LOCUS       sequence-390264-        5579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YP_001498439 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5579)
  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 5579)
  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_001498439
FEATURES             Location/Qualifiers
     source          1..5579
                     /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             1297..1308
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     CDS             1918..1941
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1980..1997)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2150..2236
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2328..2355
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2374..2465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2466..2534,2535..3326)
                     /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(2684..2703)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3497..4085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3986..4005
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4239..4256
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4271..4411
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4337..4356)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4497..4519
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4675..5553)
                     /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 tccatgttag atacactaga attgtttgct ggaatcggcg gcattaccta tggtctgcgt
      481 ggtttcgcga agccggccgc atttgttgag tggaacgaag aagcgaagaa cgtgcttaag
      541 cgccacaaag ttccgatctt tgatgatgtt actacgtttg acgctacccc gtttaaagac
      601 aaagttgaca tggtttctgc aggctggccg tgtaccggtt tcagcaccgc gggtcacggc
      661 accggcttct cccatgaggc cagcggcctg tttgtggagg ttgtccgtgt tatcaaagaa
      721 tgccagccga attttgtttt tctggaaaac agccacgttc tcgcgcaaac ccgtttccta
      781 aaggtggtgc tgagcagtct ggatgaactg ggttacgacg cgcgttggat gagctgtaaa
      841 tcaacctgtg ttggcgcaat tcatgagcgc catcgctggt tttgcctggc taccaaaaga
      901 ggttttgtgc caccaacatt gacgagccag gtcaagcacg aacgtttcga ctgggattcg
      961 gatgaaccgg ctaagcaagt tccgaaaaac accgaggaga acaagatgat gatccgtctg
     1021 gcgggcaact ccgtggtgcc agaccagatt cgtttcgtgt tcaaaaaact gtacagcggt
     1081 ttctcttcga ccgttgacac cacttccaat gatgtagaga tcatccattg gaatccgctg
     1141 ctggaaaaca acggtttcgt gggtaatatt caccaggtca agaaacagcg tattagcaat
     1201 ggcttctgca ttgacggccg tatccacgag aaacacgtgg tcatcacgca tcgtaatccg
     1261 atcaacaaaa ccttgtaccc gaacccgctg ccggataatc acaaagtgaa ggacttgaag
     1321 aacgttgtaa ccaataacgt gacgaagaga ttctggtcca ctccgtgcca taccgatttt
     1381 cgccgtacct ctccggttgt cttaactaag cgcttgttga cgaacctgcc tgcgcaagta
     1441 cgcttcgtcg aggactccgt gccgggttgg cgtttgtctg gtgaatggtg cctgtggctg
     1501 atgggttata acaagagcta tgttaccggt agcgaattcc agcatgatga agcagtggat
     1561 aacaagttta acaaagaaca gcagaacgcg ttttatgaga tcttgcactt accgaatctg
     1621 aacgaggaac agcgcaatgc gttcattcag tcccttaaag acgatccatc tcaaagcgca
     1681 aatctgctgg cagaagccaa gaaactgaac gatgctcaag ctccgaaagt cgacaacaaa
     1741 ttcaacaaag agcaacagaa tgcgttctac gaaattctgc atttaccgaa cctgaatgaa
     1801 gaacagcgta acgcctttat ccaatcgctg aaagatgatc ctagtcagtc agccaatctg
     1861 ttggcggaag cgaagaaact caatgacgcc caagctccca aagttgacgc gaatagctgg
     1921 agccatccgc agtttgaaaa gctttagggc tgttttggcg gatgagagaa gattttcagc
     1981 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     2041 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     2101 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     2161 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     2221 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     2281 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     2341 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     2401 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2461 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2521 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2581 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2641 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2701 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2761 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2821 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2881 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2941 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3001 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3061 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3121 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3181 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3241 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3301 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3361 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3421 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3481 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3541 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3601 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3661 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3721 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3781 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3841 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3901 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3961 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4021 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4081 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4141 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4201 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4261 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4321 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     4381 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     4441 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4501 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa
     4561 ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt caaatggacg aagcagggat
     4621 tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga
     4681 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct
     4741 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt
     4801 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat
     4861 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaaaatgtga
     4921 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc
     4981 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg
     5041 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc
     5101 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc
     5161 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga
     5221 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata
     5281 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa
     5341 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg
     5401 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag
     5461 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc
     5521 cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagag
//
