LOCUS       sequence-389729-        5519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express YP_001497877 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5519)
  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 5519)
  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_001497877
FEATURES             Location/Qualifiers
     source          1..5519
                     /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             1858..1881
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1920..1937)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2090..2176
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2268..2295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2314..2405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2406..2474,2475..3266)
                     /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(2624..2643)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3437..4025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3926..3945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4179..4196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4211..4351
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4277..4296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4437..4459
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4615..5493)
                     /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 tccatgagcc tgaaagcgct ggagctgttc gcgggtatcg gtggcattac ctacggcctg
      481 cgtggttatg tggaaccgat cgcgttctgc gagtacgaaa aagatgcggc ggcgtttctg
      541 agccagcgtg gcctgccggt tcacggtgac attaccaagt ttgatgcgag cgtgtataag
      601 accaaaatcg acattgttac cgcgggttgg ccgtgcaccg gtttcagcac cgcgggcaag
      661 ggcaccggtt ttgatcacga ggcgagcggt ctgtggaccg aagtgatccg tgtggttaag
      721 gagagcgaac cgaaatacgt gttcctggag aacagccacg ttctggcgca aaccaagaac
      781 ctgaaagtta tcattcacga cctggatggc ctggaatacg acacccgttg gtggacctgc
      841 cgtagcaacg atgtgaacgt tggcgcgcac cacaaccgtt atcgttggtt catgctggcg
      901 gagaagaaag gtagcgtgac caagttcgat aagatccagg ttgaaaagtt catctggagc
      961 ggcgacttta aggataaaca agtgagcgag aacagccacg aaaacaagca actgatcaaa
     1021 ttcatgggta acagcgtggt tccggaccaa gtgcgttacg cgtttgagag catgagcgat
     1081 atgaaactgg aaggcaccct ggttaacgac aaggatgaga ttgtgaaagt tggctacagc
     1141 aaggacggta tcatgtataa gattcacgtg gatcacaaga tcattccgaa actgaacatt
     1201 gttctgaccc cgcgtgaccc gccggagggt cacaaagcgc gtgaagatgc gatcattaag
     1261 agcccgattc tgatgaccta ctggaacacc ccggcgtttt gctatcacaa aagcgcgcgt
     1321 ggcagcaagc tgctgaccaa gcgtcagaaa aacaacctgc acacccaaat caagttctgc
     1381 ccgggtggca gcgacgatgg ctacctgagc ggtcgttttt gcgcgtggct gatgggctac
     1441 ggtgacgaat atctgggtaa cctgattgat tatgaattcc agcatgatga agcagtggat
     1501 aacaagttta acaaagaaca gcagaacgcg ttttatgaga tcttgcactt accgaatctg
     1561 aacgaggaac agcgcaatgc gttcattcag tcccttaaag acgatccatc tcaaagcgca
     1621 aatctgctgg cagaagccaa gaaactgaac gatgctcaag ctccgaaagt cgacaacaaa
     1681 ttcaacaaag agcaacagaa tgcgttctac gaaattctgc atttaccgaa cctgaatgaa
     1741 gaacagcgta acgcctttat ccaatcgctg aaagatgatc ctagtcagtc agccaatctg
     1801 ttggcggaag cgaagaaact caatgacgcc caagctccca aagttgacgc gaatagctgg
     1861 agccatccgc agtttgaaaa gctttagggc tgttttggcg gatgagagaa gattttcagc
     1921 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     1981 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     2041 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     2101 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     2161 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     2221 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     2281 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     2341 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2401 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2461 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2521 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2581 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2641 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2701 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2761 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2821 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2881 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     2941 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3001 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3061 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3121 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3181 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3241 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3301 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3361 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3421 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3481 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3541 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3601 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3661 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3721 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3781 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3841 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3901 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3961 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4021 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4081 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4141 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4201 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4261 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     4321 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     4381 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4441 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agcagatcaa
     4501 ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt caaatggacg aagcagggat
     4561 tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga
     4621 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct
     4681 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt
     4741 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat
     4801 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaaaatgtga
     4861 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc
     4921 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg
     4981 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc
     5041 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc
     5101 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga
     5161 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata
     5221 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa
     5281 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg
     5341 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag
     5401 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc
     5461 cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagag
//
