LOCUS       sequence-402598-        9416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for human AAVS1 locus knock-in, constitutive 
            expression of MitoTrap-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9416)
  AUTHORS   Fung HKH, Hayashi Y, Salo VT, Babenko A, Zagoriy I, Brunner A, 
            Ellenberg J, Muller CW, Cuylen-Haering S, Mahamid J
  TITLE     Genetically encoded multimeric tags for subcellular protein 
            localization in cryo-EM.
  JOURNAL   Nat Methods. 2023 Dec;20(12):1900-1908. doi: 
            10.1038/s41592-023-02053-0. Epub 2023 Nov 6.
  PUBMED    37932397
REFERENCE   2  (bases 1 to 9416)
  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/s41592-023-02053-0"; journalName: "Nat Methods"; date: 
            "2023-12"; volume: "20"; issue: "12"; pages: "1900-1908"
COMMENT     Sequence Label: pAAVS1_EF1a-MitoTrap-mCherry-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..9416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(226..330)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      357..812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(444..463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     654..675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    1329..2132
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     polyA_signal    2158..2206
                     /note="synthetic polyadenylation signal"
     misc_feature    2220..2311
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2260..2279
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2323..3483
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2552..3474
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3431..3451
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          3522..3654
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3637..3656
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        3699..3717
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3885..3894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3891..4595
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELY"
     primer_bind     complement(4036..4054)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4304..4323
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4599..4625
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4644..4922
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             4656..4925
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     misc_feature    4957..5523
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4973..5525
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5117..5134)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5344..5363
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5525..6127
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6024..6044
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6361..6442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6398..6417)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    6470..7306
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(7851..7868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8022..8610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8102..8121)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8781..9416,1..156,157..225))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMESDKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     join(9404..9416,1..7)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
       61 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
      121 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
      181 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
      241 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
      301 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
      361 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      421 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      481 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      541 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      601 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      661 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      721 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      781 gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc tgcgcaactg
      841 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc taccatgata
      901 agtaagtaat attaaggtac gggaggtact tggagcggcc gtcgacctac tctcttccgc
      961 attggagtcg ctttaactgg ccctggcttt ggcagcctgt gctgacccat gcagtcctcc
     1021 ttaccatccc tccctcgact tcccctcttc cgatgttgag cccctccagc cggtcctgga
     1081 ctttgtctcc ttccctgccc tgccctctcc tgaacctgag ccagctccca tagctcagtc
     1141 tggtctatct gcctggccct ggccattgtc actttgcgct gccctcctct cgcccccgag
     1201 tgcccttgct gtgccgccgg aactctgccc tctaacgctg ccgtctctct cctgagtccg
     1261 gaccactttg agctctactg gcttctgcgc cgcctctggc ccactgtttc cccttcccag
     1321 gcaggtcctg ctttctctga cctgcattct ctcccctggg cctgtgccgc tttctgtctg
     1381 cagcttgtgg cctgggtcac ctctacggct ggcccagatc cttccctgcc gcctccttca
     1441 ggttccgtct tcctccactc cctcttcccc ttgctctctg ctgtgttgct gcccaaggat
     1501 gctctttccg gagcacttcc ttctcggcgc tgcaccacgt gatgtcctct gagcggatcc
     1561 tccccgtgtc tgggtcctct ccgggcatct ctcctccctc acccaacccc atgccgtctt
     1621 cactcgctgg gttccctttt ccttctcctt ctggggcctg tgccatctct cgtttcttag
     1681 gatggccttc tccgacggat gtctcccttg cgtcccgcct ccccttcttg taggcctgca
     1741 tcatcaccgt ttttctggac aaccccaaag taccccgtct ccctggcttt agccacctct
     1801 ccatcctctt gctttctttg cctggacacc ccgttctcct gtggattcgg gtcacctctc
     1861 actcctttca tttgggcagc tcccctaccc cccttacctc tctagtctgt gctagctctt
     1921 ccagccccct gtcatggcat cttccagggg tccgagagct cagctagtct tcttcctcca
     1981 acccgggccc ctatgtccac ttcaggacag catgtttgct gcctccaggg atcctgtgtc
     2041 cccgagctgg gaccacctta tattcccagg gccggttaat gtggctctgg ttctgggtac
     2101 ttttatctgt cccctccacc ccacagtggg gcgcatgcgt ggaattatca cctcgagaat
     2161 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta
     2221 acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc
     2281 agtgcaagtg caggtgccag aacatttctc tactagtcgt gaggctccgg tgcccgtcag
     2341 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     2401 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     2461 cgcctttttc ccgagggtgg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     2521 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
     2581 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctccagta
     2641 cgtgattctt gatcccgagc tggagccagg ggcgggcctt gcgctttagg agccccttcg
     2701 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     2761 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     2821 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca ggatctgcac
     2881 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     2941 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     3001 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
     3061 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     3121 gctccagggg gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     3181 acacaaagga aaggggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     3241 cgggcgccgt ccaggcacct cgattagttc tggagctttt ggagtacgtc gtctttaggt
     3301 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     3361 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     3421 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     3481 tgaacacggg ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa
     3541 gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt
     3601 tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc
     3661 actcccagtt caattacagc tcttaaggct agagtactta atacgactca ctataggcta
     3721 gcgctaccgg actcagatct acgcgtttcg aagtatacga attcgccacc atgaagagct
     3781 tcattacaag gaacaagaca gccattttgg caaccgttgc tgctacaggt actgccatcg
     3841 gtgcctacta ttattacaac caattgcaac aggatccacc ggtcgccacc atggtgagca
     3901 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     3961 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     4021 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     4081 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     4141 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     4201 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     4261 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     4321 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     4381 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     4441 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     4501 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     4561 gccgccactc caccggcggc atggacgagc tgtacactta cccctacgac gtgcccgact
     4621 acgcaggctc cgcacgagtg gccatcctct ggcatgagat gtggcatgaa ggcctggaag
     4681 aggcatctcg tttgtacttt ggggaaagga acgtgaaagg catgtttgag gtgctggagc
     4741 ccttgcatgc tatgatggaa cggggccccc agactctgaa ggaaacatcc tttaatcagg
     4801 cctatggtcg agatttaatg gaggcccaag agtggtgcag gaagtacatg aaatcaggga
     4861 atgtcaagga cctcacccaa gcctgggacc tctattatca tgtgttccga cgaatctcaa
     4921 agcaggccgg ctgataagcg gccgctctag cccaattccg cccccccccc taacgttact
     4981 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
     5041 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     5101 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     5161 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     5221 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     5281 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     5341 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     5401 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     5461 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     5521 taatatgggc accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     5581 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     5641 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     5701 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     5761 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     5821 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     5881 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     5941 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     6001 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     6061 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     6121 tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgg
     6181 ctccgaccga agccacccgg ggcggccccg ccgaccccgc acccgccccc gaggcccacc
     6241 gactctagct agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
     6301 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     6361 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6421 aataaagcat ttttttcact gcattctagt tgtggtttgt ccatgcatca ctagggacag
     6481 gattggtgac agaaaagccc catccttagg cctcctcctt cctagtctcc tgatattggg
     6541 tctaaccccc acctcctgtt aggcagattc cttatctggt gacacacccc catttcctgg
     6601 agccatctct ctccttgcca gaacctctaa ggtttgctta cgatggagcc agagaggatc
     6661 ctgggaggga gagcttggca gggggtggga gggaaggggg ggatgcgtga cctgcccggt
     6721 tctcagtggc caccctgcgc taccctctcc cagaacctga gctgctctga cgcggctgtc
     6781 tggtgcgttt cactgatcct ggtgctgcag cttccttaca cttcccaaga ggagaagcag
     6841 tttggaaaaa caaaatcaga ataagttggt cctgagttct aactttggct cttcaccttt
     6901 ctagtcccca atttatattg ttcctccgtg cgtcagtttt acctgtgaga taaggccagt
     6961 agccagcccc gtcctggcag ggctgtggtg aggagggggg tgtccgtgtg gaaaactccc
     7021 tttgtgagaa tggtgcgtcc taggtgttca ccaggtcgtg gccgcctcta ctccctttct
     7081 ctttctccat ccttctttcc ttaaagagtc cccagtgcta tctgggacat attcctccgc
     7141 ccagagcagg gtcccgcttc cctaaggccc tgctctgggc ttctgggttt gagtccttgg
     7201 caagcccagg agaggcgctc aggcttccct gtcccccttc ctcgtccacc atctcatgcc
     7261 cctggctctc ctgccccttc cctacagggg ttcctggctc tgctcttcag actgagcccc
     7321 gttcccctgc atccccgttc ccctgcatcc cccttcccct gcatccccca gaggccccag
     7381 gccacctact tggcctggac cccacgagag gccaccccag ccctgtctac caggctgcct
     7441 tttgggtgga ttctcctcca actgtggggt gactgcttgg caaactcact cttcggggta
     7501 tcccaggagg cctggagcat tggggtgggc tggggttcag agaggaggga ttcccttctc
     7561 aggttacgtg gccaagaagc aggggagctg ggtttgggtc aggtctgggt gtggggtgac
     7621 cagcttatgc tgtttgccca ggacagccta gttttagcgc tgaaaccctc agtcctagga
     7681 aaacagggat ggttggtcac tgtctctggg ctcgaccgat gcccttgaga gccttcaacc
     7741 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     7801 tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact
     7861 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7921 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7981 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8041 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8101 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8161 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8221 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8281 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8341 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8401 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8461 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8521 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8581 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8641 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8701 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8761 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8821 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8881 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8941 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     9001 actttatccg actccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9061 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9121 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9181 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9241 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9301 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9361 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgcc
//
