LOCUS       sequence-190784-        5614 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of 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-WT-IRES(isoform 2)
FEATURES             Location/Qualifiers
     source          1..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        777..1080
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1081..1284
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1235..1255
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1714..2288,2289..2291,2292..2300)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1881..1898)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2108..2127
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2301..2996
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    3118..3239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3155..3174)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3209..3228
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3246..3701)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3383..3404)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3595..3614
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3728..3832
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3834..4191
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3834..4025
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3857..3877)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4042..4177
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4226..5020
                     /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             4226..5020
                     /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(4280..4299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4890..4909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5208..5227)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5252..5299
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
      721 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      781 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      841 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      901 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      961 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1021 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1081 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     1141 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     1201 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     1261 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
     1321 actcagatct cgaggccacc atgatgccgt cccgtaccaa cctggctact ggaatcccca
     1381 gtagtaaagt gaaatattca aggctctcca gcacagacga tggctacatt gaccttcagt
     1441 ttaagaaaac ccctcctaag atcccttata aggccatcgc acttgccact gtgctgtttt
     1501 tgattggcgc ctttctcatt attataggct ccctcctgct gtcaggctac atcagcaaag
     1561 ggggggcaga ccgggccgtt ccagtgctga tcattggcat tctggtgttc ctacccggat
     1621 tttaccacct gcgcatcgct tactatgcat ccaaaggcta ccgtggttac tcctatgatg
     1681 acattccaga ctttgatgac tagcacggat ccgcccctct ccctcccccc cccctaacgt
     1741 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     1801 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     1861 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     1921 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     1981 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     2041 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     2101 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     2161 ccattgtatg ggatctgatc tggggcctcg gtacacatgc tttacatgtg tttagtcgag
     2221 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     2281 atgataatat ggccacaacc atggcccagt ccaagcacgg cctgaccaag gagatgacca
     2341 tgaagtaccg catggagggc tgcgtggacg gccacaagtt cgtgatcacc ggcgagggca
     2401 tcggctaccc cttcaagggc aagcaggcca tcaacctgtg cgtggtggag ggcggcccct
     2461 tgcccttcgc cgaggacatc ttgtccgccg ccttcatgta cggcaaccgc gtgttcaccg
     2521 agtaccccca ggacatcgtc gactacttca agaactcctg ccccgccggc tacacctggg
     2581 accgctcctt cctgttcgag gacggcgccg tgtgcatctg caacgccgac atcaccgtga
     2641 gcgtggagga gaactgcatg taccacgagt ccaagttcta cggcgtgaac ttccccgccg
     2701 acggccccgt gatgaagaag atgaccgaca actgggagcc ctcctgcgag aagatcatcc
     2761 ccgtgcccaa gcagggcatc ttgaagggcg acgtgagcat gtacctgctg ctgaaggacg
     2821 gtggccgctt gcgctgccag ttcgacaccg tgtacaaggc caagtccgtg ccccgcaaga
     2881 tgcccgactg gcacttcatc cagcacaagc tgacccgcga ggaccgcagc gacgccaaga
     2941 accagaagtg gcacctgacc gagcacgcca tcgcctccgg ctccgccttg ccctgagcgg
     3001 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     3061 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     3121 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3181 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3241 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3301 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3361 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3421 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3481 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3541 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3601 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3661 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3721 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3781 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3841 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3901 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4021 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4081 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4141 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4201 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4261 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4321 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4381 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4441 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4501 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4561 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4621 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4681 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4741 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4801 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4861 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4921 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4981 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5041 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     5101 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5161 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5221 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5281 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5341 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5401 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5461 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5521 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5581 tcgttccact gagcgtcaga ccccgtagaa aaga
//
