LOCUS       sequence-389730-        5495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YP_009701829 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5495)
  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 5495)
  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-YP_009701829
FEATURES             Location/Qualifiers
     source          1..5495
                     /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             1834..1857
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1896..1913)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2066..2152
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2244..2271
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2290..2381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2382..2450,2451..3242)
                     /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(2600..2619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3413..4001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3902..3921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4155..4172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4187..4327
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4253..4272)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4413..4435
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4591..5469)
                     /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 tccatgctac acgctataga tttattttca ggaatcggcg gtattaccca tggtttgcgt
      481 ggtattgtgg aacctattgc gtatgtagaa aaaaatgatg acgccagagg cttcctcgcg
      541 cgtaaacatc cgaatgttcc ggtgtttgac gacgtttgca cctttgacgc caccccgtat
      601 ttgggcaaag ttgatattat cactggcggc tggccgtgca cgggtttcag caccgcaggc
      661 aagggaaccg gtttcgagca cgaggcctcc ggtttattca ccgaagtggt gcgcatcacg
      721 aaagagtgcc agccgaaata cttgttcttg gagaactctc acaccctggc ggcgtacgag
      781 aacatcaacg ttatcgtgaa ggcgtttgac gacatgggct acgattgccg ttggaccagc
      841 tgtcgtgcta cgtgcgtggg tgctccgcat caacgttacc gctggttttg tctggttgtg
      901 aaaaaaggcg ctggcatcga ctttgagatc ccggtcatcg ataaattcga ctgggaaaac
      961 aacgagccac cgcgccagat tgaaaaaaac aacaagacca ataagctgcg tatcggtttt
     1021 atgggcaatg cggttgttcc ggatcaagtt cgctacgcaa tgacccttct gtcgacgctt
     1081 gagaacaagg ttctgggtcc gagcaacacc gatggttaca gcattgacgg tcgtatctat
     1141 accttcgtgg taaaacaccc aacccgtaaa ccgctgaata ttgtgctgac gccgcgtgaa
     1201 aacgaagcga gctttgcgaa aatctgcgat ccgaagaagg tcctgaccaa gccagtcacc
     1261 aagaagtatt gggcaactcc ggtgtataat tgtatgaaca gcgcgaagtg cccgcgtacc
     1321 ctgaccaagc gcgtcagcaa catgctgtcg gcatgtgttg ggttctctga aggtggcaac
     1381 aaaaactggt atatgaacgc cgattggatt atgtggctga tgggttacga gccgggttac
     1441 ctgagccacg aattccagca tgatgaagca gtggataaca agtttaacaa agaacagcag
     1501 aacgcgtttt atgagatctt gcacttaccg aatctgaacg aggaacagcg caatgcgttc
     1561 attcagtccc ttaaagacga tccatctcaa agcgcaaatc tgctggcaga agccaagaaa
     1621 ctgaacgatg ctcaagctcc gaaagtcgac aacaaattca acaaagagca acagaatgcg
     1681 ttctacgaaa ttctgcattt accgaacctg aatgaagaac agcgtaacgc ctttatccaa
     1741 tcgctgaaag atgatcctag tcagtcagcc aatctgttgg cggaagcgaa gaaactcaat
     1801 gacgcccaag ctcccaaagt tgacgcgaat agctggagcc atccgcagtt tgaaaagctt
     1861 tagggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     1921 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     1981 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     2041 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2101 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     2161 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     2221 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     2281 acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2341 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2401 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2461 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2521 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2581 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     2641 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2701 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2761 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2821 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2881 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2941 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3001 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3061 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3121 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3181 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3241 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3301 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3361 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3541 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3781 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4081 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4141 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4201 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
     4261 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4321 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4381 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4441 accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca
     4501 tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt
     4561 caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac
     4621 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
     4681 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
     4741 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
     4801 gacgctaatc cctaactgct ggcggaaaaa atgtgacaga cgcgacggcg acaagcaaac
     4861 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
     4921 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
     4981 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
     5041 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
     5101 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
     5161 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
     5221 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa
     5281 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
     5341 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
     5401 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
     5461 ttcagccata cttttcatac tcccgccatt cagag
//
