LOCUS       sequence-390270-        5513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express M.CviPII (NDel15) in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5513)
  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 5513)
  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.CviPII (NDel15)
FEATURES             Location/Qualifiers
     source          1..5513
                     /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             1852..1875
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1914..1931)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2084..2170
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2262..2289
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2308..2399
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2400..2468,2469..3260)
                     /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(2618..2637)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3431..4019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3920..3939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4173..4190
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4205..4345
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4271..4290)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4431..4453
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4609..5487)
                     /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 tccatgaaag ccttggaact atttgcgggg ataggtggca ttacccatgg cttacgtggc
      481 tatgtagaac cgattgcatt ctgcgaatat gagaaagatg ccagctcgtt tctgtcacaa
      541 cgtggcttac ccgttcatgg ggacattacg aagtttgacg ccagtgtgta caaaaacaag
      601 attgacattg tgacagcagg atggccttgt actgggtttt cgaccgcggg taaaggcact
      661 ggttttgagc atgaagctag cggtttatgg acagaagtgg tccgagtagt gaaagagagc
      721 gaaccgaaat acgtcttttt ggaaaatagt cacgttttgg cgcaaaccaa aaacctcaag
      781 gtcatcatcc atgatctgga tatccttggc tatgatacgc gttggtggac gtgcagatcg
      841 aatgacgtta atgtcggtgc acatcacaat cggtatcgct ggtttatgct cgcagagaaa
      901 aaaggctctg tgaccaaatt cgtgaaaatt caggtgaaaa agttcaattg gtctggtgac
      961 ttcaaagaga aacagataag cgagaacagt cacgaaaaca agcagctcat taagttcatg
     1021 ggaaactccg tagttccgga tcaagttcgc tatgcgtttg aatcgatgag cgacatgatt
     1081 ctggaaggca gtctggtgaa tgataaggat gagatcgtga aggtaggcta ttccaaagat
     1141 ggcatcatgt acaaaattcc gattgaacac aaaatcattc cgaaacttaa catcgttctg
     1201 accccacgtg atccgccaga aggccataaa gcccgtgaag aagcgatcat caaaagcccc
     1261 attctgatga cgtactggaa tactccggcc ttttgctatc acaaatcagc tcgcggcgct
     1321 aaaattctga ccaaacgtca gaaaaacaac ctgcataccc agatcaaatt ctgtcctggt
     1381 ggttctgacg atggttatct gtcaggtcgc ttttgtgcgt ggctgatggg atacgatgat
     1441 gaatacctgg gggatcttat gcactatgaa ttccagcatg atgaagcagt ggataacaag
     1501 tttaacaaag aacagcagaa cgcgttttat gagatcttgc acttaccgaa tctgaacgag
     1561 gaacagcgca atgcgttcat tcagtccctt aaagacgatc catctcaaag cgcaaatctg
     1621 ctggcagaag ccaagaaact gaacgatgct caagctccga aagtcgacaa caaattcaac
     1681 aaagagcaac agaatgcgtt ctacgaaatt ctgcatttac cgaacctgaa tgaagaacag
     1741 cgtaacgcct ttatccaatc gctgaaagat gatcctagtc agtcagccaa tctgttggcg
     1801 gaagcgaaga aactcaatga cgcccaagct cccaaagttg acgcgaatag ctggagccat
     1861 ccgcagtttg aaaagcttta gggctgtttt ggcggatgag agaagatttt cagcctgata
     1921 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     1981 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     2041 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     2101 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     2161 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     2221 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     2281 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     2341 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2401 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2461 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2521 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2581 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2641 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2701 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2761 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2821 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2881 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2941 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3001 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3061 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3121 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3181 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3241 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3301 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3361 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3421 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3481 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3541 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3601 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3661 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3721 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3781 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3841 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3901 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3961 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4021 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4081 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4141 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4201 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4261 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     4321 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     4381 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4441 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
     4501 cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
     4561 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
     4621 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     4681 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     4741 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     4801 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaaaat gtgacagacg
     4861 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     4921 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     4981 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     5041 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     5101 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     5161 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     5221 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     5281 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     5341 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     5401 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     5461 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gag
//
