LOCUS       sequence-390273-        5576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YP_001497893 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5576)
  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 5576)
  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_001497893
FEATURES             Location/Qualifiers
     source          1..5576
                     /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             1915..1938
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1977..1994)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2147..2233
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2325..2352
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2371..2462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2463..2531,2532..3323)
                     /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(2681..2700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3494..4082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3983..4002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4236..4253
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4268..4408
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4334..4353)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4494..4516
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4672..5550)
                     /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 tccatggtac taagggcttt agatttgttt tcaggtatcg gcggtattac ccacggactg
      481 cgtgagattg tggagccgat tgcattcgtg gagaaaaacg atgaggcgcg tagctttctg
      541 aagaagaaac atccggaaat cccggttttt gacgatgttt gttctttcga cgcgactaag
      601 tggacccatg ttgatatcat tctggctggc tggccgtgta ccggtttctc taatgctggc
      661 accaaaaccg gtttctccca cgaagcgagc ggtttgttca ccgaagttgt gcgcataacc
      721 gaagagtgcc gtccgaagta cgtttttctg gaaaacagcc acaccctgag cttgttcgag
      781 aacatttccg tgatcgtgaa tgcctttgat gaactgggtt acgactgtcg ttggattacg
      841 tgcagagcga cgtgcgtggg cgcactgcac cagcgtcatc gttggttttg cctggtcgtc
      901 cgccgtgata ttgagctgga catcggtatc ccgtatgtgg agcgctttga ctggatgacc
      961 aacgaaccgc ctcgccagat tcaaaaaagc aataaggaaa acatcctgca tattagctta
     1021 gccggcaacg cggttgttcc ggatcaggtt cgctatgcta tggatatcct ttgctccttt
     1081 gacaccaaca aagagttctc cttgtcgagc cgtgcgaaaa atggcttcag taagaacggc
     1141 gttattttta cctatgacat tgagcacccg gcgcgtgagc cgctgaatat cgtgttgacg
     1201 ccacgtgaga acgagaccag cttcgtcatc tcttgtgatc cgaaaaaggt gctcacgaag
     1261 tcggtcatcc aaaagttttg gccaacgccg ctgcacagct gcaaatacgc taccaagtgc
     1321 ccgagaactc ttaccacccg tgtgagcaag atgctgtctg cgtgcgttgg tttcagcgaa
     1381 ggtggcgata aaaacggcta cctgtccgcg aaatgggttc tgtggctgat gggttacgac
     1441 gactcataca tgagcaccgc agaaagcaag tatgaaatga atcgcgcctt ctttctggac
     1501 atcatcgtag gtgaacgtgc gagcgtgttc gaattccagc atgatgaagc agtggataac
     1561 aagtttaaca aagaacagca gaacgcgttt tatgagatct tgcacttacc gaatctgaac
     1621 gaggaacagc gcaatgcgtt cattcagtcc cttaaagacg atccatctca aagcgcaaat
     1681 ctgctggcag aagccaagaa actgaacgat gctcaagctc cgaaagtcga caacaaattc
     1741 aacaaagagc aacagaatgc gttctacgaa attctgcatt taccgaacct gaatgaagaa
     1801 cagcgtaacg cctttatcca atcgctgaaa gatgatccta gtcagtcagc caatctgttg
     1861 gcggaagcga agaaactcaa tgacgcccaa gctcccaaag ttgacgcgaa tagctggagc
     1921 catccgcagt ttgaaaagct ttagggctgt tttggcggat gagagaagat tttcagcctg
     1981 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     2041 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     2101 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     2161 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     2221 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     2281 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     2341 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
     2401 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2461 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2521 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2581 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2641 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2701 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2761 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2821 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2881 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2941 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3001 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3061 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3121 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3181 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3241 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3301 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3361 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3421 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3481 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3541 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3601 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3661 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3721 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3781 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3841 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3901 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3961 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     4021 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4081 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4141 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4201 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4261 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     4321 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     4381 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     4441 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4501 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc
     4561 gcgcgcgaag gcgaagcggc atgcataatg tgcctgtcaa atggacgaag cagggattct
     4621 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
     4681 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     4741 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     4801 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     4861 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa aatgtgacag
     4921 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     4981 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     5041 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     5101 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     5161 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     5221 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     5281 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     5341 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     5401 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     5461 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     5521 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagag
//
