LOCUS       sequence-190626-        5803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of mutant TMEM230 (isoform 1) and ZsGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5803)
  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 5803)
  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-M1/R141L-IRES(isoform 1)
FEATURES             Location/Qualifiers
     source          1..5803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     495..514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        769..1072
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1073..1276
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1227..1247
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1895..2469,2470..2472,2473..2481)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2062..2079)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2289..2308
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2482..3177
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    3299..3420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3336..3355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3390..3409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3427..3882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3564..3585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3776..3795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3909..4013
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4015..4372
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4015..4206
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4038..4058)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4223..4358
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4407..5201
                     /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             4407..5201
                     /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(4461..4480)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5071..5090
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5389..5408)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5433..5480
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
       61 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
      181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      661 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
      721 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      781 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      841 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      901 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      961 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     1021 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     1081 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     1141 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     1201 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     1261 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc ggactcagat
     1321 ctcgaggcca ccatgcaacc ttgggcgctg ccaaccgtgg gcgagctctg ggtgtgcggg
     1381 cggcctggcg cggcgctccg atggagtcta gttctgtcgc ccaggctgga gcccagtggt
     1441 gtgatctcag ctcactgcaa tctccacctc ctggcttcaa gcgattcttc tgcttcagcc
     1501 tcccgactgt gtcagcgtgt tatgatgccg tcccgtacca acctggctac tggaatcccc
     1561 agtagtaaag tgaaatattc aaggctctcc agcacagacg atggctacat tgaccttcag
     1621 tttaagaaaa cccctcctaa gatcccttat aaggccatcg cacttgccac tgtgctgttt
     1681 ttgattggcg cctttctcat tattataggc tccctcctgc tgtcaggcta catcagcaaa
     1741 gggggggcag acctggccgt tccagtgctg atcattggca ttctggtgtt cctacccgga
     1801 ttttaccacc tgcgcatcgc ttactatgca tccaaaggct accgtggtta ctcctatgat
     1861 gacattccag actttgatga ctagcacgga tccgcccctc tccctccccc ccccctaacg
     1921 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1981 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     2041 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     2101 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     2161 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     2221 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     2281 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     2341 cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga
     2401 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     2461 gatgataata tggccacaac catggcccag tccaagcacg gcctgaccaa ggagatgacc
     2521 atgaagtacc gcatggaggg ctgcgtggac ggccacaagt tcgtgatcac cggcgagggc
     2581 atcggctacc ccttcaaggg caagcaggcc atcaacctgt gcgtggtgga gggcggcccc
     2641 ttgcccttcg ccgaggacat cttgtccgcc gccttcatgt acggcaaccg cgtgttcacc
     2701 gagtaccccc aggacatcgt cgactacttc aagaactcct gccccgccgg ctacacctgg
     2761 gaccgctcct tcctgttcga ggacggcgcc gtgtgcatct gcaacgccga catcaccgtg
     2821 agcgtggagg agaactgcat gtaccacgag tccaagttct acggcgtgaa cttccccgcc
     2881 gacggccccg tgatgaagaa gatgaccgac aactgggagc cctcctgcga gaagatcatc
     2941 cccgtgccca agcagggcat cttgaagggc gacgtgagca tgtacctgct gctgaaggac
     3001 ggtggccgct tgcgctgcca gttcgacacc gtgtacaagg ccaagtccgt gccccgcaag
     3061 atgcccgact ggcacttcat ccagcacaag ctgacccgcg aggaccgcag cgacgccaag
     3121 aaccagaagt ggcacctgac cgagcacgcc atcgcctccg gctccgcctt gccctgagcg
     3181 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3241 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3301 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3361 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3421 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3481 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3541 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3601 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3661 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3721 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3781 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     3841 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     3901 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3961 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4021 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4081 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4201 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4261 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4321 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4381 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4441 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4501 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4561 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4621 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4681 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4741 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4801 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4861 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4921 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4981 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5041 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5101 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5161 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5221 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5281 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5341 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5401 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5461 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5521 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5581 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5641 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa gga
//
