LOCUS       sequence-427145-        6265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG tagged mutant (UAF1 S170Y) UAF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6265)
  AUTHORS   Ng VH, Spencer Z, Neitzel LR, Nayak A, Loberg MA, Shen C, Kassel SN,
            Kroh HK, An Z, Anthony CC, Bryant JM, Lawson A, Goldsmith L, 
            Benchabane H, Hansen AG, Li J, D'Souza S, Lebensohn AM, Rohatgi R, 
            Weiss WA, Weiss VL, Williams C, Hong CC, Robbins DJ, Ahmed Y, Lee E
  TITLE     The USP46 complex deubiquitylates LRP6 to promote Wnt/beta-catenin 
            signaling.
  JOURNAL   Nat Commun. 2023 Oct 5;14(1):6173. doi: 10.1038/s41467-023-41836-z.
  PUBMED    37798301
REFERENCE   2  (bases 1 to 6265)
  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: "Nat 
            Commun."; date: "2023-10-5"; volume: "14(1):6173. doi"; pages: " 
            10.1038/s41467-023-41836-z"
COMMENT     Sequence Label: Human FLAG-UAF1 S170Y
FEATURES             Location/Qualifiers
     source          1..6265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      128..583
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(215..234)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     425..446
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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     complement(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /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)."
     primer_bind     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2754..2774
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    2887..3021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2924..2943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2978..2997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    3066..3090
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5136..5159)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(5206..5230)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(5285..5303)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5286..5303)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(join(5338..6265,1..57))
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 atccaccata ttgaattggc atgagtccat atacgccata ttgaattggc ggtcgactgg
       61 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
      121 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      181 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
      241 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
      301 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
      361 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
      421 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      481 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
      541 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc tcccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     2761 accctcacta aagggaacaa aagctgggta ccgggcccaa tgcattggcg ccgcggccgc
     2821 gaattaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     2881 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2941 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3001 gtatcttatc atgtctggat ctacgtaata cgactcacta tagttctaga ggctcgagag
     3061 gatcaaccac tttgtacaag aaagctgggt ctagatatct cgagtcacgt ggacttctga
     3121 cggtaatgga gggtgaggtc tccaccgctc ttccatatga agtgtttcac tgttcgaagg
     3181 tccatatttg gatccaaaac ctggtcctgg cacaaaagtt caattttctc ctctgccaac
     3241 acagcaatat cttcttcttt ttcctgttct cctggttttt cattattaga agagctagtg
     3301 gtttgagact cattatccaa gttgataatt ttttcataaa catgctccat aacttttcgg
     3361 acttggagca tgtcactagc agagagtcta tcttttttta aggtttttgc tcctgaagat
     3421 gcatgaggtt ggaggtagaa aggaattttg ttgaatttgg gcatattttt atccacagtg
     3481 atgtcaatta cccattgtgg cactgtttca ttaagaagca tagactcagt ctcacccccg
     3541 gaatctcggc agagcagcct gaacagtgtg cgacctccag cttcaccaaa gatcacgggt
     3601 gtatgtgggg gcacttgaaa atatccattt cccttctgca ctcggttctc ctgctcccca
     3661 tttacatggt ttacttcatt ttcctcttca tccattggat tcacatgtgt tctaggccaa
     3721 tattccagga gtgcttggag taaaagtcct cctaaattca attttggatc tgacccatca
     3781 gggctgctga aaccagcatc ttttgcagaa acccaggcag caaaacaatc actttcatcc
     3841 aaagtaatag ttaacatccc tgtttttaag tctactgaga accaatttgg cacatacacc
     3901 attttaaatc ttttcttaat ttcatcttca aaatccactt tgcccagatc ttcaacttta
     3961 catgccttca atacatccca atatgccaca ttattattgg tatctttggt taatatatgt
     4021 ctcttatcat taaaaatgtg gcactgaata atactagcac cccctttaat aacctggtca
     4081 ggttgtgtac aaagaggtgt tataggattt gtacagtcat tgtcataatc tccagaggct
     4141 ctaaaattat gaattccttt caaagtccat ttatttactg tagacttagt tgttgcaacc
     4201 caaattgcag gaggaggatc agctgatcta tcaagctcca tcttgagaac tggtgctttt
     4261 tcttcacaaa ttagcacccg aatgtcaggg tttcttaggt ctgtacaata aatcttcctg
     4321 tcccttccac cagaatacac atgtgtgaag gcatcattga cttgcagcgc ccaaacacct
     4381 tcatcatgga ctcggtatgt tgctatacat ctctgctggc caagggacca aaggcgaatt
     4441 gtcccatcag aactgcctga caggcattgc gtgccatctc tgtttaatag caatgccttc
     4501 acattatccg tgtgcccttt aagcttcatt agttttgcac atgttcttgg atcccataca
     4561 cgtaacacct tttcagtgga ccctgataca atgattgttc ccagttgatt catggccagg
     4621 ctataaatat aatctttgtt tccacttaaa gaagaagttg tgacagtgtt atttgaggca
     4681 gtcaatgctg ttagagtatt cacatcccaa aggaatattt gtctgtccaa cccagctgat
     4741 gctactagtt ctttatcctt ggcatatgct aaggccttta cgtaatcctt atgtgtcctt
     4801 aatgttgaca tgcaaaatcc cttgtgtgca ttccatactt ttactgtcgt gtcagaagaa
     4861 gcagatatta atgttttccc attacaacag agtacaatgt cgtttaccca atcagtatgg
     4921 tgttccatag atgctatata tggatcttgc ttgtgctgat tgacactcca tattcttatg
     4981 atagagtctc gaccggctgt gaaaagtcta tttagtgctg gatccagctg cagagcattg
     5041 actccatttc ggttgtactt ctccacttca tctcgaataa cataggaaac ctgcactttc
     5101 ctccgccctg ctgtgttctg ccggtgatgg gccgccttat cgtcgtcatc cttgtaatcg
     5161 cccatggtgg taccggatcc agtcgactga attggttccg gcgccagcct gcttttttgt
     5221 acaaacttgt tgatccttga attcgaatcg atgggatcct gcaaaaagaa caagtagctt
     5281 gtattctata gtgtcaccta aatcaagctt gctccgacgt ccccaggcag aatggcggtt
     5341 ccctaaacga gcattgctta tatagacctc ccattaggca cgcctaccgc ccatttacgt
     5401 caatggaacg cccatttgcg tcattgcccc tccccattga cgtcaatggg gatgtacttg
     5461 gcagccatcg cgggccattt accgccattg acgtcaatgg gagtactgcc aatgtaccct
     5521 ggcgtacttc caatagtaat gtacttgcca agttactatt aatagatatt gatgtactgc
     5581 caagtgggcc atttaccgtc attgacgtca atagggggcg tgagaacgga tatgaatggg
     5641 caatgagcca tcccattgac gtcaatggtg ggtggtccta ttgacgtcaa tgggcattga
     5701 gccaggcggg ccatttaccg taattgacgt caatggggga ggcgccatat acgtcaatag
     5761 gaccgcccat atgacgtcaa taggaaagac catatataga gaccattgac gtcaatgggg
     5821 gagtggctat gggcggtatt aggaagcccc atatatggta tatgggaccg cccattggga
     5881 ggggctatct acgtcaatag gaaaacccat atatggaata ctatatggca tagggccaat
     5941 acatagtatt gaacctggcc aatagccata ttggcatagg gccatattgg atattgccta
     6001 tatattgatc ctggcatata gccaatatgg ccgccattat tggcaccatg ccaattcaat
     6061 atggcggact tggcacaggg ccaagacccc tcctcaaact tggcaccgtg ccaagtagac
     6121 ccctccccag ttggcacagt gccaggtccg ccatattgaa ttggcatggt gccaagtccg
     6181 ccatattgaa ttggctgggg tccagatcca ccatattgaa ttggcacgag tccatatacg
     6241 ccatattgaa ttggcacagg tccag
//
