LOCUS       sequence-190624-        5804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of TMEM230 (isoform 1) and ZsGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5804)
  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 5804)
  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-WT-IRES(isoform 1)
FEATURES             Location/Qualifiers
     source          1..5804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      30..618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     519..538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        793..1096
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1097..1300
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1251..1271
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1919..2494,2495..2497,2498..2506)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2087..2104)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2314..2333
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2507..3202
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    3324..3445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3361..3380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3415..3434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3452..3907)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3589..3610)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3801..3820
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3934..4038
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4040..4397
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4040..4231
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4063..4083)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4248..4383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4432..5226
                     /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             4432..5226
                     /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(4486..4505)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5096..5115
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5414..5433)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5458..5505
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
       61 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
      181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      721 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      781 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      841 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      901 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      961 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1021 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1081 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     1141 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     1201 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     1261 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     1321 ccgctagcgc taccggactc agatctcgag gccaccatgc aaccttgggc gctgccaacc
     1381 gtgggcgagc tctgggtgtg cgggcggcct ggcgcggcgc tccgatggag tctagttctg
     1441 tcgcccaggc tggagcccag tggtgtgatc tcagctcact gcaatctcca cctcctggct
     1501 tcaagcgatt cttctgcttc agcctcccga ctgtgtcagc gtgttatgat gccgtcccgt
     1561 accaacctgg ctactggaat ccccagtagt aaagtgaaat attcaaggct ctccagcaca
     1621 gacgatggct acattgacct tcagtttaag aaaacccctc ctaagatccc ttataaggcc
     1681 atcgcacttg ccactgtgct gtttttgatt ggcgcctttc tcattattat aggctccctc
     1741 ctgctgtcag gctacatcag caaagggggg gcagaccggg ccgttccagt gctgatcatt
     1801 ggcattctgg tgttcctacc cggattttac cacctgcgca tcgcttacta tgcatccaaa
     1861 ggctaccgtg gttactccta tgatgacatt ccagactttg atgactagca cggatccgcc
     1921 cctctccctc cccccccccn taacgttact ggccgaagcc gcttggaata aggccggtgt
     1981 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     2041 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     2101 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     2161 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     2221 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     2281 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     2341 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtac
     2401 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     2461 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg cccagtccaa
     2521 gcacggcctg accaaggaga tgaccatgaa gtaccgcatg gagggctgcg tggacggcca
     2581 caagttcgtg atcaccggcg agggcatcgg ctaccccttc aagggcaagc aggccatcaa
     2641 cctgtgcgtg gtggagggcg gccccttgcc cttcgccgag gacatcttgt ccgccgcctt
     2701 catgtacggc aaccgcgtgt tcaccgagta cccccaggac atcgtcgact acttcaagaa
     2761 ctcctgcccc gccggctaca cctgggaccg ctccttcctg ttcgaggacg gcgccgtgtg
     2821 catctgcaac gccgacatca ccgtgagcgt ggaggagaac tgcatgtacc acgagtccaa
     2881 gttctacggc gtgaacttcc ccgccgacgg ccccgtgatg aagaagatga ccgacaactg
     2941 ggagccctcc tgcgagaaga tcatccccgt gcccaagcag ggcatcttga agggcgacgt
     3001 gagcatgtac ctgctgctga aggacggtgg ccgcttgcgc tgccagttcg acaccgtgta
     3061 caaggccaag tccgtgcccc gcaagatgcc cgactggcac ttcatccagc acaagctgac
     3121 ccgcgaggac cgcagcgacg ccaagaacca gaagtggcac ctgaccgagc acgccatcgc
     3181 ctccggctcc gccttgccct gagcggccgc gactctagat cataatcagc cataccacat
     3241 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3301 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3361 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3421 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3481 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3541 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3601 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3661 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3721 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3781 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3841 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3901 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3961 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4021 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4081 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4141 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4201 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4261 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4321 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4381 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4441 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4501 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4561 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4621 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4681 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4741 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4801 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4861 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4921 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     4981 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5041 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5101 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5161 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5221 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5281 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5341 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5401 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5461 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5521 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5581 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5641 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtca
//
