LOCUS       sequence-254501-        6073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses an N-terminally truncated version of FLAG-tagged mouse 
            TMEM98 in mammalian cells (C-terminal tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6073)
  AUTHORS   van der Wal T, Lambooij JP, van Amerongen R
  TITLE     TMEM98 is a negative regulator of FRAT mediated Wnt/ss-catenin 
            signalling.
  JOURNAL   PLoS One. 2020 Jan 21;15(1):e0227435. doi: 
            10.1371/journal.pone.0227435. eCollection 2020.
  PUBMED    31961879
REFERENCE   2  (bases 1 to 6073)
  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: "PLoS One.";
            date: "2020-01-21"; volume: "15(1):e0227435. doi"; pages: " 
            10.1371/journal.pone.0227435. eCollection 2020"
COMMENT     Sequence Label: pCDNA3.1-Tmem98deltaN-FLAG
FEATURES             Location/Qualifiers
     source          1..6073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1492..1515
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1626..1644)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1627..1644)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1664..1681)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1670..1894
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1940..2368
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2027..2046)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2237..2258
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2377..2397)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2382..2711
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2562..2697
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2624..2643
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2778..3572
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2832..2851)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3442..3461
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3746..3867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3783..3802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3837..3856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3916..3932)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3916..3932)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3929..3951)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3940..3956
                     /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(3964..3970,3971..3988,3989..3994))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4009..4030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4147..4164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4318..4906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4398..4417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5077..5868,5869..5937))
                     /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     5700..5719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5938..6042)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccatg
      901 gcccctcgag acctactaca gcgttatgat tccaagccca tcgtggacct cattggtgct
      961 atggagacgc agtctgaacc ctccgagctg gagctggatg atgtcgtcat caccaacccc
     1021 cacatcgagg ccatcctgga gaatgaggac tggatcgaag atgcctcggg cctcatgtcc
     1081 cactgcatcg ccatcttgaa gatttgtcac actctgacag aaaaactcgt tgccatgaca
     1141 atgggttctg gggccaagat gaagacgtca gcaagtgtca gtgacatcat tgtggtggcc
     1201 aaacggatta gccccagagt ggacgacgtc gtgaagtcaa tgtaccctcc actggacccc
     1261 aagctcctgg atgcacggac aaccgccctg ctgctgtccg ttagtcactt ggtgctagtg
     1321 accaggaacg cctgccatct aaccgggggc ctggactgga ttgaccaatc actgtctgcc
     1381 gctgaggagc acctggaagt ccttcgagag gcagccctgg cttctgagcc agataaaagc
     1441 ctccccaacc ctgagggctt cctgcaggaa cagtcggcca ttgcaaccat ggactacaaa
     1501 gacgatgacg acaagcttgg taccgagctc ggatcgatcc actagtaacg gccgccagtg
     1561 tgctggaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc
     1621 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     1681 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1741 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1801 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1861 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     1921 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1981 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2041 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2101 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2161 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2221 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2281 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2341 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2401 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2461 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2521 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2581 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2641 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2701 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     2761 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2821 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2881 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2941 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3001 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3061 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3121 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3181 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3241 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3301 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3361 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3421 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3481 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3541 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3601 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3661 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3721 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3781 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3841 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     3901 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3961 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4021 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4081 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4141 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4201 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4261 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4321 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4381 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4441 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4501 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4561 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4621 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4681 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4741 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4801 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4861 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4921 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4981 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5041 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5101 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5161 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5221 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5281 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5341 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5401 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5461 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5521 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5581 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5641 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5701 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5761 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5821 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5881 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6001 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6061 gccacctgac gtc
//
