LOCUS       sequence-235098-        6200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E. coli Dam methyltransferase containing an N-ternimal 
            3xFlag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6200)
  AUTHORS   Kweon SM, Chen Y, Moon E, Kvederaviciute K, Klimasauskas S, Feldman 
            DE
  TITLE     An Adversarial DNA N(6)-Methyladenine-Sensor Network Preserves 
            Polycomb Silencing.
  JOURNAL   Mol Cell. 2019 Apr 5. pii: S1097-2765(19)30222-9. doi: 
            10.1016/j.molcel.2019.03.018.
  PUBMED    30982744
REFERENCE   2  (bases 1 to 6200)
  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: "Mol Cell. 
            2019 Apr 5. pii: S1097-2765(19)30222-9. doi: 
            10.1016/j.molcel.2019.03.018."
COMMENT     Sequence Label: p3xFlag-Dam
FEATURES             Location/Qualifiers
     source          1..6200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        457..660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     611..631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             690..755
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(1601..1603,1604..1606,1607..2323)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPICDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKS"
     primer_bind     complement(1646..1667)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1907..1926)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2254..2275
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2331..2953
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2530..2549)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2977..2997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3318..3337)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3319..3337)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3351..3367)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3351..3367)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3364..3386)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3375..3391
                     /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(3399..3405,3406..3423,3424..3429))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3444..3465
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3582..3599)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3753..4341)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3833..3852)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4512..5303,5304..5372))
                     /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     5135..5154
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5373..5477)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5504..5959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5591..5610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5801..5822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6085..6107
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6099..6116
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6100..6116
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca
       61 atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      121 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      241 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      301 cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac
      361 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg
      421 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc
      481 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      541 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc
      601 gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      661 cgtttagtga accgtcagaa ttaaccatgg actacaaaga ccatgacggt gattataaag
      721 atcatgacat cgattacaag gatgacgatg acaagcttat gaagaagaat cgtgcttttt
      781 taaagtgggc cggcggcaag tatcccttat tagacgacat caagagacat ttacccaagg
      841 gcgagtgttt agtggagccc tttgtgggcg ccggaagcgt gtttttaaac accgacttct
      901 ctcgttacat tttagccgac atcaacagcg acctcatcag cctctacaat atcgtgaaga
      961 tgaggaccga cgaatacgtg caagctgcca gagagctgtt tgtgcccgaa accaactgcg
     1021 ccgaggtgta ttaccagttt cgtgaggagt tcaataagag ccaagatccc tttcgtaggg
     1081 ccgtgctgtt tttatattta aatcgttacg gctacaacgg tttatgtcgt tacaatctga
     1141 ggggcgagtt caacgtcccc ttcggcagat acaagaagcc ctacttcccc gaagccgaac
     1201 tgtaccactt cgccgagaag gcccagaacg ccttcttcta ctgcgagtcc tatgccgatt
     1261 ccatggccag agccgacgac gccagcgtcg tgtattgtga tcccccctac gctcctctga
     1321 gcgctaccgc caacttcacc gcctaccaca ccaacagctt cacactggag cagcaagctc
     1381 atttagccga gattgccgaa ggtttagtgg agaggcacat ccccgttctg atctccaacc
     1441 acgataccat gctgactcgt gagtggtacc agagagccaa gctgcacgtg gtgaaagtga
     1501 ggaggagcat ctcctccaac ggaggcacaa ggaagaaggt cgatgagctg ctggctttat
     1561 ataagcccgg cgtggtcagc cccgccaaga agtgaattca atggtgagca agggcgagga
     1621 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1681 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1741 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
     1801 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1861 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1921 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1981 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2041 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2101 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2161 ccccatctgc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccgc
     2221 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2281 cgccgggatc actctcggca tggacgagct gtacaagtcc tgaggatccc gggtggcatc
     2341 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     2401 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     2461 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     2521 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     2581 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     2641 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     2701 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     2761 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
     2821 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
     2881 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
     2941 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
     3001 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3061 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3121 tcaattagtc agcaaccata gtcccgcccc taactccgcc cattctccgc cccatggctg
     3181 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3241 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcctcg aggaactgaa
     3301 aaaccagaaa gttaattccc tatagtgagt cgtattaaat tcgtaatcat gtcatagctg
     3361 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3421 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3481 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3541 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3601 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3661 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3721 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3781 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3841 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3901 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     3961 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4021 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4081 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4141 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4201 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4261 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4321 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4381 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4441 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4501 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4561 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4621 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4681 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4741 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4801 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4861 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4921 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4981 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5041 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5101 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5161 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5221 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5281 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5341 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5401 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5461 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
     5521 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5581 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5641 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5701 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5761 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5821 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5881 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5941 atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6001 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6061 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     6121 agctgatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     6181 cataaatcaa tattggctat
//
