LOCUS       sequence-360358-       10173 bp    DNA     circular SYN 15-APR-2025
DEFINITION  sgRNA targeting rat PKA-RII-alpha subunits.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10173)
  AUTHORS   Church TW, Tewatia P, Hannan S, Antunes J, Eriksson O, Smart TG, 
            Hellgren Kotaleski J, Gold MG
  TITLE     AKAP79 enables calcineurin to directly suppress protein kinase A 
            activity.
  JOURNAL   Elife. 2021 Oct 6;10. pii: 68164. doi: 10.7554/eLife.68164.
  PUBMED    34612814
REFERENCE   2  (bases 1 to 10173)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2021-10-6"; volume: "10. pii: 68164. doi"; pages: " 
            10.7554/eLife.68164"
COMMENT     Sequence Label: pFUGW-shRII-alpha
FEATURES             Location/Qualifiers
     source          1..10173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(192..211)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     539..560
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    632..1220
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(685..705)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1103..1114)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1223..1239)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1224..1240)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             1745..1925
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(1942..1959)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1948..2172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2218..2646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2305..2324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2515..2536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2655..2675)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2660..2989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2840..2975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2902..2921
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3037..3084
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3103..3477
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3607..3728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3644..3663)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3698..3717
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3777..3793)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3777..3793)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3790..3812)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3801..3817
                     /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)."
     promoter        complement(join(3825..3831,3832..3849,3850..3855))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3870..3891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4008..4025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4179..4767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4259..4278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4938..5729,5730..5798))
                     /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     5561..5580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5799..5903)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5978..5997)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6169..6548
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6550..6748
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6703..6723
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6766..6946
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6993..7118
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7611..7844
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8029..8073
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8371..8488
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(8636..8850)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(8696..8715)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        8879..9278
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     9927..9946
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      10053..10062
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(10059..10061,10062..10064,10065..10173,1..605)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(10059..10061,10062..10064,10065..10173,1..602)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(10104..10125)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
       61 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      121 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      181 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      241 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      301 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
      361 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
      421 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
      481 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
      541 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
      601 agtaaagcaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
      661 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
      721 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
      781 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
      841 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
      901 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
      961 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     1021 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     1081 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     1141 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     1201 cctttgggcc gcctccccgc atcgataccg tcgacctcga gacctagaaa aacatggagc
     1261 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     1321 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     1381 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     1441 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     1501 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
     1561 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
     1621 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
     1681 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
     1741 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     1801 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     1861 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     1921 tagcagaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1981 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2041 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2101 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2161 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2221 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2281 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2341 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2401 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2461 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2521 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2581 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2641 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2701 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2761 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2821 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2881 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     2941 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3001 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     3061 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     3121 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     3181 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     3241 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     3301 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     3361 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     3421 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     3481 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3541 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3601 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3661 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3721 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     3781 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3841 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3901 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3961 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4021 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4081 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4141 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4201 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4261 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4321 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4381 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4441 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4501 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4561 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4621 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4681 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4741 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4801 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4861 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4921 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4981 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5041 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5101 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5161 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5221 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5281 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5341 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5401 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5461 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5521 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5581 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5641 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5701 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5761 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5821 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5881 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     5941 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     6001 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     6061 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     6121 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
     6181 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     6241 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     6301 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     6361 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6421 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6481 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     6541 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     6601 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     6661 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     6721 gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta
     6781 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6841 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6901 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     6961 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     7021 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     7081 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     7141 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     7201 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     7261 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     7321 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     7381 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     7441 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     7501 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     7561 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     7621 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     7681 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     7741 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     7801 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     7861 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     7921 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     7981 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     8041 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     8101 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     8161 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     8221 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     8281 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     8341 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     8401 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     8461 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     8521 ttggttaatt aagaaaggat ccgaaagaat tcgcccttag atctctagac aaaaaagcca
     8581 ggaatcagac tcgttcatct cttgaatgaa cgagtctgat tcctggcggg aaagagtggt
     8641 ctcatacaga acttataaga ttcccaaatc caaagacatt tcacgtttat ggtgatttcc
     8701 cagaacacat agcgacatgc aaatattgca gggcgccact cccctgtccc tcacagccat
     8761 cttcctgcca gggcgcacgc gcgctgggtg ttcccgccta gtgacactgg gcccgcgatt
     8821 ccttggagcg ggttgatgac gtcagcgttc ttaattaacc cgtgtcggct ccagatctgg
     8881 cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc
     8941 acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct caggacagcg
     9001 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
     9061 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
     9121 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
     9181 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg
     9241 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctag
     9301 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca
     9361 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca
     9421 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg
     9481 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gaggccttcg
     9541 ctaatgcggg aaagctctta ttcgggtgag atgggctggg gcaccatctg gggaccctga
     9601 cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta
     9661 tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt
     9721 gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg taggtgtgcg
     9781 gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg
     9841 acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc tttggtcggt
     9901 tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc gctcggggtt
     9961 ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat ttgggtcaat
    10021 atgtaatttt cagtgttaga ctagtaccgg tcgccaccat ggtgagcaag ggcgaggagc
    10081 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
    10141 tcagcgtgtc cggcgagggc gagggcgatg cca
//
