LOCUS       sequence-254378-        5421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mTurquoise2-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 5421)
  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 5421)
  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: Tmem98-mTq2
FEATURES             Location/Qualifiers
     source          1..5421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1362..1371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1368..1370,1371..1373,1374..2087)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1413..1434)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1674..1693)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2021..2042
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2209..2330
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2246..2265)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2300..2319
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2337..2792)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2474..2495)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2686..2705
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2819..2923
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2925..3282
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2925..3116
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2948..2968)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3133..3268
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3317..4111
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3317..4111
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3371..3390)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3981..4000
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4299..4318)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4343..4390
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4719..5307
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5208..5227
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccca
      661 tggagactgt ggtgatcgtc gccataggtg tgctggccac cattttcctg gcctcatttg
      721 ctgctttggt ggtggtctgt aggcagcgtt actgccggcc tcgagaccta ctacagcgtt
      781 atgattccaa gcccatcgtg gacctcattg gtgctatgga gacgcagtct gaaccctccg
      841 agctggagct ggatgatgtc gtcatcacca acccccacat cgaggccatc ctggagaatg
      901 aggactggat cgaagatgcc tcgggcctca tgtcccactg catcgccatc ttgaagattt
      961 gtcacactct gacagaaaaa ctcgttgcca tgacaatggg ttctggggcc aagatgaaga
     1021 cgtcagcaag tgtcagtgac atcattgtgg tggccaaacg gattagcccc agagtggacg
     1081 acgtcgtgaa gtcaatgtac cctccactgg accccaagct cctggatgca cggacaaccg
     1141 ccctgctgct gtccgttagt cacttggtgc tagtgaccag gaacgcctgc catctaaccg
     1201 ggggcctgga ctggattgac caatcactgt ctgccgctga ggagcacctg gaagtccttc
     1261 gagaggcagc cctggcttct gagccagata aaagcctccc caaccctgag ggcttcctgc
     1321 aggaacagtc ggccattgca accatgggga tccaaccggt cgccaccatg gtgagcaagg
     1381 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1441 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1501 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1561 tgtcctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     1621 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1681 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1741 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1801 actactttag cgacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca
     1861 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc
     1921 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1981 agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2041 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgagcg gccgcgactc
     2101 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2161 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2221 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2281 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     2341 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2401 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2461 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2521 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2581 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2641 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2701 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2761 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2821 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2881 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2941 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3001 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3061 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3121 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3181 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3241 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3301 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3361 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3421 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3481 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3541 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3601 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3661 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3721 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3781 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3841 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3901 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3961 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4021 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4081 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4141 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4201 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4261 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4321 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4381 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4441 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4501 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4561 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4621 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4681 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4741 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4801 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4861 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4921 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4981 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5041 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5101 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5161 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5221 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5281 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5341 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5401 aaccgtatta ccgccatgca t
//
