LOCUS       sequence-190801-        5632 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 5632)
  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 5632)
  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-M2/X184W-IRES(isoform 2)
FEATURES             Location/Qualifiers
     source          1..5632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        781..1084
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1085..1288
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1239..1259
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(1736..2310,2311..2313,2314..2322)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1903..1920)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2130..2149
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2323..3018
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     polyA_signal    3140..3261
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3177..3196)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3231..3250
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3268..3723)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3405..3426)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3617..3636
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3750..3854
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3856..4213
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3856..4047
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3879..3899)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4064..4199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4248..5042
                     /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             4248..5042
                     /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(4302..4321)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4912..4931
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5230..5249)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5274..5321
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
      721 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      781 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      841 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      901 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      961 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     1021 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     1081 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     1141 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     1201 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     1261 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
     1321 ccggactcag atctcgaggc caccatgatg ccgtcccgta ccaacctggc tactggaatc
     1381 cccagtagta aagtgaaata ttcaaggctc tccagcacag acgatggcta cattgacctt
     1441 cagtttaaga aaacccctcc taagatccct tataaggcca tcgcacttgc cactgtgctg
     1501 tttttgattg gcgcctttct cattattata ggctccctcc tgctgtcagg ctacatcagc
     1561 aaaggggggg cagaccgggc cgttccagtg ctgatcattg gcattctggt gttcctaccc
     1621 ggattttacc acctgcgcat cgcttactat gcatccaaag gctaccgtgg ttactcctat
     1681 gatgacattc cagactttga tgactggcac ccaccccata gctgaggagg atccgcccct
     1741 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     1801 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     1861 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     1921 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     1981 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2041 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     2101 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     2161 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat
     2221 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     2281 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggccca gtccaagcac
     2341 ggcctgacca aggagatgac catgaagtac cgcatggagg gctgcgtgga cggccacaag
     2401 ttcgtgatca ccggcgaggg catcggctac cccttcaagg gcaagcaggc catcaacctg
     2461 tgcgtggtgg agggcggccc cttgcccttc gccgaggaca tcttgtccgc cgccttcatg
     2521 tacggcaacc gcgtgttcac cgagtacccc caggacatcg tcgactactt caagaactcc
     2581 tgccccgccg gctacacctg ggaccgctcc ttcctgttcg aggacggcgc cgtgtgcatc
     2641 tgcaacgccg acatcaccgt gagcgtggag gagaactgca tgtaccacga gtccaagttc
     2701 tacggcgtga acttccccgc cgacggcccc gtgatgaaga agatgaccga caactgggag
     2761 ccctcctgcg agaagatcat ccccgtgccc aagcagggca tcttgaaggg cgacgtgagc
     2821 atgtacctgc tgctgaagga cggtggccgc ttgcgctgcc agttcgacac cgtgtacaag
     2881 gccaagtccg tgccccgcaa gatgcccgac tggcacttca tccagcacaa gctgacccgc
     2941 gaggaccgca gcgacgccaa gaaccagaag tggcacctga ccgagcacgc catcgcctcc
     3001 ggctccgcct tgccctgagc ggccgcgact ctagatcata atcagccata ccacatttgt
     3061 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3121 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3181 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3241 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3301 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3361 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3421 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3481 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     3541 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     3601 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     3661 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     3721 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3781 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3841 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     3901 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3961 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4021 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4081 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4141 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4201 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4261 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4321 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4381 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4441 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4501 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4561 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4621 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4681 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4741 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     4801 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4861 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4921 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4981 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5041 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5101 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5161 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5221 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5281 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5341 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5401 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5461 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     5521 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5581 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aa
//
