LOCUS       sequence-426357-        4965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dTAG for C-terminal tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4965)
  AUTHORS   Hatoyama Y, Islam M, Bond AG, Hayashi KI, Ciulli A, Kanemaki MT
  TITLE     Combination of AID2 and BromoTag expands the utility of degron-based
            protein knockdowns.
  JOURNAL   EMBO Rep. 2024 Sep;25(9):4062-4077. doi: 10.1038/s44319-024-00224-4.
            Epub 2024 Aug 23.
  PUBMED    39179892
REFERENCE   2  (bases 1 to 4965)
  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: 
            "10.1038/s44319-024-00224-4"; journalName: "EMBO Rep"; date: 
            "2024-09"; volume: "25"; issue: "9"; pages: "4062-4077"
COMMENT     Sequence Label: pMK454 (dTAG-BSD)
FEATURES             Location/Qualifiers
     source          1..4965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             681..1004
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    1019..1140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1056..1075)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1110..1129
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1171..1204
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1265..1764
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1300..1323)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1697..1716
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1788..1797
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1794..2192
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    2695..2728
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2777..2797)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2777..2795)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2816..2832)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2816..2832)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2829..2851)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2840..2856
                     /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(2864..2870,2871..2888,2889..2894))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2909..2930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3047..3064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3218..3806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3298..3317)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3977..4768,4769..4837))
                     /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     4600..4619
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4838..4942)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctccc
      661 cgggatccgg tgcaggcgcc ggagtgcagg tggaaaccat ctccccagga gacgggcgca
      721 ccttccccaa gcgcggccag acctgcgtgg tgcactacac cgggatgctt gaagatggaa
      781 agaaagttga ttcctcccgg gacagaaaca agccctttaa gtttatgcta ggcaagcagg
      841 aggtgatccg aggctgggaa gaaggggttg cccagatgag tgtgggtcag agagccaaac
      901 tgactatatc tccagattat gcctatggtg ccactgggca cccaggcatc atcccaccac
      961 atgccactct cgtcttcgat gtggagcttc taaaactgga ataacaattg ttgttgttaa
     1021 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1081 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1141 acgcgtcgat catattcaat aacccttaat ataacttcgt ataatgtatg ctatacgaag
     1201 ttattaggtc tgaagaggag tttacgtcca gccaagctta ggatctcgac ctcgaaattc
     1261 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     1321 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     1381 aggcgccaac cgactccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     1441 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     1501 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     1561 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     1621 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     1681 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     1741 tcttcctcat ctccgggcct ttcgacctgc atccatctag atctcgagcc accatggcca
     1801 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     1861 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     1921 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     1981 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     2041 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     2101 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     2161 tgctgccctc tggttatgtg tgggagggct aatcgatgat atcagatccc cgggatgcag
     2221 aaattgatga tctattaaac aataaagatg tccactaaaa tggaagtttt tcctgtcata
     2281 ctttgttaag aagggtgaga acagagtacc tacattttga atggaaggat tggagctacg
     2341 ggggtggggg tggggtggga ttagataaat gcctgctctt tactgaaggc tctttactat
     2401 tgctttatga taatgtttca tagttggata tcataattta aacaagcaaa accaaattaa
     2461 gggccagctc attcctccca ctcatgatct atagatctat agatctctcg tgggatcatt
     2521 gtttttctct tgattcccac tttgtggttc taagtactgt ggtttccaaa tgtgtcagtt
     2581 tcatagcctg aagaacgaga tcagcagcct ctgttccaca tacacttcat tctcagtatt
     2641 gttttgccaa gttctaattc catcagaagc tggtcgagat ccggaaccct taatataact
     2701 tcgtataatg tatgctatac gaagttatta ggtccctcga agaggttcac taggatccgg
     2761 tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     2821 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2881 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2941 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3001 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3061 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3121 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3181 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3241 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3301 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3361 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3421 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3481 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3541 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3601 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3661 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3721 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3781 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3841 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3901 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3961 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4021 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4081 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4141 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4201 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4261 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4321 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4381 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4441 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4501 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4561 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4621 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4681 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4741 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4801 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4861 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4921 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
