LOCUS       sequence-390281-        5555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express M.CviPI in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5555)
  AUTHORS   Miura F, Miura M, Shibata Y, Furuta Y, Miyamura K, Ino Y, Bayoumi 
            AMA, Oba U, Ito T
  TITLE     Identification, expression, and purification of DNA cytosine 
            5-methyltransferases with short recognition sequences.
  JOURNAL   BMC Biotechnol. 2022 Nov 4;22(1):33. doi: 
            10.1186/s12896-022-00765-3.
  PUBMED    36333700
REFERENCE   2  (bases 1 to 5555)
  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: "BMC 
            Biotechnol."; date: "2022-11-4"; volume: "22(1):33. doi"; pages: " 
            10.1186/s12896-022-00765-3"
COMMENT     Sequence Label: pBZS-M.CviPI
FEATURES             Location/Qualifiers
     source          1..5555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1894..1917
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1956..1973)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2126..2212
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2304..2331
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2350..2441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2442..2510,2511..3302)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2660..2679)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3473..4061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3962..3981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4215..4232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4247..4387
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4313..4332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4473..4495
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4651..5529)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHFFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagccg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa gctcgaggga
      421 tccatgaccc tgaaggccct ggagctgttc gccggcatcg ccggcatcac ccacggcctg
      481 agaggcttcg tggagcccgt ggccttcgtg gagatcaaca aggacgccca ggagttcctg
      541 agcaccaagt tccccgacaa gcccgtgttc gacgacgtga ccaagttcag caagagagac
      601 ttcgacgagc ccatcgacat gatcaccggc ggcttcccct gcaccggctt cagcatcgcc
      661 ggcaagagaa acggcttcga gcacgccgag agcggcctgt tcggcgaggt ggtgagaatc
      721 accaaggagt acatgcccaa gatggtgttc ctggagaaca gcggcatgct gagccacaag
      781 tacaacctgg acatcgtgat cagaagcatg gacagcctgg gctacgactg cagatgggtg
      841 accctgagag ccaccgtggt gggcgccctg cacaccagac acagatggtt ctgcctgtgc
      901 accagaaagg accacatcag agagaccctg atctgcgaca gagaggtgac caagttcgac
      961 tgggagaacg acagaccccc catccaggtg gacagcagaa gctacgagaa cagcagactg
     1021 gtgagattcg ccggctacag cgtggtgccc gaccagatca gatacgcctt caccggcctg
     1081 tacaccggca acttcagccc cagcttcagc aagaccctgg tgcccggcag cctggagggc
     1141 agcatctgct tcaacgagga caagatcacc aacggctact acaaggacgg cgtgtactac
     1201 gagttcgtga gaaccgagac ccacagagag cccgtgaaca tcctgctgac ccccagagag
     1261 atccccaaca agcacaacgg caagaagctg ctgaccctgc ccgtgaccaa gagatactgg
     1321 tgcaccccct gcgccagcta cggcaagggc accgccggcg gcagagtgct gaccgacaga
     1381 agcagccaca gcctgcccac ccaggtgaag ttcagccccg agggcgagga cggcaagcac
     1441 ctgagcggca agttctgcgc ctggctgatg ggctacgaca aggagtacct gggcaacctg
     1501 ctggagtacg aattccagca tgatgaagca gtggataaca agtttaacaa agaacagcag
     1561 aacgcgtttt atgagatctt gcacttaccg aatctgaacg aggaacagcg caatgcgttc
     1621 attcagtccc ttaaagacga tccatctcaa agcgcaaatc tgctggcaga agccaagaaa
     1681 ctgaacgatg ctcaagctcc gaaagtcgac aacaaattca acaaagagca acagaatgcg
     1741 ttctacgaaa ttctgcattt accgaacctg aatgaagaac agcgtaacgc ctttatccaa
     1801 tcgctgaaag atgatcctag tcagtcagcc aatctgttgg cggaagcgaa gaaactcaat
     1861 gacgcccaag ctcccaaagt tgacgcgaat agctggagcc atccgcagtt tgaaaagctt
     1921 tagggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     1981 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     2041 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     2101 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2161 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     2221 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     2281 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     2341 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2401 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2461 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2521 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2581 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2641 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     2701 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2761 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2821 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2881 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2941 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3001 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3061 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3121 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3181 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3241 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3301 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3361 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3421 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3481 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3541 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3601 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3661 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3721 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3781 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3841 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3901 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3961 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4021 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4081 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4141 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4201 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4261 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     4321 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4381 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4441 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4501 accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca
     4561 tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt
     4621 caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac
     4681 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
     4741 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
     4801 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
     4861 gacgctaatc cctaactgct ggcggaaaaa atgtgacaga cgcgacggcg acaagcaaac
     4921 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
     4981 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
     5041 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
     5101 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
     5161 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
     5221 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
     5281 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa
     5341 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
     5401 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
     5461 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
     5521 ttcagccata cttttcatac tcccgccatt cagag
//
