LOCUS       Exported                6376 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses human TMEM206.
ACCESSION   .
VERSION     .
KEYWORDS    pIRES2-hPAC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6376)
  AUTHORS   Yang J, Chen J, Del Carmen Vitery M, Osei-Owusu J, Chu J, Yu H, Sun 
            S, Qiu Z
  TITLE     PAC, an evolutionarily conserved membrane protein, is a 
            proton-activated chloride channel.
  JOURNAL   Science. 2019 Apr 26;364(6438):395-399. doi: 
            10.1126/science.aav9739. Epub 2019 Apr 25.
  PUBMED    31023925
REFERENCE   2  (bases 1 to 6376)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6376
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    1735..2321
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1902..1919)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2129..2148
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2322..3041
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2322..3038
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2367..2388)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2628..2647)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2975..2996
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3164..3285
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3201..3220)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3255..3274
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3292..3747)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3429..3450)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3641..3660
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3774..3878
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3880..4237
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3903..3923)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4088..4223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4150..4169
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4272..5066
                     /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(4326..4345)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4936..4955
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5254..5273)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5298..5345
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5674..6262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6163..6182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc atgatccggc aggagcgctc cacatcctac caggagctga
      661 gtgaggagtt ggtccaggtg gttgagaact cagagctggc agacgagcag gacaaggaga
      721 cggtcagagt ccaaggtccg ggtatcttac caggcctgga cagcgagtcc gcctccagca
      781 gcatccgctt cagcaaggcc tgcctgaaga acgtcttctc ggtcctactc atcttcatct
      841 acctgctgct catggctgtg gccgtcttcc tggtctaccg gaccatcaca gactttcgtg
      901 agaaactcaa gcaccctgtc atgtctgtgt cttacaagga agtggatcgc tatgatgccc
      961 caggtattgc cttgtacccc ggtcaggccc agttgctcag ctgtaagcac cattacgagg
     1021 tcattcctcc tctgacaagc cctggccagc cgggtgacat gaattgcacc acccagagga
     1081 tcaactacac ggaccccttc tccaatcaga ctgtgaaatc tgccctgatt gtccaggggc
     1141 cccgggaagt gaaaaagcgg gagctggtct tcctccagtt ccgcctgaac aagagtagtg
     1201 aggacttcag cgccattgat tacctcctct tctcttcttt ccaggagttc ctgcaaagcc
     1261 caaacagggt aggcttcatg caggcctgtg agagtgccta ttccagctgg aagttctctg
     1321 ggggcttccg cacctgggtc aagatgtcac tggtaaagac caaggaggag gatgggcggg
     1381 aagcagtgga gttccggcag gagacaagtg tggttaacta cattgaccag aggccagctg
     1441 ccaaaaaaag tgctcaattg ttttttgtgg tctttgaatg gaaagatcct ttcatccaga
     1501 aagtccaaga tatagtcact gccaatcctt ggaacacaat tgctcttctc tgtggcgcct
     1561 tcttggcatt atttaaagca gcagagtttg ccaaactgag tataaaatgg atgatcaaaa
     1621 ttagaaagag ataccttaaa agaagaggtc aggcaacgag ccacataagc tgagaattct
     1681 gcagtcgaca ccggtgcgat cgcacgcgtc cgcggcccgg gcgcgccgga tccgcccctc
     1741 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     1801 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     1861 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     1921 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     1981 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     2041 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     2101 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     2161 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtacacatg
     2221 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     2281 ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc aagggcgagg
     2341 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2401 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2461 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2521 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2581 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2641 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2701 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2761 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2821 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2881 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     2941 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     3001 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc gactctagat
     3061 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3121 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3181 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3241 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     3301 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3361 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3421 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3481 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3541 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3601 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3661 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3721 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3781 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3841 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3901 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3961 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4021 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4081 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4141 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4201 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4261 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4321 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4381 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4441 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4501 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4561 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4621 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4681 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4741 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4801 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4861 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4921 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4981 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5041 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5101 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5161 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5221 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5281 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5341 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5401 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5461 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5521 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5581 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5641 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5701 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5761 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5821 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5881 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5941 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6001 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6061 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6121 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6241 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6301 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6361 tattaccgcc atgcat
//
