pMGS46 (ARF16-PB1-HA-P2A-OsTIR1)

LOCUS       pMGS46 (ARF16-PB1-HA-P2A-OsTIR1)              10247 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  A repair construct to express ARF16-PB1-HA and OsTIR1 under the 
            control of the CMV promoter from the human AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    pMGS46 (ARF16-PB1-HA-P2A-OsTIR1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10247)
  AUTHORS   Sathyan KM, McKenna BD, Anderson WD, Duarte FM, Core L, Guertin MJ
  TITLE     An improved auxin-inducible degron system preserves native protein 
            levels and enables rapid and specific protein depletion.
  JOURNAL   Genes Dev. 2019 Oct 1;33(19-20):1441-1455. doi: 
            10.1101/gad.328237.119. Epub 2019 Aug 29.
  PUBMED    31467088
REFERENCE   2  (bases 1 to 10247)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10247
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    295..1098
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     enhancer        1155..1458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1459..1662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1613..1633
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1703..1723
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2261..2287
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2297..2353
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2363..4090
                     /codon_start=1
                     /product="rice transport inhibitor response 1 protein"
                     /label=OsTIR1
                     /note="F-box protein that mediates auxin-dependent 
                     interaction of a ubiquitin ligase with an auxin-inducible 
                     degron (Tan et al., 2007)"
                     /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
                     GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
                     LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
                     EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
                     AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
                     LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
                     VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
                     CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
                     AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
                     CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
     polyA_signal    4264..4385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4301..4320)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4355..4374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4394..4894
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4813..4831
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             4916..5515
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4916..4935)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5412..5432
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5522..5746
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    5749..6585
                     /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(6619..6638)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6620..6638)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6645..6662)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6645..6661)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6654..6676)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             6799..7101
                     /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     6980..6999
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             7450..8244
                     /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(7504..7523)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8114..8133
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(8494..9354)
                     /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     9117..9136
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9478..10066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9967..9986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10220..10237
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt
      301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg
      361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct
      421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc
      481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg
      541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc
      601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg
      661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt
      721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt
      781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg
      841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca
      901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat
      961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc
     1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc
     1081 tccaccccac agtggggcaa ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     1141 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1201 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1261 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1321 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1381 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1441 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     1501 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     1561 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     1621 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     1681 atccgctagc gctgccacca tgcctaagaa gaagcggaag gtgggcagca cactgacttc
     1741 cgggcagcgg gatcaccggg acgtcgagaa cgagctgagc tccgccgcct tcagctccca
     1801 gagcttcggg gtgcccgaca tgagctttaa gcccggctgc agcagcgacg tggccgtgac
     1861 cgacgccggg atgcccagcc aggggctgtg gaacaaccag acccagcgga tgaggacatt
     1921 cacaaaggtg cagaagcggg ggagcgtggg ccggagcatc gatatcacca ggtacaggga
     1981 ttacgatgag ctgaggcatg atctcgcctg tatgttcggc atccagggcc agctggagga
     2041 cccctaccgg atggactgga agctggtgta cgtggaccac gagaacgata tcctgctggt
     2101 gggcgatgac ccctgggagg agttcgtggg ctgcgtgaag agcattaaga ttctgtctgc
     2161 tgctgaggtg cagcagatga gcctggatgg agatctggga ggagtgccac ctcagactca
     2221 ggcctgtagt gctagtgatg atgctaacgc ctggagaggg tatccatatg acgtcccaga
     2281 ctatgccgga agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga
     2341 gaaccctgga cctgctgcca ccatgacata ctttcctgaa gaggtcgtcg aacacatttt
     2401 tagcttcctg cctgcacaga gagatagaaa cacagtgagc ctggtctgca aagtgtggta
     2461 cgagatcgaa cgcctgagcc ggagaggagt gttcgtcggc aactgctatg ctgtgagagc
     2521 aggcagggtc gccgctaggt ttccaaatgt gcgcgcactg accgtcaagg ggaaacccca
     2581 cttcgccgac tttaacctgg tgccccctga ttggggagga tacgccggcc cttggatcga
     2641 ggcagccgct cgcggctgtc atggactgga ggaactgcgc atgaagcgaa tggtggtctc
     2701 tgacgaaagt ctggagctgc tggctcggag cttccctagg tttcgcgcac tggtgctgat
     2761 ttcttgcgaa ggcttcagca ccgatggact ggcagccgtg gcctcccact gtaagctgct
     2821 gcgggagctg gacctccagg agaatgaagt ggaggataga ggccccagat ggctgtcttg
     2881 cttcccagac tcatgtacca gcctggtgtc cctgaacttt gcctgcatca aaggcgaagt
     2941 gaatgctggg tccctggagc ggctggtctc aagaagcccc aacctgaggt ctctgcggct
     3001 gaaccggagc gtgagcgtgg acactctggc taagattctg ctgagaaccc ctaacctgga
     3061 ggatctggga accggcaatc tgacagacga tttccagaca gaatcctact ttaaactgac
     3121 ttctgccctg gagaagtgta aaatgctgag gagtctgtca ggattctggg atgcttcacc
     3181 cgtgtgcctg agctttatct accctctgtg tgcacagctg acaggcctga acctgagcta
     3241 tgcaccaacc ctggacgcca gtgatctgac aaagatgatc tcacgctgcg tgaaactcca
     3301 gcgactgtgg gtgctggact gtatttccga taaggggctc caggtggtcg ccagctcctg
     3361 caaggacctc caggagctga gagtgttccc atctgatttt tacgtggccg gatatagtgc
     3421 tgtcactgag gaaggcctgg tggcagtctc actgggatgc ccaaagctga acagcctgct
     3481 gtatttctgt catcagatga ctaatgctgc actggtgacc gtcgccaaga actgccctaa
     3541 tttcacccga tttcggctgt gtattctgga accaggcaaa cccgacgtgg tcacatccca
     3601 gccactggat gaagggtttg gagctatcgt gagagagtgc aagggactcc agaggctgag
     3661 catttccggc ctgctgacag acaaagtgtt catgtacatc ggcaagtatg ctaagcagct
     3721 ggagatgctg agcattgcat ttgccggaga ctccgataag ggcatgatgc acgtgatgaa
     3781 cgggtgtaag aatctgcgaa aactggaaat ccgggacagc cctttcgggg atgccgctct
     3841 gctgggaaac tttgccagat acgagacaat gaggagcctg tggatgtcta gttgcaatgt
     3901 gactctgaag ggctgtcagg tcctggctag taaaatgcct atgctgaacg tggaagtcat
     3961 taatgagcgg gacgggtcta acgaaatgga ggaaaatcat ggcgacctgc caaaggtgga
     4021 gaaactgtat gtgtatcgga ccaccgcagg ggcaagagat gatgctccca actttgtgaa
     4081 gattctgtga ggatccatcg gatcccgggc ccgtcgacgg taccgcgggc ccgggatcca
     4141 ccggggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     4201 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4261 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4321 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4381 tcttaagttg gggttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg
     4441 ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac
     4501 gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct
     4561 tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca
     4621 aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc
     4681 agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc gcgccgagag
     4741 cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt
     4801 cctgcccgcg cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc
     4861 cctcgttgac cgaatcaccg acctctctcc ccagggggat ccaccggagc ttaccatgac
     4921 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     4981 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     5041 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     5101 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     5161 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     5221 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     5281 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     5341 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     5401 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     5461 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaggtac
     5521 cctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5581 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5641 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5701 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggttac tagggacagg
     5761 attggtgaca gaaaagcccc atccttaggc ctcctccttc ctagtctcct gatattgggt
     5821 ctaaccccca cctcctgtta ggcagattcc ttatctggtg acacaccccc atttcctgga
     5881 gccatctctc tccttgccag aacctctaag gtttgcttac gatggagcca gagaggatcc
     5941 tgggagggag agcttggcag ggggtgggag ggaagggggg gatgcgtgac ctgcccggtt
     6001 ctcagtggcc accctgcgct accctctccc agaacctgag ctgctctgac gcggctgtct
     6061 ggtgcgtttc actgatcctg gtgctgcagc ttccttacac ttcccaagag gagaagcagt
     6121 ttggaaaaac aaaatcagaa taagttggtc ctgagttcta actttggctc ttcacctttc
     6181 tagtccccaa tttatattgt tcctccgtgc gtcagtttta cctgtgagat aaggccagta
     6241 gccagccccg tcctggcagg gctgtggtga ggaggggggt gtccgtgtgg aaaactccct
     6301 ttgtgagaat ggtgcgtcct aggtgttcac caggtcgtgg ccgcctctac tccctttctc
     6361 tttctccatc cttctttcct taaagagtcc ccagtgctat ctgggacata ttcctccgcc
     6421 cagagcaggg tcccgcttcc ctaaggccct gctctgggct tctgggtttg agtccttggc
     6481 aagcccagga gaggcgctca ggcttccctg tcccccttcc tcgtccacca tctcatgccc
     6541 ctggctctcc tgccccttcc ctacaggggt tcctggctct gctctaaggg cgaattcgcg
     6601 gccgctaaat tcaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
     6661 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     6721 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     6781 cgcagcctat acgtacggca gtttaaggtt tacacctata aaagagagag ccgttatcgt
     6841 ctgtttgtgg atgtacagag tgatattatt gacacgccgg ggcgacggat ggtgatcccc
     6901 ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat
     6961 atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt
     7021 atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac
     7081 ctgatgttct ggggaatata aatgtcaggc atgagattat caaaaaggat cttcacctag
     7141 atccttttca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
     7201 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
     7261 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
     7321 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     7381 ttgccgccaa ggatctgatg gcgcagggga tcaagctctg atcaagagac aggatgagga
     7441 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     7501 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     7561 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     7621 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     7681 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     7741 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     7801 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     7861 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     7921 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     7981 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     8041 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     8101 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     8161 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     8221 cgccttcttg acgagttctt ctgaattatt aacgcttaca atttcctgat gcggtatttt
     8281 ctccttacgc atctgtgcgg tatttcacac cgcatcaggt ggcacttttc ggggaaatgt
     8341 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     8401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgacca aaatccctta
     9421 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     9481 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     9541 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     9601 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     9661 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     9721 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     9781 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     9841 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     9901 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     9961 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
    10021 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
    10081 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
    10141 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
    10201 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//