LOCUS       Exported                7717 bp ds-DNA     circular SYN 14-FEB-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7717)
  AUTHORS   Lin CC, Kurashige M, Liu Y, Terabayashi T, Ishimoto Y, Wang T, 
            Choudhary V, Hobbs R, Liu LK, Lee PH, Outeda P, Zhou F, Restifo NP, 
            Watnick T, Kawano H, Horie S, Prinz W, Xu H, Menezes LF, Germino GG.
  TITLE     A cleavage product of Polycystin-1 is a mitochondrial matrix protein
            that affects mitochondria morphology and function when 
            heterologously expressed.
  JOURNAL   Sci Rep 2018;8:2743.
  PUBMED    29426897
REFERENCE   2  (bases 1 to 7717)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Feb 28, 2018 from SnapGene Viewer 4.1.6
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7717
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    889..942
                     /label=MCS
                     /note="multiple cloning site"
     CDS             947..2425
                     /codon_start=1
                     /label=CD16-CD7-hPKD1(4077-4302)
                     /translation="MWQLLLPTALLLLVSAGMRTEDLPKAVVFLEPQWYSVLEKDSVTL
                     KCQGAYSPEDNSTQWFHNESLISSQASSYFIDAATVNDSGEYRCQTNLSTLSDPVQLEV
                     HIGWLLLQAPRWVFKEEDPIHLRCHSWKNTALHKVTYLQNGKDRKYFHHNSDFHIPKAT
                     LKDSGSYFCRGLVGSKNVSSETVNITITQGLAVSTISSFSPPGADPPRASALPAPPTGS
                     ALPDPQTASALPDPPAASALPAALAVISFLLGLGLGVACVLARTRESWHLSPLLCVGLW
                     ALRLWGALRLGAVILRWRYHALRGELYRPAWEPQDYEMVELFLRRLRLWMGLSKVKEFR
                     HKVRFEGMEPLPSRSSRGSKVSPDVPPPSAGSDASHPSTSSSQLDGLSVSLGRLGTRCE
                     PEPSRLQAVFEALLTQFDRLNQATEDVYQLEQQLHSLQGRRSSRAPAGSSRGPSPGLRP
                     ALPSRLARASRGVDLATGPSRTPLRAKNKVHPSST"
     CDS             953..1563
                     /codon_start=1
                     /label=CD16
                     /translation="QLLLPTALLLLVSAGMRTEDLPKAVVFLEPQWYSVLEKDSVTLKC
                     QGAYSPEDNSTQWFHNESLISSQASSYFIDAATVNDSGEYRCQTNLSTLSDPVQLEVHI
                     GWLLLQAPRWVFKEEDPIHLRCHSWKNTALHKVTYLQNGKDRKYFHHNSDFHIPKATLK
                     DSGSYFCRGLVGSKNVSSETVNITITQGLAVSTISSFSPP"
     CDS             1571..1744
                     /codon_start=1
                     /label=CD7
                     /translation="PPRASALPAPPTGSALPDPQTASALPDPPAASALPAALAVISFLL
                     GLGLGVACVLART"
     CDS             1748..1960
                     /codon_start=1
                     /label=PKD1_Exon45
                     /translation="ESWHLSPLLCVGLWALRLWGALRLGAVILRWRYHALRGELYRPAW
                     EPQDYEMVELFLRRLRLWMGLSKVKE"
     CDS             1961..2425
                     /codon_start=1
                     /label=PKD1_Exon46
                     /translation="FRHKVRFEGMEPLPSRSSRGSKVSPDVPPPSAGSDASHPSTSSSQ
                     LDGLSVSLGRLGTRCEPEPSRLQAVFEALLTQFDRLNQATEDVYQLEQQLHSLQGRRSS
                     RAPAGSSRGPSPGLRPALPSRLARASRGVDLATGPSRTPLRAKNKVHPSST"
     CDS             2438..3154
                     /codon_start=1
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3164..3190
                     /label=HA tag
     CDS             3194..3220
                     /label=HA tag
     CDS             3224..3250
                     /label=HA tag
     misc_feature    3255..3265
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(3269..3287)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3313..3537
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3583..4011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4025..4355
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4206..4341
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4422..5216
                     /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)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5390..5511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5560..5576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(5962..6550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6721..7581)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7582..7686)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcaccatgt ggcagctgct
      961 cctcccaact gctctgctac ttctagtttc agctggcatg cggactgaag atctcccaaa
     1021 ggctgtggtg ttcctggagc ctcaatggta cagcgtgctt gagaaggaca gtgtgactct
     1081 gaagtgccag ggagcctact cccctgagga caattccaca cagtggtttc acaatgagag
     1141 cctcatctca agccaggcct cgagctactt cattgacgct gccacagtca acgacagtgg
     1201 agagtacagg tgccagacaa acctctccac cctcagtgac ccggtgcagc tagaagtcca
     1261 tatcggctgg ctgttgctcc aggcccctcg gtgggtgttc aaggaggaag accctattca
     1321 cctgaggtgt cacagctgga agaacactgc tctgcataag gtcacatatt tacagaatgg
     1381 caaagacagg aagtattttc atcataattc tgacttccac attccaaaag ccacactcaa
     1441 agatagcggc tcctacttct gcagggggct tgttgggagt aaaaatgtgt cttcagagac
     1501 tgtgaacatc accatcactc aaggtttggc agtgtcaacc atctcatcat tctctccacc
     1561 tggcgcggat ccaccaaggg cctctgccct ccctgcccca ccgacaggct ccgccctccc
     1621 tgacccgcag acagcctctg ccctccctga cccgccagca gcctctgccc tccctgcggc
     1681 cctggcggtg atctccttcc tcctcgggct gggcctgggg gtggcgtgtg tgctggcgag
     1741 gacgcgtgag tcctggcact tgtcacccct gctgtgtgtg gggctctggg cactgcggct
     1801 gtggggcgcc ctacggctgg gggctgttat tctccgctgg cgctaccacg ccttgcgtgg
     1861 agagctgtac cggccggcct gggagcccca ggactacgag atggtggagt tgttcctgcg
     1921 caggctgcgc ctctggatgg gcctcagcaa ggtcaaggag ttccgccaca aagtccgctt
     1981 tgaagggatg gagccgctgc cctctcgctc ctccaggggc tccaaggtat ccccggatgt
     2041 gcccccaccc agcgctggct ccgatgcctc gcacccctcc acctcctcca gccagctgga
     2101 tgggctgagc gtgagcctgg gccggctggg gacaaggtgt gagcctgagc cctcccgcct
     2161 ccaagccgtg ttcgaggccc tgctcaccca gtttgaccga ctcaaccagg ccacagagga
     2221 cgtctaccag ctggagcagc agctgcacag cctgcaaggc cgcaggagca gccgggcgcc
     2281 cgccggatct tcccgtggcc catccccggg cctgcggcca gcactgccca gccgccttgc
     2341 ccgggccagt cggggtgtgg acctggccac tggccccagc aggacacccc ttcgggccaa
     2401 gaacaaggtc caccccagca gcactctcga gatcgatatg gtgagcaagg gcgaggagct
     2461 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2521 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2581 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2641 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2701 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2761 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2821 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2881 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2941 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     3001 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     3061 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3121 cgggatcact ctcggcatgg acgagctgta caagatcgat atgtacccat acgatgttcc
     3181 agattacgct ggctatccct atgacgtccc ggactatgca ggatatccat atgacgttcc
     3241 agattacgct taatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     3301 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3361 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3421 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3481 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3541 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3601 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3661 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3721 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     3781 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3841 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3901 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
     3961 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4021 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
     4081 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4141 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4201 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4261 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4321 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4381 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4441 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4501 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4561 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4621 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4681 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4741 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4801 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4861 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4921 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4981 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     5041 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5101 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5161 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5221 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5281 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5341 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5401 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5461 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5521 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5581 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5641 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5701 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5761 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5821 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5881 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5941 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6001 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6061 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6121 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6181 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6241 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6301 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6361 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6421 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6481 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6541 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6601 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6661 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6721 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6781 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6841 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6901 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6961 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7021 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7081 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7141 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7201 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7261 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7321 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7381 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7441 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7501 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7561 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7621 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7681 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
