pMK395 (AAVS1 CMV-OsTIR1(F74A))

LOCUS       pMK395 (AAVS1 CMV-OsTIR1(F74A))  10098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of OsTIR1(F74A) in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10098)
  AUTHORS   Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano 
            R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, 
            Hayashi KI, Kanemaki MT
  TITLE     The auxin-inducible degron 2 technology provides sharp degradation 
            control in yeast, mammalian cells, and mice.
  JOURNAL   Nat Commun. 2020 Nov 11;11(1):5701. doi: 10.1038/s41467-020-19532-z.
  PUBMED    33177522
REFERENCE   2  (bases 1 to 10098)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2020-11-11"; volume: "11(1):5701. doi"; pages: " 
            10.1038/s41467-020-19532-z"
COMMENT     Sequence Label: pMK395 (AAVS1 CMV-OsTIR1(F74A))
FEATURES             Location/Qualifiers
     source          1..10098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    129..150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(165..170,171..188,189..195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    203..219
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     208..230
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     227..243
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     227..243
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        264..282
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    317..1120
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     enhancer        1175..1478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1479..1682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1633..1653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    3621..3742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3658..3677)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3712..3731
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    3769..3802
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3863..4362
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3898..3921)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4295..4314
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4402..5001
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4402..4421)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    5558..5591
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5622..6458
                     /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(6492..6511)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6493..6511)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6518..6535)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6518..6534)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6527..6549)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             6672..6974
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     6853..6872
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             7323..8117
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             7323..8117
                     /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(7377..7396)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7987..8006
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(8367..9158,9159..9227))
                     /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"
     primer_bind     8990..9009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9351..9939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9840..9859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(10093..10098,1..12)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
       61 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
      121 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
      181 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
      241 gaccatgatt acgccaagct cagaattaac cctcactaaa gggactagtc ctgcaggttt
      301 aaacgaattc gccctttgct ttctctgacc agcattctct cccctgggcc tgtgccgctt
      361 tctgtctgca gcttgtggcc tgggtcacct ctacggctgg cccagatcct tccctgccgc
      421 ctccttcagg ttccgtcttc ctccactccc tcttcccctt gctctctgct gtgttgctgc
      481 ccaaggatgc tctttccgga gcacttcctt ctcggcgctg caccacgtga tgtcctctga
      541 gcggatcctc cccgtgtctg ggtcctctcc gggcatctct cctccctcac ccaaccccat
      601 gccgtcttca ctcgctgggt tcccttttcc ttctccttct ggggcctgtg ccatctctcg
      661 tttcttagga tggccttctc cgacggatgt ctcccttgcg tcccgcctcc ccttcttgta
      721 ggcctgcatc atcaccgttt ttctggacaa ccccaaagta ccccgtctcc ctggctttag
      781 ccacctctcc atcctcttgc tttctttgcc tggacacccc gttctcctgt ggattcgggt
      841 cacctctcac tcctttcatt tgggcagctc ccctaccccc cttacctctc tagtctgtgc
      901 tagctcttcc agccccctgt catggcatct tccaggggtc cgagagctca gctagtcttc
      961 ttcctccaac ccgggcccct atgtccactt caggacagca tgtttgctgc ctccagggat
     1021 cctgtgtccc cgagctggga ccaccttata ttcccagggc cggttaatgt ggctctggtt
     1081 ctgggtactt ttatctgtcc cctccacccc acagtggggc aagctagtaa tcaattacgg
     1141 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     1201 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     1261 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     1321 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     1381 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     1441 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     1501 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     1561 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     1621 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     1681 ctggtttagt gaaccgtcag atccgctagc gctgccacca tgacatactt tcctgaagag
     1741 gtcgtcgaac acatttttag cttcctgcct gcacagagag atagaaacac agtgagcctg
     1801 gtctgcaaag tgtggtacga gatcgaacgc ctgagccgga gaggagtgtt cgtcggcaac
     1861 tgctatgctg tgagagcagg cagggtcgcc gctaggtttc caaatgtgcg cgcactgacc
     1921 gtcaagggga aaccccacgc cgccgacttt aacctggtgc cccctgattg gggaggatac
     1981 gccggccctt ggatcgaggc agccgctcgc ggctgtcatg gactggagga actgcgcatg
     2041 aagcgaatgg tggtctctga cgaaagtctg gagctgctgg ctcggagctt ccctaggttt
     2101 cgcgcactgg tgctgatttc ttgcgaaggc ttcagcaccg atggactggc agccgtggcc
     2161 tcccactgta agctgctgcg ggagctggac ctccaggaga atgaagtgga ggatagaggc
     2221 cccagatggc tgtcttgctt cccagactca tgtaccagcc tggtgtccct gaactttgcc
     2281 tgcatcaaag gcgaagtgaa tgctgggtcc ctggagcggc tggtctcaag aagccccaac
     2341 ctgaggtctc tgcggctgaa ccggagcgtg agcgtggaca ctctggctaa gattctgctg
     2401 agaaccccta acctggagga tctgggaacc ggcaatctga cagacgattt ccagacagaa
     2461 tcctacttta aactgacttc tgccctggag aagtgtaaaa tgctgaggag tctgtcagga
     2521 ttctgggatg cttcacccgt gtgcctgagc tttatctacc ctctgtgtgc acagctgaca
     2581 ggcctgaacc tgagctatgc accaaccctg gacgccagtg atctgacaaa gatgatctca
     2641 cgctgcgtga aactccagcg actgtgggtg ctggactgta tttccgataa ggggctccag
     2701 gtggtcgcca gctcctgcaa ggacctccag gagctgagag tgttcccatc tgatttttac
     2761 gtggccggat atagtgctgt cactgaggaa ggcctggtgg cagtctcact gggatgccca
     2821 aagctgaaca gcctgctgta tttctgtcat cagatgacta atgctgcact ggtgaccgtc
     2881 gccaagaact gccctaattt cacccgattt cggctgtgta ttctggaacc aggcaaaccc
     2941 gacgtggtca catcccagcc actggatgaa gggtttggag ctatcgtgag agagtgcaag
     3001 ggactccaga ggctgagcat ttccggcctg ctgacagaca aagtgttcat gtacatcggc
     3061 aagtatgcta agcagctgga gatgctgagc attgcatttg ccggagactc cgataagggc
     3121 atgatgcacg tgatgaacgg gtgtaagaat ctgcgaaaac tggaaatccg ggacagccct
     3181 ttcggggatg ccgctctgct gggaaacttt gccagatacg agacaatgag gagcctgtgg
     3241 atgtctagtt gcaatgtgac tctgaagggc tgtcaggtcc tggctagtaa aatgcctatg
     3301 ctgaacgtgg aagtcattaa tgagcgggac gggtctaacg aaatggagga aaatcatggc
     3361 gacctgccaa aggtggagaa actgtatgtg tatcggacca ccgcaggggc aagagatgat
     3421 gctcccaact ttgtgaagat tctgtgagga tccatcggat cccgggcccg tcgacggtac
     3481 cgcgggcccg ggatccaccg gggccgcgac tctagatcat aatcagccat accacatttg
     3541 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3601 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3661 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3721 ccaaactcat caatgtatct taagcgatca tattcaataa cccttaatat aacttcgtat
     3781 aatgtatgct atacgaagtt attaggtctg aagaggagtt tacgtccagc caagcttagg
     3841 atctcgacct cgaaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
     3901 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
     3961 cattccacat ccaccggtag gcgccaaccg actccgttct ttggtggccc cttcgcgcca
     4021 ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg
     4081 acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag
     4141 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc
     4201 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc
     4261 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     4321 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcat ccatctagat
     4381 ctcgagcagc tgaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     4441 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4501 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4561 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     4621 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4681 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4741 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4801 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     4861 gccggggtgc ccgccttcct ggagacctcc gcgccccaca acctcccctt ctacgagcgg
     4921 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4981 acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc
     5041 gcacgacccc atgcatcgat gatatcagat ccccgggatg cagaaattga tgatctatta
     5101 aacaataaag atgtccacta aaatggaagt ttttcctgtc atactttgtt aagaagggtg
     5161 agaacagagt acctacattt tgaatggaag gattggagct acgggggtgg gggtggggtg
     5221 ggattagata aatgcctgct ctttactgaa ggctctttac tattgcttta tgataatgtt
     5281 tcatagttgg atatcataat ttaaacaagc aaaaccaaat taagggccag ctcattcctc
     5341 ccactcatga tctatagatc tatagatctc tcgtgggatc attgtttttc tcttgattcc
     5401 cactttgtgg ttctaagtac tgtggtttcc aaatgtgtca gtttcatagc ctgaagaacg
     5461 agatcagcag cctctgttcc acatacactt cattctcagt attgttttgc caagttctaa
     5521 ttccatcaga agctggtcga gatccggaac ccttaatata acttcgtata atgtatgcta
     5581 tacgaagtta ttaggtccct cgaagaggtt cactaagctt tactagggac aggattggtg
     5641 acagaaaagc cccatcctta ggcctcctcc ttcctagtct cctgatattg ggtctaaccc
     5701 ccacctcctg ttaggcagat tccttatctg gtgacacacc cccatttcct ggagccatct
     5761 ctctccttgc cagaacctct aaggtttgct tacgatggag ccagagagga tcctgggagg
     5821 gagagcttgg cagggggtgg gagggaaggg ggggatgcgt gacctgcccg gttctcagtg
     5881 gccaccctgc gctaccctct cccagaacct gagctgctct gacgcggctg tctggtgcgt
     5941 ttcactgatc ctggtgctgc agcttcctta cacttcccaa gaggagaagc agtttggaaa
     6001 aacaaaatca gaataagttg gtcctgagtt ctaactttgg ctcttcacct ttctagtccc
     6061 caatttatat tgttcctccg tgcgtcagtt ttacctgtga gataaggcca gtagccagcc
     6121 ccgtcctggc agggctgtgg tgaggagggg ggtgtccgtg tggaaaactc cctttgtgag
     6181 aatggtgcgt cctaggtgtt caccaggtcg tggccgcctc tactcccttt ctctttctcc
     6241 atccttcttt ccttaaagag tccccagtgc tatctgggac atattcctcc gcccagagca
     6301 gggtcccgct tccctaaggc cctgctctgg gcttctgggt ttgagtcctt ggcaagccca
     6361 ggagaggcgc tcaggcttcc ctgtccccct tcctcgtcca ccatctcatg cccctggctc
     6421 tcctgcccct tccctacagg ggttcctggc tctgctctaa gggcgaattc gcggccgcta
     6481 aattcaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     6541 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6601 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6661 tatacgtacg gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
     6721 tggatgtaca gagtgatatt attgacacgc cggggcgacg gatggtgatc cccctggcca
     6781 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
     6841 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg
     6901 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
     6961 tctggggaat ataaatgtca ggcatgagat tatcaaaaag gatcttcacc tagatccttt
     7021 tcacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     7081 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
     7141 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     7201 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc
     7261 caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga ggatcgtttc
     7321 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     7381 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     7441 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     7501 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     7561 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     7621 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     7681 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     7741 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     7801 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     7861 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     7921 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     7981 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     8041 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     8101 ttgacgagtt cttctgaatt attaacgctt acaatttcct gatgcggtat tttctcctta
     8161 cgcatctgtg cggtatttca caccgcatca ggtggcactt ttcggggaaa tgtgcgcgga
     8221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagattatca
     8281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     8341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     8401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     8461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     9001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     9061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     9121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     9181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     9241 caatattatt gaagcattta tcagggttat tgtctcatga ccaaaatccc ttaacgtgag
     9301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     9361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     9421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     9481 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     9541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     9601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     9661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     9721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     9781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     9841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     9901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     9961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
    10021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
    10081 acgaccgagc gcagcgag
//