LOCUS       pAAV-CAG-RiboL1-jGCaMP8 6740 bp ds-DNA     circular     27-SEP-2022
DEFINITION  .
KEYWORDS    "accession:addgene_59462_92647"
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
COMMENT     GENERAL Depositing Lab: Edward Boyden Publication: Accessory
            plasmids for optogenetics (unpublished) Purpose: AAV-mediated
            expression of tdTomato (codon diversified) under the CAG promoter
            BACKBONE Backbone manufacturer: Scott Sternson Backbone size w/o
            insert (bp): 4719 Total vector size (bp): 6150 Vector backbone: AAV
            with CAG promter Vector type: Mammalian Expression, AAV CLONING
            INFORMATION 3′ cloning site: EcoRI (not destroyed) 3′ sequencing
            primer: gcagcgtatccacatagcg 5′ cloning site: BamHI (not destroyed)
            5′ sequencing primer: gctctagagcctctgctaacc Cloning method:
            Restriction Enzyme INFORMATION FOR AAV RETROGRADE (CATALOG #
            59462-AAVRG) ( BACK TO TOP ) Buffer: PBS + 0.001% Pluronic F-68 +
            200 mM NaCl Packaging Plasmids: encode adenoviral helper sequences
            and AAV Pricing: $350 USD for preparation of 100 µL virus + $30 USD
            for plasmid. Purification: Iodixanol gradient ultracentrifugation
            Reporter Gene: tdTomato Serotype:  Shipment: Viral particles are
            shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be
            included in the shipment. Storage: Store at -80℃. Thaw just before
            use and keep on ice. Terms and Licenses:  Titer: ≥ 7×10¹² vg/mL
            Volume: 100 µL INFORMATION FOR AAV5 (CATALOG # 59462-AAV5) ( BACK
            TO TOP ) Buffer: PBS + 0.001% Pluronic F-68 Packaging Plasmids:
            encode adenoviral helper sequences and AAV Pricing: $350 USD for
            preparation of 100 µL virus + $30 USD for plasmid. Purification:
            Iodixanol gradient ultracentrifugation Reporter Gene: tdTomato
            (codon diversified) Serotype: AAV5 Shipment: Viral particles are
            shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be
            included in the shipment. Storage: Store at -80℃. Thaw just before
            use and keep on ice. Terms and Licenses:  Titer: ≥ 5×10¹² vg/mL
            Volume: 100 µL INFORMATION FOR AAV PHP.S (CATALOG # 59462-PHP.S) (
            BACK TO TOP ) Buffer: PBS + 0.001% Pluronic F-68 Packaging
            Plasmids: encode adenoviral helper sequences and AAV Pricing: $350
            USD for preparation of 100 µL virus + $30 USD for plasmid.
            Purification: Iodixanol gradient ultracentrifugation Reporter Gene:
            tdTomato Serotype:  Shipment: Viral particles are shipped frozen on
            dry ice. Plasmid DNA (≥ 200ng) will also be included in the
            shipment. Storage: Store at -80℃. Thaw just before use and keep on
            ice. Terms and Licenses:  Titer: ≥ 1×10¹³ vg/mL Volume: 100 µL
            INFORMATION FOR AAV2 (CATALOG # 59462-AAV2) ( BACK TO TOP ) Buffer:
            PBS + 0.001% Pluronic F-68 Packaging Plasmids: encode adenoviral
            helper sequences and AAV Pricing: $350 USD for preparation of 100
            µL virus + $30 USD for plasmid. Purification: Iodixanol gradient
            ultracentrifugation Reporter Gene: tdTomato (codon diversified)
            Serotype: AAV2 Shipment: Viral particles are shipped frozen on dry
            ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.
            Storage: Store at -80℃. Thaw just before use and keep on ice. Terms
            and Licenses:  Titer: ≥ 7×10¹² vg/mL Volume: 100 µL GROWTH IN
            BACTERIA Bacterial Resistance(s): Ampicillin Copy number: Low Copy
            Growth Strain(s): NEB Stable Growth Temperature: 37°C Growth
            instructions: Stbl3 at 30C (use carbenicillin if using Stbl3) OR
            DH5a at 37C GENE/INSERT Gene/Insert name: tdTomato Insert Size
            (bp): 1431 Mutation: codon diversified Promoter: CAG Species:
            Synthetic INFORMATION FOR AAV PHP.EB (CATALOG # 59462-PHPEB) ( BACK
            TO TOP ) Buffer: PBS + 0.001% Pluronic F-68 Packaging Plasmids:
            encode adenoviral helper sequences and AAV Pricing: $350 USD for
            preparation of 100 µL virus + $30 USD for plasmid. Purification:
            Iodixanol gradient ultracentrifugation Reporter Gene: tdTomato
            Serotype:  Shipment: Viral particles are shipped frozen on dry ice.
            Plasmid DNA (≥ 200ng) will also be included in the shipment.
            Storage: Store at -80℃. Thaw just before use and keep on ice. Terms
            and Licenses:  Titer: ≥ 1×10¹³ vg/mL Volume: 100 µL INFORMATION FOR
            AAV1 (CATALOG # 59462-AAV1) ( BACK TO TOP ) Buffer: PBS + 0.001%
            Pluronic F-68 Packaging Plasmids: encode adenoviral helper
            sequences and AAV Pricing: $350 USD for preparation of 100 µL virus
            + $30 USD for plasmid. Purification: Iodixanol gradient
            ultracentrifugation Reporter Gene: tdTomato Serotype: AAV1
            Shipment: Viral particles are shipped frozen on dry ice. Plasmid
            DNA (≥ 200ng) will also be included in the shipment. Storage: Store
            at -80℃. Thaw just before use and keep on ice. Terms and Licenses:
            Titer: ≥ 1×10¹³ vg/mL Volume: 100 µL
FEATURES             Location/Qualifiers
     primer_bind     complement(11..28)
                     /label="L4440"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     rep_origin      complement(182..770)
                     /label="ori"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     complement(262..281)
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             complement(941..1801)
                     /label="AmpR"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer_bind     1564..1583
                     /label="Amp-R"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     promoter        complement(1802..1906)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     rep_origin      complement(1932..2387)
                     /label="f1 ori"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer_bind     complement(2069..2090)
                     /label="F1ori-F"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer_bind     2281..2300
                     /label="F1ori-R"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     repeat_region   2459..2588
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          complement(2471..2486)
                     /label="297_AAVtitering_qPCR_ITR_R"
                     /note="sequence: CGGCCTCAGTGAGCGA"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          2526..2541
                     /label="297_AAVtitering_qPCR_ITR_R"
                     /note="sequence: CGGCCTCAGTGAGCGA"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(2567..2587)
                     /label="296_AAVtitering_qPCR_ITR_F"
                     /note="sequence: GGAACCCCTAGTGATGGAGTT"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     promoter        2901..3178
                     /label="chicken beta-actin promoter"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     3458..3480
                     /label="pCAGGS-5"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer_bind     3542..3561
                     /label="pCAG-F"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          3595..3613
                     /label="779_nhel-RiboL1-F"
                     /note="sequence: TAAGCAGCTAGCCCGCCACCATGCATCATC"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          3598..3615
                     /label="l1i8m-tre-gibson2021-F"
                     /note="sequence: acttcctaccctcgtaaagtGCCACCATGCATCATCAC"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    3604..4869
                     /label="jGCaMP8s"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             3604..5601
                     /label="Translation 3604-5601"
     CDS             3604..5601
                     /label="Translation 3604-5601"
     primer          4845..4869
                     /label="149_pAAV-hSyn1-Flex-mRuby2-GSG-P2A-GCaMP6s-WPRE-pA_seq_R"
                     /note="sequence: gtttgtacaaatgatgacagcgaag"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    4870..4950
                     /label="Linker"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             4951..5601
                     /label="RL10"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          complement(5591..5610)
                     /label="l18m-tre-gibson2021-R"
                     /note="sequence: ataagcttgatatcgaattgggcgcgccctaatacagac"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          complement(5592..5609)
                     /label="780_RiboL1-R"
                     /note="sequence: TACTATGGCGCGCCCTAATACAGA"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    5642..6230
                     /label="WPRE"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer_bind     complement(5695..5715)
                     /label="WPRE-R"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          5997..6016
                     /label="298_WPRE-forward"
                     /note="sequence: GGCTGTTGGGCACTGACAAT"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(6097..6116)
                     /label="299_WPRE-reverse"
                     /note="sequence: CCGAAGGGACGTAGCAGAAG"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             complement(6113..6124)
                     /label="Factor Xa site"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     polyA_signal    6273..6394
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer_bind     complement(6284..6303)
                     /label="EBV-rev"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer_bind     6338..6357
                     /label="SV40pA-R"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     repeat_region   6556..6685
                     /label="AAV2 ITR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          6557..6577
                     /label="296_AAVtitering_qPCR_ITR_F"
                     /note="sequence: GGAACCCCTAGTGATGGAGTT"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(6603..6618)
                     /label="297_AAVtitering_qPCR_ITR_R"
                     /note="sequence: CGGCCTCAGTGAGCGA"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          6658..6673
                     /label="297_AAVtitering_qPCR_ITR_R"
                     /note="sequence: CGGCCTCAGTGAGCGA"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     2701 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2761 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2821 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2881 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
     2941 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
     3001 gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
     3061 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
     3121 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
     3181 agtcgctgcg acgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
     3241 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
     3301 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
     3361 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg
     3421 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     3481 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     3541 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgGA TCCACCCGCC
     3601 ACCATGCATC ATCACCATCA TCACACGCGT CGCAAGAAGA CCTTCAAGGA GGTGGCCACC
     3661 GCCGTGAAGA TCATCGCCAT GCTGATGGGC CTGAAGATCA ACGTCTATAT CAAGGCCGAC
     3721 AAGCAGAAGA ACGGCATCAA GGCGAACTTC CACATCCGCC ACAACATCGA GGACGGCGGC
     3781 GTGCAGCTCG CCTACCACTA CCAGCAGAAC ACCCCCATCG GCGACGGCCC CGTGCTGCTG
     3841 CCCGACAACC ACTACCTGAG CGTGGAGTCC AAACTTTCGA AAGACCCCAA CGAGAAGCGC
     3901 GATCACATGG TCCTGCTGGA GTTCGTGACC GCCGCCGGGA TCACTCTCGG CATGGACGAG
     3961 CTGTACAAGG GCGGTACCGG AGGGAGCATG GTGAGCAAGG GCGAGGAGCT GTTCACCGGG
     4021 GTGGTGCCCA TCCTGGTCGA GCTGGACGGC GACGTAAACG GCCACAAGTT CAGCGTGTCC
     4081 GGCGAGGGTG AGGGCGATGC CACCTACGGC AAGCTGACCC TGAAGTTCAT CTGCACCACC
     4141 GGCAAGCTGC CCGTGCCCTG GCCCACCCTC GTGACCACCC TGACCTACGG CGTGCAGTGC
     4201 TTCAGCCGCT ACCCCGACCA CATGAAGCAG CACGACTTCT TCAAGTCCGC CATGCCCGAA
     4261 GGCTACATCC AGGAGCGCAC CATCTTCTTC AAGGACGACG GCAACTACAA GACCCGCGCC
     4321 GAGGTGAAGT TCGAGGGCGA CACCCTGGTG AACCGCATCG AGCTGAAGGG CATCGACTTC
     4381 AAGGAGGACG GCAACATCCT GGGGCACAAG CTGGAGTACA ACCTGCCGGA CCAACTGACT
     4441 GAAGAGCAGA TCGCAGAATA CAAAGAGGCT TTCTCCCTAT TTGACAAGGA CGGGGATGGG
     4501 ACAATAACAA CCAAGGAGCT GGGGACGGTG ATGCGGTCTC TCGGACACAA CCCCACAGAA
     4561 GCAGAGCTGC AGGACATGAT CAATGAAGTA GATGCCGACG GTGACGGCAC AATCGACTTC
     4621 CCTGAGTTCC TGACAATGAT GGCAAGAAAA ATGAAATACA GGGACACGGA AGAAGAAATT
     4681 AGAGAAGCGT TCGGTGTGTT TGATAAGGAT GGCAATGGCT ACATCAGTGC AGCAGAGCTT
     4741 CGCCACGTGA TGACAAACCT TGGAGAGAAG TTaacagatg aagaggttga tgaaatgatc
     4801 agggaagcag acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg
     4861 acagcgaagG GGGGCAGCGG TGGGTCTGGA GGGACTGGGG GTAGCGGTGG TAGCGGTGGG
     4921 ACAGGGGGAA GCGGCGGTAG TGGTGGAACA agcagcaaag tctcacgcga caccctgtac
     4981 gaggcggtgc gggaagtcct gcacgggaac cagcgcaagc gccgcaagtt tctggagacg
     5041 gtggagctgc agatcagcct gaagaactac gaccctcaga aggacaaacg tttctcgggc
     5101 accgtcaggc tcaagtccac cccacgcccc aagttctcgg tgtgcgttct gggggaccag
     5161 cagcactgtg atgaagccaa ggccgtggat atcccccaca tggacatcga ggcgctcaag
     5221 aaacttaaca aaaacaagaa gttggtcaag aagctggcta agaagtacga tgcctttttg
     5281 gcctctgagt ctctgattaa gcagatccca cgtatcctgg gcccaggcct aaacaaggct
     5341 ggcaagttcc cctccctgct gacacacaat gaaaacatgg tggccaaagt ggatgaggtg
     5401 aaatcgacaa tcaagttcca gatgaagaag gtgctgtgtt tggccgtcgc tgttggccac
     5461 gtgaagatga ccgatgatga gctagtctac aacattcatc tggctgtcaa tttcttggtg
     5521 tccttgctta agaaaaactg gcaaaacgtg cgggctctgt acatcaagag caccatgggc
     5581 aagccccagc gtctgtatta gggcgcgccc ctgcagggaa ttcgatatca agcttatcga
     5641 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5701 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     5761 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     5821 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     5881 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     5941 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6001 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     6061 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6121 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6181 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     6241 tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     6301 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     6361 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     6421 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     6481 tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt
     6541 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     6601 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     6661 cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg
     6721 cggtttgcgt attgggcgct
//