LOCUS       sequence-190794-        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-M1/R141L-IRES(isoform 2)
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        780..1083
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1084..1287
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1238..1258
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1717..2291,2292..2294,2295..2303)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1884..1901)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2111..2130
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2304..2999
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    3121..3242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3158..3177)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3212..3231
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3249..3704)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3386..3407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3598..3617
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3731..3835
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3837..4194
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3837..4028
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3860..3880)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4045..4180
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4229..5023
                     /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             4229..5023
                     /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(4283..4302)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4893..4912
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5211..5230)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5255..5302
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
      721 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      781 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      841 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      901 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      961 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     1021 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1081 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     1141 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     1201 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     1261 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
     1321 cggactcaga tctcgaggcc accatgatgc cgtcccgtac caacctggct actggaatcc
     1381 ccagtagtaa agtgaaatat tcaaggctct ccagcacaga cgatggctac attgaccttc
     1441 agtttaagaa aacccctcct aagatccctt ataaggccat cgcacttgcc actgtgctgt
     1501 ttttgattgg cgcctttctc attattatag gctccctcct gctgtcaggc tacatcagca
     1561 aagggggggc agacctggcc gttccagtgc tgatcattgg cattctggtg ttcctacccg
     1621 gattttacca cctgcgcatc gcttactatg catccaaagg ctaccgtggt tactcctatg
     1681 atgacattcc agactttgat gactagcacg gatccgcccc tctccctccc ccccccctaa
     1741 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     1801 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     1861 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     1921 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     1981 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     2041 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     2101 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     2161 accccattgt atgggatctg atctggggcc tcggtacaca tgctttacat gtgtttagtc
     2221 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     2281 acgatgataa tatggccaca accatggccc agtccaagca cggcctgacc aaggagatga
     2341 ccatgaagta ccgcatggag ggctgcgtgg acggccacaa gttcgtgatc accggcgagg
     2401 gcatcggcta ccccttcaag ggcaagcagg ccatcaacct gtgcgtggtg gagggcggcc
     2461 ccttgccctt cgccgaggac atcttgtccg ccgccttcat gtacggcaac cgcgtgttca
     2521 ccgagtaccc ccaggacatc gtcgactact tcaagaactc ctgccccgcc ggctacacct
     2581 gggaccgctc cttcctgttc gaggacggcg ccgtgtgcat ctgcaacgcc gacatcaccg
     2641 tgagcgtgga ggagaactgc atgtaccacg agtccaagtt ctacggcgtg aacttccccg
     2701 ccgacggccc cgtgatgaag aagatgaccg acaactggga gccctcctgc gagaagatca
     2761 tccccgtgcc caagcagggc atcttgaagg gcgacgtgag catgtacctg ctgctgaagg
     2821 acggtggccg cttgcgctgc cagttcgaca ccgtgtacaa ggccaagtcc gtgccccgca
     2881 agatgcccga ctggcacttc atccagcaca agctgacccg cgaggaccgc agcgacgcca
     2941 agaaccagaa gtggcacctg accgagcacg ccatcgcctc cggctccgcc ttgccctgag
     3001 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3061 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3121 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3181 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3241 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3301 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3361 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3421 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3481 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3541 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3601 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3661 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3721 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3781 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3841 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3901 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3961 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4021 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4081 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4141 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4201 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4261 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4321 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4381 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4441 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4501 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4561 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4621 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4681 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4741 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4801 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4861 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4921 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4981 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5041 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5101 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5161 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5221 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5281 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5341 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5401 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5461 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5521 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5581 ttttcgttcc actgagcgtc agaccccgta gaaa
//
