LOCUS       sequence-203155-        6070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses murine Alkbh4 containing an N-ternimal 3xFlag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6070)
  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 6070)
  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-Alkbh4
FEATURES             Location/Qualifiers
     source          1..6070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    101..563
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(157..174)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     384..403
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             599..1402
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(662..681)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1272..1291
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1434..1451)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1440..1664
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1711..1730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1790..1806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1790..1806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1803..1825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1814..1830
                     /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(1838..1844,1845..1862,1863..1868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1883..1904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2021..2038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2192..2780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2272..2291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2951..3742,3743..3811))
                     /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     3574..3593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3812..3916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3991..4010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4182..4561
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4562..4765
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4810..4829
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4810..4828
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4862..4913
                     /label=CMVd2 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields moderately reduced expression 
                     relative to the full-length promoter"
     CDS             4943..5008
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5669..5695
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          5766..5995
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5978..5997
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
       61 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
      121 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
      181 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
      241 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
      301 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
      361 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
      421 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
      481 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
      541 tcctttgaaa aacacgatga taagcttgcc acaacccggg ataattcctg cagccaatat
      601 gggatcggcc attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
      661 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
      721 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
      781 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
      841 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
      901 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
      961 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     1021 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     1081 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     1141 gcccgacggc gatgatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     1201 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     1261 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     1321 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     1381 ccttcttgac gagttcttct gaggggatca attctctaga gctcgctgat cagcctcgac
     1441 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1501 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1561 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1621 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     1681 aaccagctgg ggctcgagtg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1741 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     1801 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1861 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1921 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1981 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2041 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2101 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2161 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2221 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2281 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2341 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2401 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2461 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2521 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2581 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2641 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2701 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2761 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2821 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2881 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2941 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3001 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3061 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3121 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3181 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3241 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3301 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3361 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3421 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3481 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3541 gaccgagttg ctcttgtccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3601 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3661 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3721 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3781 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3841 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3901 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     3961 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4021 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     4081 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     4141 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     4201 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     4261 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     4321 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     4381 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4441 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     4501 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4561 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     4621 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4681 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     4741 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
     4801 tatcgaaatt aatacgactc actataggga gacccaagct tggtaccgag ctcggatcga
     4861 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
     4921 tgaaccgtca gaattaacca tggactacaa agaccatgac ggtgattata aagatcatga
     4981 catcgattac aaggatgacg atgacaagct tgcggccgcg aattctgtgc gtctgatgga
     5041 ctgcgacccc tggaagctct cacagtctgg gaggaagaag caggactatg gccctaaagt
     5101 taattttcgg aagcagaagc tgaagatggc aggcttccag ggtctccctg gctttagcca
     5161 gaaggtagtc cagagaatgg gcctttaccc ggggctggag gacttccagc ctgtggaaca
     5221 gtgcaatctg gactacagcc ctgagcgggg ctccgccatc gacccccact tggacgacgc
     5281 ctggctgtgg ggcgagcggc tggtgagcct caacctcctg tcagccacag tggtgtccat
     5341 gtctccagag gcccctggca gcctgcttct ctgctcagcc ccctccgtca gacctgatgc
     5401 ttttgaggac agccttgtgg ctcctagcag gtctgtcccc tgccaggagg tggaggtggc
     5461 catcacagta ccccgccgct ccttgctggt gcttacaggg gccgcgaggc accagtggac
     5521 acacgccatc caccgcagac acatcaaggc ccgccgtgtg tgcgccacct tcagggagct
     5581 gtccagtgag ttcctgcctg gagggaagca gcaagaactg ggccaagaac tgctgcaggc
     5641 tgcattctcg ttccagggga gaccggtgta cccatacgat gttccagatt acgcttgagg
     5701 atccactagt aacggccgcc agtgtgctgg aattaattcg ctgtctgcga gggccagctg
     5761 ttggggtgag tactccctct caaaagcggg catgacttct gcgctaagat tgtcagtttc
     5821 caaaaacgag gaggatttga tattcacctg gcccgcggtg atgcctttga gggtggccgc
     5881 gtccatctgg tcagaaaaga caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga
     5941 gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc cacaggtgtc
     6001 cactcccagg tccaactgca ggtcgagcat gcatctaggg cggccaattc cgcccctctc
     6061 cccccccccc
//
