LOCUS       sequence-321754-        3314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Site-specific DNA methyltransferase 2; protects DNA from cleavage by
            Eco31I.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3314)
  TITLE     Open Enzyme Collection
REFERENCE   2  (bases 1 to 3314)
  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"
COMMENT     Sequence Label: pOpen-Eco31IB
FEATURES             Location/Qualifiers
     source          1..3314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..42)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      38..85
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(90..106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(90..106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(173..761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(253..272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(952..1743,1744..1812))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1575..1594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1813..1917)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1933..1949
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1950..1981
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     RBS             2602..2610
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 cgctgaggtg tcaatcgtcg gagccgctga gcaataacta gcataacccc ttggggcctc
       61 taaacgggtc ttgaggggtt ttttgcatgg tcatagctgt ttcctgagag cttggcaggt
      121 gatgacacac attaacaaat ttcgtgagga gtctccagaa gaatgccatt aatttccata
      181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
      721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaaggccta ctattagcaa
      781 caacgatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      841 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      901 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      961 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1021 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1081 aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc
     1141 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1201 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1261 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1321 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1381 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1441 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1501 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1561 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     1621 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     1681 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1741 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1801 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1861 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1921 ctgcaccagt cagtaaaacg acggccagta gtcaaaagcc tccgaccgga ggcttttgac
     1981 ttggttcagg tggagtggga gaaataagac gacgaccctc gggtctctaa tgactaaatc
     2041 tgagacattc atgattccaa accacaaagc ggcgaagctg agcgaattag atatgatgat
     2101 tgtgaactct gttccgccgg gtggtaattg gaaaaacatc cctctggacg tcccgtcaaa
     2161 acgtattgaa caaatacgag atagttatgc tcagggaaaa ggcagccgca gcacttatta
     2221 tggcagactc ctccccgata tgcccgctta caccattaat acgtatttta accgccccgg
     2281 caacggctgc catatccatt acgaacagga tcgtgtactg agccagcgtg aagcggcccg
     2341 gcttcaaagc ttccctgatg acttcatctt tttcggtggc cagacagcga ttaacaccca
     2401 gatcggcaat gctgtaccgc cgtttctggc atttctgata gcaaaagaga ttgaaaaagc
     2461 catcggcaac acgggctatt atattgacct gttttccggt gcagggggtt taggtctggg
     2521 atttaagtgg gccggctgga ccccgttact ggcgaacgat atcgaggaga aataccttca
     2581 aacgtattca aataacgtgc ataaggaggt gctgtgtggg agtatcagtg acaacgaaac
     2641 cttttccaaa atcgcggata aaatttctgg gtttaaaaaa ttgtacttcg acaagcaact
     2701 atggattctg ggcggtccac cgtgccaggg tttttcgact gcggggaatg cccgtacgat
     2761 ggacgatcct cgtaattcgc tgttcatgca ttacaagagt ctcttgaacg aaatcaaacc
     2821 gaatggcttc attttcgaaa atgttgcagg attgttaaat atggaaaaag gtaaagtttt
     2881 tgagcgcgtg aaagaagaat tttcgtccac gatgaaaacc atgaacggct ggatactgaa
     2941 ctcggaacac tacgccattc cgcagcgccg caaacgtgtg attctagtcg gaagcaatga
     3001 tccattgttt tcaatcgaac cgccgcagaa gctgaccgag gataaagagt catgggtcag
     3061 tgttaaagat gcactgtctg atcttccacc tcttcaacac ggggaagatg gtagcggcaa
     3121 atattacatt catcacccag aaaatgacta tcagctgttc atgcggggta acattacccc
     3181 gtccgaatac tatgaacgca atatcaagcc gagcctgtga agagctttga gaccggcctt
     3241 ttattgggtc cagtccgcaa accctgatct tattttaggg ataacagggt aatgtacaaa
     3301 tagggtcgaa attc
//
