LOCUS       Exported                6835 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 6835)
  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 6835)
  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..6835
                     /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"
     misc_feature    944..950
                     /label=KOZAK
     CDS             947..1543
                     /codon_start=1
                     /label=hPKD1(4106-4302)
                     /translation="MARWRYHALRGELYRPAWEPQDYEMVELFLRRLRLWMGLSKVKEF
                     RHKVRFEGMEPLPSRSSRGSKVSPDVPPPSAGSDASHPSTSSSQLDGLSVSLGRLGTRC
                     EPEPSRLQAVFEALLTQFDRLNQATEDVYQLEQQLHSLQGRRSSRAPAGSSRGPSPGLR
                     PALPSRLARASRGVDLATGPSRTPLRAKNKVHPSST"
     CDS             950..952
                     /codon_start=1
                     /product="additional Ala which is not part of PC1"
                     /label=Add Ala
                     /translation="A"
     misc_feature    1545..2387
                     /label=CL389-gBLOCK
     CDS             1556..2272
                     /codon_start=1
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2282..2308
                     /label=HA tag
     CDS             2312..2338
                     /label=HA tag
     CDS             2342..2368
                     /label=HA tag
     misc_feature    2373..2383
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(2387..2405)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2431..2655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2701..3129
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3143..3473
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3324..3459
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3540..4334
                     /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    4508..4629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4678..4694)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(5080..5668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5839..6699)
                     /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(6700..6804)
                     /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 ttcaccatgg cccgctggcg
      961 ctaccacgct ttgcgtggag agctgtaccg gccggcctgg gagccccagg actacgagat
     1021 ggtggagttg ttcctgcgca ggctgcgcct ctggatgggc ctcagcaagg tcaaggagtt
     1081 ccgccacaaa gtccgctttg aagggatgga gccgctgccc tctcgctcct ccaggggctc
     1141 caaggtatcc ccggatgtgc ccccacccag cgctggctcc gatgcctcgc acccctccac
     1201 ctcctccagc cagctggatg ggctgagcgt gagcctgggc cggctgggga caaggtgtga
     1261 gcccgagccc tcccgcctcc aagccgtgtt cgaggccctg ctcacccagt ttgaccgact
     1321 caaccaggcc acagaggacg tctaccagct ggagcagcag ctgcacagcc tgcaaggccg
     1381 caggagcagc cgggcgcccg ccggatcttc ccgtggccca tccccgggcc tgcggccagc
     1441 actgcccagc cgccttgccc gggccagtcg gggtgtggac ctggccactg gccccagcag
     1501 gacacccctt cgggccaaga acaaggtcca ccccagcagc actctcgaga tcgatatggt
     1561 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1621 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1681 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1741 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1801 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1861 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1921 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1981 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2041 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2101 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2161 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2221 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agatcgatat
     2281 gtacccatac gatgttccag attacgctgg ctatccctat gacgtcccgg actatgcagg
     2341 atatccatat gacgttccag attacgctta atctagaggg ccctattcta tagtgtcacc
     2401 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2461 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2521 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2581 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2641 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2701 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2761 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2821 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     2881 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2941 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3001 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3061 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3121 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3181 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3241 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3301 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3361 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3421 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3481 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     3541 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3601 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3661 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3721 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3781 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3841 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3901 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3961 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4021 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4081 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4141 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4201 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4261 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4321 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4381 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4441 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4501 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4561 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4621 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4681 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4741 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4801 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4861 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5521 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6121 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6421 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
