LOCUS       sequence-165735-        6091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YedK in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6091)
  AUTHORS   Kweon SM, Zhu B, Chen Y, Aravind L, Xu SY, Feldman DE
  TITLE     Erasure of Tet-Oxidized 5-Methylcytosine by a SRAP Nuclease.
  JOURNAL   Cell Rep. 2017 Oct 10;21(2):482-494. doi: 
            10.1016/j.celrep.2017.09.055.
  PUBMED    29020633
REFERENCE   2  (bases 1 to 6091)
  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"; doi: 
            "10.1016/j.celrep.2017.09"; journalName: "Cell Rep"; date: 
            "2017-10-10- 10"; volume: "21"; issue: "2"; pages: "482-494"
COMMENT     Sequence Label: p3XFlag-YedK
FEATURES             Location/Qualifiers
     source          1..6091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1663..1692
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             join(1702..1704,1705..1707,1708..2424)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKS"
     primer_bind     complement(1747..1768)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2008..2027)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2355..2376
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2432..3054
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2631..2650)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3078..3098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3083..3412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3263..3398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3450..3469)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3451..3469)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3483..3499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3483..3499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3496..3518)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3507..3523
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3531..3537,3538..3555,3556..3561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3576..3597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3714..3731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3885..4473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3965..3984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4644..5435,5436..5504))
                     /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     5267..5286
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5505..5609)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5636..6064
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5723..5742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5933..5954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttt gtggacgctt tgcccaatcc
     1021 caaacgcgtg aagattacct tgcgcttctc gcggaagata ttgaacgcga tattccctat
     1081 gatcccgaac ccattggcag atacaacgtc gcgccgggaa ccaaagtcct actgctcagt
     1141 gaacgtgatg aacaccttca tctggatccg gttttctggg gatatgctcc cggatggtgg
     1201 gataaaccgc cgctgattaa cgcccgcgta gaaactgcgg caaccagtcg tatgtttaaa
     1261 ccgctctggc aacatggtcg ggcaatctgt tttgccgatg gctggtttga gtggaaaaaa
     1321 gaaggcgaca aaaaacagcc tttttttatc tatcgtgctg atggacaacc tatttttatg
     1381 gccgcgatag gtagcacacc atttgagcgt ggtgacgaag ccgaaggatt tttgatcgtc
     1441 actgctgcgg cagatcaagg tctggtagat attcatgacc gccgcccact ggtactgtcg
     1501 ccagaagctg cgcgggaatg gatgcggcaa gagattagcg gtaaagaagc ctcagaaatc
     1561 gcggccagtg gctgtgttcc agcaaaccag ttctcctggc acccggtatc gcgcgcggtg
     1621 ggtaatgtta aaaaccaggg cgcggagtta attcaacctg ttgaacaaaa actcatctca
     1681 gaagaggatc tgtagaattc aatggtgagc aagggcgagg agctgttcac cggggtggtg
     1741 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1801 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1861 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     1921 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1981 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2041 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2101 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2161 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2221 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2281 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     2341 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2401 atggacgagc tgtacaagtc ctgaggatcc cgggtggcat ccctgtgacc cctccccagt
     2461 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     2521 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     2581 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     2641 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     2701 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     2761 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     2821 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     2881 gtgaaccact gctcccttcc ctgtccttct gattttaaaa taactatacc agcaggagga
     2941 cgtccagaca cagcataggc tacctggcca tgcccaaccg gtgggacatt tgagttgctt
     3001 gcttggcact gtcctctcat gcgttgggtc cactcagtag atgcctgttg aattgggtac
     3061 gcggccagct tggctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3241 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3301 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3361 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcct
     3421 cgaggaactg aaaaaccaga aagttaattc cctatagtga gtcgtattaa attcgtaatc
     3481 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3541 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3601 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3661 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3721 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3781 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3841 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3901 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3961 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4021 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4081 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4141 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4201 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4261 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4321 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4381 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4441 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4501 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4561 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4621 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4681 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4741 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4801 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4861 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4921 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4981 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5041 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5101 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5161 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5221 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     5281 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     5341 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     5401 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     5461 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5521 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5581 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     5641 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5701 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5761 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5821 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5881 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5941 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6001 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6061 aattttaaca aaatattaac gcttacaatt t
//
