LOCUS       sequence-190806-        5614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of mutant TMEM230 (isoform 2) and ZsGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5614)
  AUTHORS   Deng HX, Shi Y, Yang Y, Ahmeti KB, Miller N, Huang C, Cheng L, Zhai 
            H, Deng S, Nuytemans K, Corbett NJ, Kim MJ, Deng H, Tang B, Yang Z, 
            Xu Y, Chan P, Huang B, Gao XP, Song Z, Liu Z, Fecto F, Siddique N, 
            Foroud T, Jankovic J, Ghetti B, Nicholson DA, Krainc D, Melen O, 
            Vance JM, Pericak-Vance MA, Ma YC, Rajput AH, Siddique T
  TITLE     Identification of TMEM230 mutations in familial Parkinson's disease.
  JOURNAL   Nat Genet. 2016 Jul;48(7):733-9. doi: 10.1038/ng.3589. Epub 2016 Jun
            6.
  PUBMED    27270108
REFERENCE   2  (bases 1 to 5614)
  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/ng.3589"; 
            journalName: "Nat Genet"; date: "2016-07"; volume: "48"; issue: "7";
            pages: "733-9"
COMMENT     Sequence Label: TMEM230-M3/Y92C-IRES(isoform 2)
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      22..610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     511..530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        785..1088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1089..1292
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1243..1263
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1722..2296,2297..2299,2300..2308)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1889..1906)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2116..2135
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2309..3004
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    3126..3247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3163..3182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3217..3236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3254..3709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3391..3412)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3603..3622
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3736..3840
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3842..4199
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3842..4033
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3865..3885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4050..4185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4234..5028
                     /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             4234..5028
                     /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(4288..4307)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4898..4917
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5216..5235)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5260..5307
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
       61 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
      181 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
      241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      661 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
      721 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      781 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      841 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      901 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      961 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     1021 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     1081 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     1141 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     1201 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     1261 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
     1321 gctaccggac tcagatctcg aggccaccat gatgccgtcc cgtaccaacc tggctactgg
     1381 aatccccagt agtaaagtga aatattcaag gctctccagc acagacgatg gctgcattga
     1441 ccttcagttt aagaaaaccc ctcctaagat cccttataag gccatcgcac ttgccactgt
     1501 gctgtttttg attggcgcct ttctcattat tataggctcc ctcctgctgt caggctacat
     1561 cagcaaaggg ggggcagacc gggccgttcc agtgctgatc attggcattc tggtgttcct
     1621 acccggattt taccacctgc gcatcgctta ctatgcatcc aaaggctacc gtggttactc
     1681 ctatgatgac attccagact ttgatgacta gcacggatcc gcccctctcc ctcccccccc
     1741 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     1801 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     1861 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     1921 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
     1981 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     2041 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     2101 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     2161 aaggtacccc attgtatggg atctgatctg gggcctcggt acacatgctt tacatgtgtt
     2221 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     2281 aaaacacgat gataatatgg ccacaaccat ggcccagtcc aagcacggcc tgaccaagga
     2341 gatgaccatg aagtaccgca tggagggctg cgtggacggc cacaagttcg tgatcaccgg
     2401 cgagggcatc ggctacccct tcaagggcaa gcaggccatc aacctgtgcg tggtggaggg
     2461 cggccccttg cccttcgccg aggacatctt gtccgccgcc ttcatgtacg gcaaccgcgt
     2521 gttcaccgag tacccccagg acatcgtcga ctacttcaag aactcctgcc ccgccggcta
     2581 cacctgggac cgctccttcc tgttcgagga cggcgccgtg tgcatctgca acgccgacat
     2641 caccgtgagc gtggaggaga actgcatgta ccacgagtcc aagttctacg gcgtgaactt
     2701 ccccgccgac ggccccgtga tgaagaagat gaccgacaac tgggagccct cctgcgagaa
     2761 gatcatcccc gtgcccaagc agggcatctt gaagggcgac gtgagcatgt acctgctgct
     2821 gaaggacggt ggccgcttgc gctgccagtt cgacaccgtg tacaaggcca agtccgtgcc
     2881 ccgcaagatg cccgactggc acttcatcca gcacaagctg acccgcgagg accgcagcga
     2941 cgccaagaac cagaagtggc acctgaccga gcacgccatc gcctccggct ccgccttgcc
     3001 ctgagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     3061 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3121 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3181 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3241 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3301 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3361 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3421 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3481 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3541 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3601 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3661 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3721 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3781 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3841 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3901 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3961 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4021 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4081 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4141 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4201 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4261 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4321 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4381 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4441 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4501 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4561 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4621 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4681 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4741 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4801 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4861 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4921 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4981 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5041 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5101 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5161 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5221 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5281 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5341 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5401 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5461 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5521 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5581 gtgagttttc gttccactga gcgtcagacc ccgt
//
