LOCUS Exported 5821 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION mammalian expression of mutant TMEM230 (isoform 1) and ZsGreen. ACCESSION . VERSION . KEYWORDS TMEM230-M2/X184W-IRES(isoform 1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5821) 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 5821) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5821 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 24..612 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 513..532 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 787..1090 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1091..1294 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1245..1265 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 1931..2517 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(2098..2115) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2325..2344 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2518..3213 /codon_start=1 /product="Zoanthus green fluorescent protein" /label=ZsGreen1 /note="mammalian codon-optimized" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" polyA_signal 3335..3456 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3372..3391) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3426..3445 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3463..3918) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3600..3621) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3812..3831 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3945..4049 /gene="bla" /label=AmpR promoter promoter 4051..4408 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4074..4094) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4259..4394 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4321..4340 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4443..5237 /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(4497..4516) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5107..5126 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5425..5444) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5469..5516 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 61 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg 181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 661 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 721 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 781 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 841 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 901 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 961 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 1021 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 1081 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 1141 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 1201 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 1261 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta 1321 gcgctaccgg actcagatct cgaggccacc atgcaacctt gggcgctgcc aaccgtgggc 1381 gagctctggg tgtgcgggcg gcctggcgcg gcgctccgat ggagtctagt tctgtcgccc 1441 aggctggagc ccagtggtgt gatctcagct cactgcaatc tccacctcct ggcttcaagc 1501 gattcttctg cttcagcctc ccgactgtgt cagcgtgtta tgatgccgtc ccgtaccaac 1561 ctggctactg gaatccccag tagtaaagtg aaatattcaa ggctctccag cacagacgat 1621 ggctacattg accttcagtt taagaaaacc cctcctaaga tcccttataa ggccatcgca 1681 cttgccactg tgctgttttt gattggcgcc tttctcatta ttataggctc cctcctgctg 1741 tcaggctaca tcagcaaagg gggggcagac cgggccgttc cagtgctgat cattggcatt 1801 ctggtgttcc tacccggatt ttaccacctg cgcatcgctt actatgcatc caaaggctac 1861 cgtggttact cctatgatga cattccagac tttgatgact ggcacccacc ccatagctga 1921 ggaggatccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat 1981 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg 2041 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc 2101 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt 2161 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg 2221 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac 2281 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg 2341 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg 2401 ggcctcggta cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc 2461 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg 2521 gcccagtcca agcacggcct gaccaaggag atgaccatga agtaccgcat ggagggctgc 2581 gtggacggcc acaagttcgt gatcaccggc gagggcatcg gctacccctt caagggcaag 2641 caggccatca acctgtgcgt ggtggagggc ggccccttgc ccttcgccga ggacatcttg 2701 tccgccgcct tcatgtacgg caaccgcgtg ttcaccgagt acccccagga catcgtcgac 2761 tacttcaaga actcctgccc cgccggctac acctgggacc gctccttcct gttcgaggac 2821 ggcgccgtgt gcatctgcaa cgccgacatc accgtgagcg tggaggagaa ctgcatgtac 2881 cacgagtcca agttctacgg cgtgaacttc cccgccgacg gccccgtgat gaagaagatg 2941 accgacaact gggagccctc ctgcgagaag atcatccccg tgcccaagca gggcatcttg 3001 aagggcgacg tgagcatgta cctgctgctg aaggacggtg gccgcttgcg ctgccagttc 3061 gacaccgtgt acaaggccaa gtccgtgccc cgcaagatgc ccgactggca cttcatccag 3121 cacaagctga cccgcgagga ccgcagcgac gccaagaacc agaagtggca cctgaccgag 3181 cacgccatcg cctccggctc cgccttgccc tgagcggccg cgactctaga tcataatcag 3241 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 3301 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 3361 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 3421 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata 3481 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg 3541 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc 3601 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa 3661 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt 3721 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac 3781 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta 3841 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg 3901 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 3961 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 4021 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt 4081 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 4141 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 4201 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 4261 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 4321 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg 4381 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc 4441 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 4501 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 4561 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 4621 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 4681 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 4741 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 4801 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 4861 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 4921 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg 4981 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 5041 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 5101 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 5161 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 5221 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 5281 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 5341 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 5401 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg 5461 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa 5521 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg 5581 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc 5641 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat 5701 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 5761 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 5821 c //