LOCUS       sequence-13765-e        2830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2830)
  TITLE     latest plasmids
REFERENCE   2  (bases 1 to 2830)
  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: pENTR4-CLIPf (w877-2)
FEATURES             Location/Qualifiers
     source          1..2830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             463..1008
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylcytosine 
                     derivatives"
                     /label=CLIP-tag(TM)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLIQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISESHLAALVGNPAATAAVNTALDGNPVPILIPCHRVVQGDSDVGPYLGGLA
                     VKEWLLAHEGHRLGKPGLG"
     protein_bind    complement(1059..1158)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1270..1289)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1281..2090
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1348..1367)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2180..2768
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2669..2688
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatggacaa agactgcgaa
      481 atgaagcgca ccaccctgga tagccctctg ggcaagctgg aactgtctgg gtgcgaacag
      541 ggcctgcacc gtatcatctt cctgggcaaa ggaacatctg ccgccgacgc cgtggaagtg
      601 cctgccccag ccgccgtgct gggcggacca gagccactga tccaggccac cgcctggctc
      661 aacgcctact ttcaccagcc tgaggccatc gaggagttcc ctgtgccagc cctgcaccac
      721 ccagtgttcc agcaggagag ctttacccgc caggtgctgt ggaaactgct gaaagtggtg
      781 aagttcggag aggtcatcag cgagagccac ctggccgccc tggtgggcaa tcccgccgcc
      841 accgccgccg tgaacaccgc cctggacgga aatcccgtgc ccattctgat cccctgccac
      901 cgggtggtgc agggcgacag cgacgtgggg ccctacctgg gcgggctcgc cgtgaaagag
      961 tggctgctgg cccacgaggg ccacagactg ggcaagcctg ggctgggtcc tgcaggcgga
     1021 tccggtaccg aattcgcggc cgcactcgag atatctagac ccagctttct tgtacaaagt
     1081 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1141 aaataaaatc attatttgcc atccagctgc agctctggcc cgtgtctcaa aatctctgat
     1201 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     1261 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     1321 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     1381 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     1441 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     1501 cggaatttat gcctcttccg accatcaagc attttatccg tactcctggt gatgcatggt
     1561 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     1621 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     1681 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     1741 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     1801 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     1861 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     1921 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     1981 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     2041 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     2101 ttaattggtt gtaacattat tcagattggg ccccgttcca ctgagcgtca gaccccgtag
     2161 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2221 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2281 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2341 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2401 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2461 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2521 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2581 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2641 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2701 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2761 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2821 ctcacatgtt
//
