LOCUS       sequence-254376-        5415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sGFP2-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 5415)
  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 5415)
  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-sGFP2
FEATURES             Location/Qualifiers
     source          1..5415
                     /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"
     CDS             join(1362..1364,1365..1367,1368..2078)
                     /codon_start=1
                     /product="strongly enhanced green fluorescent protein 
                     (Kremers et al., 2007)"
                     /label=SGFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVSILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQYDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGTTLGMDELYK"
     primer_bind     complement(1407..1428)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1668..1687)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2015..2036
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2203..2324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2240..2259)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2294..2313
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2331..2786)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2468..2489)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2680..2699
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2813..2917
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2919..3276
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2919..3110
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2942..2962)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3127..3262
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3311..4105
                     /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             3311..4105
                     /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(3365..3384)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3975..3994
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4293..4312)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4337..4384
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4713..5301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5202..5221
                     /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 catggtgagc aagggcgagg
     1381 agctgttcac cggggtggtg tccatcctgg tcgagctgga cggcgacgta aacggccaca
     1441 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1501 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1561 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagtacgac ttcttcaagt
     1621 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1681 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1741 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1801 acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1861 agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca
     1921 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca
     1981 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2041 ccgccgggac cactctcggc atggacgagc tgtacaagtg agcggccgcg actctagatc
     2101 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2161 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2221 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2281 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     2341 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2401 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2461 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2521 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2581 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2641 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2701 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2761 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     2821 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2881 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     2941 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3001 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3061 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3121 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3181 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3241 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     3301 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3361 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3421 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3481 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3541 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3601 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3661 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3721 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3781 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     3841 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3901 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3961 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4021 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4081 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4141 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4201 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4261 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     4321 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     4381 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4441 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4501 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     4561 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4621 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4681 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4741 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4801 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4861 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4921 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4981 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5041 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5101 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5161 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5221 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5281 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5341 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5401 attaccgcca tgcat
//
