu1_reRNA
FEATURES             Location/Qualifiers
     source          1..5763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..53
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             69..80
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        1405..1439
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     primer_bind     complement(1809..1826)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1979..2065
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2157..2184
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2203..2294
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2752..3207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2839..2858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3049..3070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3763..4422)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4337..4356
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4423..4525)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4917..4934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5051..5596)
                     /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(5131..5150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5672..5691
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgacagctta tcatcgatct gacagctagc tcagtcctag gtataatgct agccccggga
       61 aaagacctaa agaggagaaa gaattcatgc tgtataatag aagtaggaag gaggcgaata
      121 aaatggaaca taagaagatg gcggattatg cctatcatta cggtctgaaa gtccgcatct
      181 accccagccg gcaacagaaa gagatgattc gacgcaatgc ggatgcggca cgctgcatct
      241 acaatgaaca tgtcgctgtg aacaaggaga tcgcacgatt cggcagacgg agcatctaca
      301 tcgacttcat tgagaagcgc atcgaggcac tgcgcgagcg aatccgcacg gtaacagcat
      361 tgaaagaccg ctatccatgg ctccggcatc ccgatatcga tgcgcagtcc attgccaatg
      421 cacggcagaa ctaccagcat gcttggaagc agttccgtgc cgtccatcat gctggtgtgc
      481 cgaacttcca caagaaaacg tccgtcgagt cgtatcagac gaatccgcat tacagcaagg
      541 gtacggtgat ttcgctgttt accagtaacg cccgcttcct cgacaagcat catctgatgc
      601 tgcccaagct caaacgcatc cgcttccaag gttcgcgtaa aatactggac cgactttttg
      661 ccatgaagga agtccgcatc ggaacgatta ccatcacgaa agacagcctg caccgctact
      721 ttgcgtcatt ccaactcggc tccgatgaac cattcgtgaa agccagcccg aaaacaaagg
      781 cagagattgg catcgacctg aacatcgaga acttctatgc cgactcgaat ggcgagattg
      841 tggccaatcc tcgttattac cgcaggcaga aacgacggct ggccaacgca cagcgcaagc
      901 tgagccgtcg gattgtccgt gccaagaaag agcatcgctc gctccgtgag gcaaagaacg
      961 tcgaaaaaca gcgcttgctc gtcgccaggc tcacggcaca agtcatgcag cgccgcaaag
     1021 atttcctgca ccgcacttcc gcgacactga tcaagaacca cgatctggtc gcagcagaag
     1081 aactacggag caagaatctg ctgaagaacc atgccctggc tatgagcatc caagacaatg
     1141 gctggcggat gttccttagt atgctgactt ataaggcagc actgtacggc aagaacttcg
     1201 tcaccgtcga cccgcggaat acgacgcaaa cctgccacgc ttgtggccat atcatgagcg
     1261 gtaaagagaa actgacgctc aaggaccgcg agtggatctg cccgaaatgt gggatacacc
     1321 accttcgcga tgtgaatgcg gccaagaaca tcctagcgcg agccaaagca aagctcgtat
     1381 ccatgcatgg atcctgagag atctttgaca gctagctcag tcctaggtat aatactagtg
     1441 cggtaaagag aaactgacgc tcaaggaccg cgagtggatc tgcccgaaat gtgggataca
     1501 ccaccttcgc gatgtgaatg cggccaagaa catcctagcg cgagccaaag caaagctcgt
     1561 atccatgcat tgatgcgaac ctaccttgtc cgtttggcaa cttgcggacg ctaatggctt
     1621 tggtaattac cggacctctg ctgtacctcg caagactgcg aatgcagtaa gcctggccta
     1681 tctaagcaaa ttgaggtatc gaaagattcc ctcaagctcg cgacttcagt cgcgggtagt
     1741 tgacgctcgg agatcatcat tgcgtttttt aagcttggct gttttggcgg atgagagaag
     1801 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
     1861 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
     1921 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
     1981 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     2041 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     2101 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     2161 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
     2221 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2281 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2341 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2401 atcagttggg tgcagcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2461 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2521 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2581 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2641 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2701 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct
     2761 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2821 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2881 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2941 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     3001 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     3061 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     3121 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     3181 ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat
     3241 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3301 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3361 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3421 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3481 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3541 ctgttaccag tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac
     3601 cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg
     3661 ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta
     3721 gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca
     3781 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     3841 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     3901 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     3961 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     4021 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     4081 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     4141 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     4201 acggaactcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     4261 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     4321 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     4381 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     4441 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     4501 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt
     4561 cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg
     4621 tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat
     4681 ggtgtttttg aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta
     4741 ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat
     4801 actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga
     4861 aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg
     4921 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     4981 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     5041 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     5101 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     5161 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     5221 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     5281 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     5341 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     5401 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     5461 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     5521 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     5581 accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct aggctcatga
     5641 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     5701 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat
     5761 gtt
//
