LOCUS       sequence-389728-        5522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YP_009702000 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5522)
  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 5522)
  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_009702000
FEATURES             Location/Qualifiers
     source          1..5522
                     /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             1861..1884
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1923..1940)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2093..2179
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2271..2298
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2317..2408
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2409..2477,2478..3269)
                     /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(2627..2646)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3440..4028
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3929..3948
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4182..4199
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4214..4354
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4280..4299)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4440..4462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4618..5496)
                     /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 tccatgctgc gtgcgctgga cctgttcagc ggcatcggtg gcattaccta cggcctgcgt
      481 ggtatcgtta ccccggtggc gtatgttgag aagaacaaag acgcgcgtga atttctgcag
      541 aagaaacacc cggatgttcc ggtgttcgac gatgtttgcg cgtttgacgc gaccgagtgg
      601 aagggcaaag tggatatcat taccggtggc tggccgtgca ccggtttcag catcgcgggt
      661 aaaggcgcgg gtttcgagca cgaagcgagc ggtctgttta gcgaggtggt tcgtatcacc
      721 aacgagtgcg aaccgcgtta cctgttcctg gaaaacagcc acattctgag caagaaaacc
      781 aacgttagcg tggttgtgaa cgcgctggac aagctgggct acgattgcaa atggctgacc
      841 tgccgtgcga ccaacgtggg tgcgccgcac caacgtcacc gttggttctg cctggtgatc
      901 aagcgtggcg cgagcgttat taccgacctg ccgctggttg acacctttaa ctggagcagc
      961 ggcgagccgg acaagcagat caaaaactat accccgatta accgtaccgt tagcagctgg
     1021 ctgggtaacg cggttgtgcc ggaccaagtg cgttacgcgt tcaccaccct gaacaccatg
     1081 agcaccgaaa tcgcgaccta tcagcacgat attaacaagt tccaaaactg cggttttagc
     1141 ctgaaaagcg agatctactg gagccgtatc aagattcagg aacgtccgcc gctgtatatt
     1201 accctgaccc aagagaaccc gccgaacaag cacaaagcga ccaccccgca catcaaaacc
     1261 attaccaaga aattctggag caccccggtt tttagccaca acccgagcgg cgtgaacttc
     1321 ctgacctacc gtagcagcaa gctgctgggc acccaggtta aattcgcgga aggtggcttt
     1381 gcgggcatgt atgtgaaccc ggattgggtt cgttggctga tgggttaccc ggaggactat
     1441 tttaccctgg cgtacaagga tcacgaaacc caaagcgaat tccagcatga tgaagcagtg
     1501 gataacaagt ttaacaaaga acagcagaac gcgttttatg agatcttgca cttaccgaat
     1561 ctgaacgagg aacagcgcaa tgcgttcatt cagtccctta aagacgatcc atctcaaagc
     1621 gcaaatctgc tggcagaagc caagaaactg aacgatgctc aagctccgaa agtcgacaac
     1681 aaattcaaca aagagcaaca gaatgcgttc tacgaaattc tgcatttacc gaacctgaat
     1741 gaagaacagc gtaacgcctt tatccaatcg ctgaaagatg atcctagtca gtcagccaat
     1801 ctgttggcgg aagcgaagaa actcaatgac gcccaagctc ccaaagttga cgcgaatagc
     1861 tggagccatc cgcagtttga aaagctttag ggctgttttg gcggatgaga gaagattttc
     1921 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc
     1981 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     2041 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     2101 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2161 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     2221 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     2281 cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt
     2341 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2401 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2461 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2521 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2581 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2641 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2701 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2761 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2821 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2881 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2941 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3001 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3061 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3121 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3181 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3241 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3301 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3361 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3421 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3481 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3541 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3601 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3661 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3721 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3781 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3841 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3901 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3961 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4021 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4081 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4141 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4201 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4261 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     4321 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     4381 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4441 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat
     4501 caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg acgaagcagg
     4561 gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta
     4621 tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg
     4681 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
     4741 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
     4801 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaaaatg
     4861 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
     4921 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
     4981 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
     5041 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
     5101 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
     5161 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
     5221 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
     5281 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
     5341 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
     5401 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
     5461 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag
     5521 ag
//
