LOCUS       sequence-390269-        5507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YP_009665506 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5507)
  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 5507)
  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_009665506
FEATURES             Location/Qualifiers
     source          1..5507
                     /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             1846..1869
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1908..1925)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2078..2164
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2256..2283
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2302..2393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2394..2462,2463..3254)
                     /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(2612..2631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3425..4013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3914..3933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4167..4184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4199..4339
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4265..4284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4425..4447
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4603..5481)
                     /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 tccatggttc tgcgtgcgct ggacctgttt agcggtatcg gtggcattac ccacggcctg
      481 cgtgagatcg ttgaaccgat tgcgttcgtg gagaaaaacg acgaagcgcg tagctttctg
      541 aagaaaaagt atccggaaat tccggttttc gacgatgtgt gcagctttga tgcgaccaaa
      601 tggatcgaca agctggatat cattctggcg ggttggccgt gcaccggttt cagcaacgcg
      661 ggtaccaaaa ccggcttcag ccacgaggcg agcggtctgt ttaccgaagt tatccgtatt
      721 gtgaaagagt gccgtccgaa gtatgttttc ctggaaaaca gccacaccct gagcctgttt
      781 gagaacgtga acgtggttgt gaaggcgttc gacgagctgg gttacgattg ccgttggatt
      841 acctgccgtg cgacctgcgt tggtgcgctg caccagcgtc accgttggtt ttgcctggtt
      901 gtgcgtcgtg acattgaact ggatatcggt attccgtacg tggaacgttt cgactggatg
      961 accaacgagc cgccgcgtca gatccagaag agcaacaagg agaacatcct gcacattagc
     1021 ctggcgggta acgcggttgt gccggatcaa gttcgttatg cgatggacat cctgtgcagc
     1081 ttcgatacca acaaggaatt tagcctgagc agccgtgcga aaaacggttt cagcaagaac
     1141 ggcgtgatct ttacctacga cattgagcac ccggcgcgtg aaccgctgaa catcgttctg
     1201 accccgcgtg agaacgaaac cagctttgtg attagctgcg atccgaaaaa ggttctgacc
     1261 aaaagcgtga tccagaagtt ctggccgacc ccgctgcaca gctgcaaata tgcgaccaag
     1321 tgcccgcgta ccctgaccac ccgtgttagc aaaatgctga gcgcgtgcgt gggtttcagc
     1381 gagggtggcg acaaaaacgg ctacctgagc gcgaagtggg tgctgtggct gatgggctac
     1441 gattataagt acatgaccag cgaattccag catgatgaag cagtggataa caagtttaac
     1501 aaagaacagc agaacgcgtt ttatgagatc ttgcacttac cgaatctgaa cgaggaacag
     1561 cgcaatgcgt tcattcagtc ccttaaagac gatccatctc aaagcgcaaa tctgctggca
     1621 gaagccaaga aactgaacga tgctcaagct ccgaaagtcg acaacaaatt caacaaagag
     1681 caacagaatg cgttctacga aattctgcat ttaccgaacc tgaatgaaga acagcgtaac
     1741 gcctttatcc aatcgctgaa agatgatcct agtcagtcag ccaatctgtt ggcggaagcg
     1801 aagaaactca atgacgccca agctcccaaa gttgacgcga atagctggag ccatccgcag
     1861 tttgaaaagc tttagggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
     1921 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
     1981 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     2041 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     2101 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     2161 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg
     2221 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct
     2281 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc
     2341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     2641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     2941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3541 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4201 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     4261 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg
     4321 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4381 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4441 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt cgcgcgcgaa
     4501 ggcgaagcgg catgcataat gtgcctgtca aatggacgaa gcagggattc tgcaaaccct
     4561 atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca acttgacggc
     4621 tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca
     4681 ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt
     4741 ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc
     4801 gcgccagctt aagacgctaa tccctaactg ctggcggaaa aaatgtgaca gacgcgacgg
     4861 cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc
     4921 gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt
     4981 aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga
     5041 atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg
     5101 ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag
     5161 ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc
     5221 ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg
     5281 tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt
     5341 gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc
     5401 ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg
     5461 atcattttgc gcttcagcca tacttttcat actcccgcca ttcagag
//
