LOCUS       sequence-390276-        5570 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express NP_049039 in bacterial cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5570)
  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 5570)
  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-NP_049039
FEATURES             Location/Qualifiers
     source          1..5570
                     /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             1909..1932
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1971..1988)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2141..2227
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2319..2346
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2365..2456
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2457..2525,2526..3317)
                     /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(2675..2694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3488..4076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3977..3996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4230..4247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4262..4402
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4328..4347)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4488..4510
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4666..5544)
                     /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 tccatgtcat ttaggacatt agaactattc gctggcatcg ccggtatcag ccacggcctg
      481 cgtggtattt ccaccccggt tgccttcgtg gaaatcaacg aagatgcgca aaaattcctg
      541 aaaacgaagt tctcagacgc gagcgtgttc aacgacgtga ccaaatttac gaagagcgac
      601 ttcccggagg acatcgatat gattactgcg ggttttccgt gtaccggctt ctcgatcgct
      661 ggcagccgta ccggcttcga gcacaaagag tccggtctgt ttgctgatgt ggtgcgtatt
      721 accgaggaat ataaaccgaa gatcgtgttt ttggagaact cgcacatgct gagccacacc
      781 tacaatctgg atgtagtcgt caaaaagatg gatgaaattg gttacttctg caagtgggtt
      841 acctgtcgtg cgagcattgt tggtgcacat catcagcgtc accgctggtt ttgcctggcg
      901 atccgcaagg actatgaacc gaaagagatc atcgttagcg tgaatgccac caagttcgat
      961 tgggaaaaca acgagccgcc ttgcaaggtt gataataaat cttacgagaa cagcacattg
     1021 gtacgcctgg ctggctatag cgtggttccg gatcaaattc gttatgcgtt taccggcctc
     1081 ttcaccggtg attttgaatc cagctggaaa acgactttaa ccccaggtgc gattaacggt
     1141 actgagcaca agaagatgaa aggtacgtac gacaaggtga ttaacggcta ctacgagaac
     1201 gacgtgtatt actccttttc ccgcaaagag gttcatagag caccgctgaa tatttctgtc
     1261 aagccgcgtg acattccgga aaaacacaac ggtaaaactt tggtcgaccg tgaaatgatc
     1321 aaaaagtatt ggtgcacccc gtgcgcgagc tacggcacgg ctaccgcggg ttgcaatgtt
     1381 ctgaccgatc gtcagagcca tgcactgccg acccaggttc gtttttctta ccgcggagtt
     1441 tgtggtcgtc atctgagcgg catctggtgc gcatggctga tgggttacga ccaagaatac
     1501 ctaagctatt tggtgcagta tgacgaattc cagcatgatg aagcagtgga taacaagttt
     1561 aacaaagaac agcagaacgc gttttatgag atcttgcact taccgaatct gaacgaggaa
     1621 cagcgcaatg cgttcattca gtcccttaaa gacgatccat ctcaaagcgc aaatctgctg
     1681 gcagaagcca agaaactgaa cgatgctcaa gctccgaaag tcgacaacaa attcaacaaa
     1741 gagcaacaga atgcgttcta cgaaattctg catttaccga acctgaatga agaacagcgt
     1801 aacgccttta tccaatcgct gaaagatgat cctagtcagt cagccaatct gttggcggaa
     1861 gcgaagaaac tcaatgacgc ccaagctccc aaagttgacg cgaatagctg gagccatccg
     1921 cagtttgaaa agctttaggg ctgttttggc ggatgagaga agattttcag cctgatacag
     1981 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg
     2041 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     2101 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     2161 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
     2221 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc
     2281 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
     2341 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
     2401 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2461 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2521 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2581 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2641 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     2701 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2761 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2821 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2881 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2941 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3001 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3061 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3121 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3181 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3241 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3301 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3361 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3421 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3481 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3541 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3601 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3661 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3721 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3781 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3841 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3901 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3961 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4021 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4081 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4141 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4201 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4261 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     4321 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     4381 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     4441 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4501 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     4561 gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
     4621 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
     4681 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
     4741 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
     4801 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
     4861 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaaaatgtg acagacgcga
     4921 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
     4981 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
     5041 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
     5101 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
     5161 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     5221 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     5281 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     5341 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
     5401 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     5461 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     5521 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag
//
