LOCUS       sequence-390272-        5516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YP_001426521 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5516)
  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 5516)
  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_001426521
FEATURES             Location/Qualifiers
     source          1..5516
                     /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             1855..1878
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1917..1934)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2087..2173
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2265..2292
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2311..2402
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2403..2471,2472..3263)
                     /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(2621..2640)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3434..4022
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3923..3942
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4176..4193
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4208..4348
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4274..4293)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4434..4456
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4612..5490)
                     /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 tccatgaagg cgctggaact gttcgcgggt gtgggtggca ttacccacgg cctgcgtggt
      481 tacgttaagc cggtggcgtt cgttgagtat gaaaaagatg cggcggagtt tctggcggcg
      541 cgtggcaagc cggtgcacgg tgacgttaaa gagttcgatg cgaccgaata ccgtggcaac
      601 atcgatattg ttaccgcggg ctggccgtgc accggtttta gcaccggtgg caaaggtggc
      661 ggtttcgacc acgaggcgag cggtctgttt gtggaagtgg ttcgtgtggt tcgtgagtgc
      721 gatccgcgtt acgttttcct ggaaaacagc cacgtgctga gccaggttcc gaacctgagc
      781 gtggttgtga gcgagctgca cgcgctgggc tatgactgcc gttggtacag ctgctatagc
      841 aacgatgtgt gcatcggtgc gccgcaccaa cgtcaccgtt ggttctgcct ggcgtttaag
      901 cgtgacctgg gcatcagcat tccgaagacc tacgcgccga aatttgattg gagcagcgaa
      961 cacccgtgca cccaggaagt ggaaaacacc cgtgttaaca agcgtctgct gaaactgatg
     1021 ggtaacagcg ttgtgccgag ccaagtgcgt cacagcttcg agatgatgct gggcatggaa
     1081 aaaaccggtg ttccggttgg cagcggtgat gttcatgcgc gttgcggcta tgcggaaaac
     1141 ggtattatgt ttcgtgtgga gatcaaccag cgtaagattg cgccggttaa catcctgctg
     1201 gagccgaaag aagtgccgaa gaaacaccgt gttccggata gccgtaacat cctgaccaac
     1261 gcggtgcgtc gtccgttttg gaacaccccg gttctggtgc acagcgtgag cccgcgtggt
     1321 aacaaggttc tgaccaaacg tagcagcatg agcctgccga cccaagtgag cttctataag
     1381 ggcggtgaca ttcaaagcgt tctgagcggc aaatttagcg cgtggctgat gggttacgac
     1441 ccggagtacc tggattatct ggttaagtat gaattccagc atgatgaagc agtggataac
     1501 aagtttaaca aagaacagca gaacgcgttt tatgagatct tgcacttacc gaatctgaac
     1561 gaggaacagc gcaatgcgtt cattcagtcc cttaaagacg atccatctca aagcgcaaat
     1621 ctgctggcag aagccaagaa actgaacgat gctcaagctc cgaaagtcga caacaaattc
     1681 aacaaagagc aacagaatgc gttctacgaa attctgcatt taccgaacct gaatgaagaa
     1741 cagcgtaacg cctttatcca atcgctgaaa gatgatccta gtcagtcagc caatctgttg
     1801 gcggaagcga agaaactcaa tgacgcccaa gctcccaaag ttgacgcgaa tagctggagc
     1861 catccgcagt ttgaaaagct ttagggctgt tttggcggat gagagaagat tttcagcctg
     1921 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
     1981 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     2041 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     2101 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     2161 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     2221 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     2281 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
     2341 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2401 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2461 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2521 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2581 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2641 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2701 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2761 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2821 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2881 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2941 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3001 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3061 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3121 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3181 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3241 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3301 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3361 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     3421 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3481 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3541 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3601 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3661 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3721 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3781 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3841 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3901 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3961 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     4021 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4081 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4141 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4201 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     4261 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     4321 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     4381 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4441 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc agatcaattc
     4501 gcgcgcgaag gcgaagcggc atgcataatg tgcctgtcaa atggacgaag cagggattct
     4561 gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa
     4621 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     4681 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     4741 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     4801 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa aatgtgacag
     4861 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     4921 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     4981 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     5041 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     5101 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     5161 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     5221 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     5281 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     5341 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     5401 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     5461 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagag
//
