LOCUS       sequence-343660-       16379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of RVdG-CVS-N2c viral vectors for cell-type specific 
            trans-synaptic retrograde labeling..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16379)
  AUTHORS   Sumser A, Joesch M, Jonas P, Ben-Simon Y
  TITLE     Fast, high-throughput production of improved rabies viral vectors 
            for specific, efficient and versatile transsynaptic retrograde 
            labeling.
  JOURNAL   Elife. 2022 Aug 30;11:e79848. doi: 10.7554/eLife.79848.
  PUBMED    36040301
REFERENCE   2  (bases 1 to 16379)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2022-08-30"; volume: "11:e79848. doi"; pages: " 
            10.7554/eLife.79848"
COMMENT     Sequence Label: CVS-N2c-nl.EGFP-jRGECO1a
FEATURES             Location/Qualifiers
     source          1..16379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3319..3328
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3325..3327,3328..3330,3331..4041)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3325..3327,3328..3330,3331..4041)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3370..3391)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3631..3650)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3978..3999
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4054..4074
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4078..4098
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4102..4122
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4292..4309
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4313..4345
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(4349..4357,4358..4372)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             4400..4456
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     5152..5171
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7808..7819
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(9611..9622)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12818..12836)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      12832..12879
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(12907..12923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12907..12923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12920..12942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12931..12947
                     /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(12955..12961,12962..12979,12980..12985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13000..13021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13138..13155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13309..13897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13389..13408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14068..14859,14860..14928))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGASKGGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     14691..14710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14929..15033)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      15060..15516
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(15147..15166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15357..15378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15642..15664
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15656..15673
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15657..15673
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        15743..16046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        16047..16250
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     16201..16221
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     16295..16314
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        16295..16313
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acgcttaaca acaaaaccag agaagaaaaa gacagcgtca attgcaaagc aaaaatgtaa
       61 cacccctaca atggatgccg acaagattgt gttcaaagtc aataatcagg tggtctcttt
      121 gaagcctgag attatcgtgg atcaatatga gtacaagtac cctgccatca aggatttgaa
      181 aaagccttgt atcaccctag ggaaagcccc cgacttgaac aaagcataca aatcagtttt
      241 atcaggcatg aatgccgcca aacttgatcc ggatgatgta tgctcctact tggcagcagc
      301 aatgcagttc tttgagggga catgtccgga agactggacc agctatggaa tcctgattgc
      361 acgaaaagga gataggatca ccccaaactc tctagtggag ataaagcgta ctgatgtaga
      421 agggaattgg gctctgacag gaggcatgga attgacaagg gaccccactg tctctgaaca
      481 tgcatcttta gtcggtcttc tcctgagtct gtacaggttg agcaaaatat caggacagaa
      541 cactggtaac tataagacaa acattgcaga taggatagag cagattttcg agacagcacc
      601 ttttgttaag atcgtggaac accataccct aatgacaact cacaagatgt gtgctaattg
      661 gagtactata ccgaacttca gatttttggc cggaacctac gacatgtttt tctcacggat
      721 tgagcatcta tattcggcaa tcagagtggg cacagtcgtc accgcttatg aagactgctc
      781 aggactggta tcgtttacag ggttcataaa gcagatcaat ctcaccgcaa gggaagcaat
      841 actatatttc ttccacaaga actttgagga agagataaga agaatgttcg agccagggca
      901 agagacagct gttcctcact cttatttcat ccacttccgt tcactaggct tgagtgggaa
      961 gtctccttat tcatcgaatg ctgtcggtca tgtgttcaat ctcattcact ttgttggatg
     1021 ctacatgggt caagtcagat ctctaaatgc gacggttatt gctgcatgtg cccctcatga
     1081 gatgtctgtt ctagggggct atttgggaga ggaattcttc ggaaaaggga catttgaaag
     1141 aaggttcttc agagacgaga aagaacttca agaatatgag gcggctgaac taacaaagtc
     1201 cgacgtggca ctggcagatg acggaaccgt caactctgat gacgaggact atttctctgg
     1261 tgaaaccaga agtccagaag ctgtctatac tcgaatcatg atgaatggag gtcgactgaa
     1321 gagatctcat atacggagat atgtctcagt cagttccaat catcaagccc gtccaaactc
     1381 attcgccgaa tttttaaaca agacgtattc gaatgactca taaggagttg attgacaggg
     1441 tgccagaaat ctatagattg tatatatcca tcatgaaaaa aactaacact cctcctttca
     1501 aaccatccca aatatgagca agatctttgt taatccgagt gcaatcagag ccggtctggc
     1561 cgatcttgag atggccgaag agactgttga tctgatcaac agaaacatag aagacaatca
     1621 ggctcatctc cagggagaac ccatagaagt ggacaacctc cctgaggaca tgaagcgact
     1681 tcacctggac gatgaaaaat cgtccaacct tggtgagatg gttagggtgg gagaaggcaa
     1741 gtatcgagag gactttcaga tggatgaggg agaggacccc aacctcctgt tccaatcgta
     1801 cctggataat gttggagtcc aaatagtcag acaaatgagg tcaggagaga gattcctcaa
     1861 gatatggtca cagaccgtag aggaaattgt atcctatgtc acggtcaact ttcctaaccc
     1921 tccaagaagg tcttcggagg ataaatcaac ccagactact ggcagagagc tcaagaagga
     1981 gacaacgtct gctttctctc agagagaaag ccaaccttcg aaagctagga tggtggctca
     2041 agttgcccct ggtcctccag cccttgaatg gtcagccacc aatgaagaag atgatctatc
     2101 agtagaggct gagatcgctc atcagattgc tgaaagcttt tccaagaagt acaagtttcc
     2161 ctcccgatct tcaggaatat tcttgtataa ttttgagcaa ctgaagatga accttgatga
     2221 catagttaaa gaggcaaaaa atgtaccggg cgtgacccgt ctggcccatg atggatccaa
     2281 aatccccctg agatgcgtac tgggatgggt cgctttggct aattccaaaa aattccaatt
     2341 actagtcgag gctgacaagc taagcaaaat catgcaagat gatttgaatc gctatacatc
     2401 ctgctaaccg agttttcgaa ctcagtccct ccagataatg aaaactgaga tgttatggag
     2461 tagacatgaa aaaaacaggc aacaccactg ataaaatgaa cgttctacgc aagatagtga
     2521 aaaaatgtag ggatgaggac actcaaaagc cctctcctgt gtcagcccct ccgtatgacg
     2581 atgacctgtg gcttccacct cctgaatatg tcccgctgaa agaactcaca agcaagaaga
     2641 acatgaggaa cttttgtgtc aacggggagg ttaaagcgtg tagcccaaat ggttactcat
     2701 tcaggatttt gcggcacatt ctgagatcat tcaacgagat atactctggg aatcatagga
     2761 tgattgggtt agtcaaaatt gttgttggac tagctttatc aggagctcca gtacctgagg
     2821 gcatgaactg ggtatacaaa ttgaggagaa cccttatatt ccagtgggct gattccaggg
     2881 gccctcttga aggggaggag ttagattact ctcaagagat cacttgggat gatgatactg
     2941 aattcgtcgg attgcaaata agagtgagcg caagacaatg tcatattcaa ggcaggatct
     3001 ggtgtatcaa cacgaactcg agggcatgtc aactatggtc tgacatgtct cttcagacac
     3061 aaaggtctga agaggacaaa gactcttctc tgcttctaga ataatcagat tatatcctgc
     3121 aagtgtatca cttgtttacc tctggaggag agagcataca ggcttgactc cgatccttgg
     3181 gagcaataga acaaaaaaac acacgttatg gtgccgttaa atcgctgcat tttatcaaag
     3241 tcaagttgat aacctttaca ttttgagcct cttggatgtg aaaaaaacta ttaacatccc
     3301 tcaaaagacc ccggtaaagc caccatggtg agcaagggcg aggagctgtt caccggggtg
     3361 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     3421 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3481 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     3541 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     3601 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3661 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3721 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3781 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3841 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3901 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3961 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     4021 ggcatggacg agctgtacaa ggctcgagct gatccaaaaa agaagagaaa ggtagatcca
     4081 aaaaagaaga gaaaggtaga tccaaaaaag aagagaaagg taggatccag ataactgatc
     4141 ataatcagcc ataccacatt tggaattcgt cgagggggcg cgccgctaga catgaaaaaa
     4201 actaacactc ctccggtacg ccaccatgct gcagaacgag cttgctctta agttggctgg
     4261 acttgatatt aacaagactg gaggaggttc tcatcatcat catcatcatg gtatggctag
     4321 catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc
     4381 aacaatggtc gactcatcac gtcgtaagtg gaataaggca ggtcacgcag tcagagctat
     4441 aggtcggctg agctcacccg tggtttccga gcggatgtac cccgaggacg gcgccctgaa
     4501 gagcgagatc aagaaggggc tgaggctgaa ggacggcggc cactacgccg ccgaggtcaa
     4561 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacatcg tggacatcaa
     4621 gttggacatc gtgtcccaca acgaggacta caccatcgtg gaacagtgcg aacgcgccga
     4681 gggccgccac tccaccggcg gcatggacga gctgtacaag ggaggtacag gcgggagtct
     4741 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     4801 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     4861 cccctacgag gcctttcaga ccgctaagct gaaggtgacc aagggtggcc ccctgccctt
     4921 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctaca ttaagcaccc
     4981 agccgacatc cccgactact tcaagctgtc cttccccgag ggcttcaggt gggagcgcgt
     5041 gatgaacttc gaggacggcg gcattattca cgtgaaccag gactcctccc tgcaggacgg
     5101 cgtattcatc tacaaggtga agctgcgcgg caccaacttc ccccccgacg gccccgtaat
     5161 gcagaagaag accatgggct gggaggctac gcgtgacgac ctgactgaag agcagatcgc
     5221 agaatttaaa gaggctttct ccctatttga caaggacggg gatgggacga taacaaccaa
     5281 ggagcttggg acggtgttcc gatcgctggg gcagaacccc acagaagcag agctgcagga
     5341 catgatcaat gaagtagatg ccgacggtga cggcacattc gacttccctg agttcctgac
     5401 gatgatggca agaaaaatga atgacacaga cagtgaagag gaaattcgcg aagcgttccg
     5461 cgtgtttgat aaggacggca atggctacat cggcgcagca gagcttcgcc acgtgatgac
     5521 agaccttgga gagaagttaa cagatgagga ggttgatgaa atgatcaggg tagcagacat
     5581 cgatggggat ggtcaggtaa actacgaaga gtttgtacaa atgatgacag cgaagtgacg
     5641 gcgcctagcc ggtcatcctt ttgacgattc cagtcccgag gataacctcc tctcgggatt
     5701 ggggggaatc tttggatcca gtagtcctcc ttgaactccg tccaacaggg cagattcaag
     5761 agtcataaga ctttcattaa tcatttcagt tgatcagaca tggtcgtgta gattctctta
     5821 atacgggaga tcttctagca gtttcagtga ccaacggtgc tttcattctc caggaactga
     5881 taccaaaggt tgtggacagg ccaaggggta cttcggatga ctctgtgctt gggcacagaa
     5941 agaggtcgta gtgtgccccc cgatagcgga ctcgacatga atcaactaag aaaggcaatc
     6001 tgcctcccat gatggacata agcaatagtt cacaaccatc ttgcatctca gtgaagtgta
     6061 cataactatt gagggctggg tcatctaagc atttcagtcg agaaaaaaac tgtagaccaa
     6121 aagaacaact agcaacactt ctcatccaga gacccatatc aagatgctag atccgggaga
     6181 ggtttatgat gaccctattg atccaattga gtcagaggct gaacccagag gaacccccac
     6241 tgtccccaac atcttgagga actccgacta caatctcaat tctcctttga tagaggatcc
     6301 tgccaaacta atgttagaat ggttgaagac agggaacaga ccttatcgga tgactttgac
     6361 agacaattgc tccaggtctt acaaagtttt gaaagattat ttcaagaaag tagatttggg
     6421 ttctctcaaa gtgggcggaa ctgctgcaca gtcaatggtt tctctctggt tgtgtggtgc
     6481 ccactctgaa tcaaacagga gccggagatg tataaccgac ttggcccatt tctattccaa
     6541 gtcatccccc atagagaagc tattgaattg tacgctagga aacagaggcc tgagaatccc
     6601 accagagggg gtgttaaatt gcctcgagag ggtcaattat gacaaggcat ttgggaggta
     6661 tctggccaac acgtattcct cttacttgtt tttccatgta atcaccttat acatgaatgc
     6721 cttagactgg gaagaggaaa agaccatcct ggcattatgg aaagatataa cctcagtgga
     6781 taccgagaag gacttggtca aattcaaaga tcaaatatgg ggactgttga ttgtgacaaa
     6841 ggactttgtt tactctcaga gttcaaactg tctttttgac agaaactaca cactgatgct
     6901 aaaggatctt ttcttgtctc gattcaactc cttaatgatt ctgctttctc cccctgagcc
     6961 ccgatactca gatgacttaa tatctcagct gtgccagcta tacatcgctg gggatcaagt
     7021 cttgtccatg tgtgggaact ccggctatga agtcatcaaa atattggagc catatgtcgt
     7081 gaacagtttg gtccagaggg cagagaagtt taggcctctc atccacccct tgggagactt
     7141 tcctatgttc ataaaagaca aggtgaatca acttgaaggg acttttggtc ccagtgcaaa
     7201 gaggtttttt agggttttag atcaattcga caacatacat gacttagtat ttgtgtatgg
     7261 ctgttacaga cattgggggc acccctatat agattatcgg aagggtctgt cgaaactata
     7321 tgatcaagtt cacattaaga aagtaataga taagtcctac caggagtgtt tagcaagtga
     7381 cttggccaga aggatcctca gatggggatt tgacaagtac tccaaatggt atctagattc
     7441 gagattcctt gcccgagacc accccttgac tccttatgtc aagacccaaa catggccacc
     7501 caaacatata gtagacttgg tgggggacac atggcataag ctcccgatca cgcagatctt
     7561 tgaaattcct gaatcaatgg acccgtcaga gatactggat gacaaatcac attctttcac
     7621 cagaacaagg ttagcttctt ggctgtcaga gaaccgaggg gggcctgttc ctagcgagaa
     7681 ggtcattatc acggccctgt ctaagccacc tgtcaatccc cgagagtttt tgaaatctat
     7741 agacctcgga ggattgccag atgaggattt gataattggc ctcaaaccaa aggaacggga
     7801 gttgaagatc gagggccgat tctttgctct aatgtcatgg aatctaagat tatattttgt
     7861 catcaccgaa aagctcttgg ccaactacat tttgccactt tttgacgcac tgactatgac
     7921 agacaacctg aacaaggtgt tcaaaaagtt gatcgacagg gtcaccgggc aagggctttt
     7981 ggactattct agggtcacat acgcatttca cctggactat gagaaatgga acaatcatca
     8041 aagattggag tcaacagagg atgtattctc tgtcctagat caggtgtttg gattgaagag
     8101 ggtgttttct agaacacacg agttttttca gaagtcctgg atctattatt cagacagatc
     8161 agacctcatt gggttatggg aggaccaaat atattgcttg gatatgtcta acggcccaac
     8221 ctgctggaat ggccaagatg gcgggctaga gggcttgcgg cagaagggct ggagtctagt
     8281 cagtttatta atgatagata gagaatctca aaccaggaac acaagaacca agatactagc
     8341 tcaaggagac aaccaggttc tgtgtcctac atacatgttg tcaccgggat tgtctcaaga
     8401 ggggcttctc tatgagttag agagcatatc aaggaatgca ctctcaatat accgagctat
     8461 cgaggaagga gcatctaagc tggggctgat catcaagaag gaagagacca tgtgtagtta
     8521 tgactttctc atatatggga agaccccctt atttcgaggc aacatattgg tgcctgaatc
     8581 caaaagatgg gcccgagtct cttgcatctc taacgaccaa atagtcaacc tcgccaatat
     8641 aatgtcgaca gtatccacca atgcgctgac agtggcacaa cactctcaat ctctgatcaa
     8701 acctatgagg gattttctgc tcatgtcagt acaggcagtt ttccactacc tgttgtttag
     8761 cccaatccta aaaggcagag tttataagat tctgagtgct gaaggggaga gctttctcct
     8821 agccatgtcg cggataatct acctagatcc ttctttggga ggggtgtctg gaatgtctct
     8881 cgggaggttc catatacgtc agttctcaga ccctgtctct gaagggttgt cattctggag
     8941 agagatctgg ttaggctctc atgagtcctg gattcacgcg ttgtgtcagg aggccgggaa
     9001 ccccgatctt ggagagagaa cactcgagag cttcactcgc cttttagaag atcctactac
     9061 cttaaatatc aaaggagggg ccagccctac cattctactc aaggatgcta tcagaaaggc
     9121 tctgtacgac gaggtggaca aggtggaaaa ttcagagttt cgagaggcaa tcctgctgtc
     9181 caagacccat agagataact ttatactctt tttaaaatct gttgagcctc tgttccctcg
     9241 atttctcagt gagctcttca gttcgtcttt cttgggaata ccggagtcaa tcattggact
     9301 gatacaaaac tcccggacaa taagaaggca gtttagaaag agtctctcaa gaactttaga
     9361 agagtccttc tacaactcag agatccacgg gattaatcgg ataacccaga cacctcaaag
     9421 ggtcggaagg gtgtggcctt gctcttcaga gagggcagat ctacttaggg agatctcttg
     9481 ggggaggaaa gtggtaggca cgacagttcc tcacccttcc gagatgttgg agttgtttcc
     9541 caaatcctcc atttcctgca cttgtggagc aacaggggga ggcaatccta gagtctctgt
     9601 atcagtactc ccgtccttcg atcagtcatt tttctcacgg ggccccctaa agggatactt
     9661 gggctcgtcc acctccatgt caacccagct attccatgca tgggaaaaag tcactaatgt
     9721 tcatgtggtg aaaagggctc tatcgttaaa agaatctata aactggttca tcaataggaa
     9781 ttccaatttg gctcaaactc taattggaaa catcatgtct ctgacaggcc ctgatttccc
     9841 tctagaagag gcccctgttt tcaaacggac agggtcagcc ttgcataggt tcaagtctgc
     9901 cagatacagc gaaggagggt attcttctgt ttgccctaac cttctctccc atatctctgt
     9961 tagtacagac actatgtctg atttgaccca aaacgggaag aactatgatt tcatgtttca
    10021 gccattgatg ctttatgcgc aaacatggac atcggaactg gtacagaggg atacaagact
    10081 tagagactcc acgtttcact ggcaccttcg gtgcaacaga tgtgtgaggc ccattgatga
    10141 tataacactg gaaacttctc agatcttcga gttcccggat gtgtcaaaaa ggatatccag
    10201 gatggtttct ggagctgtcc ctcaatttca gaagcttcct gatatccgtc taagaccagg
    10261 tgattttgaa tccctaagtg gtagagaaaa gtctcgccat atagggtcag ctcaggggct
    10321 cttatactca atcttagtag caattcacga ctcaggatac aatgatggga ccatcttccc
    10381 tgtcaacata tacggcaaag tttcccccag agactatttg agagggcttg caagagggat
    10441 cttgataggg tcctcgattt gcttcttgac acgaatgaca aatattaaca ttaacagacc
    10501 tcttgaattg atctcagggg taatttccta tattctcctg aggctggata atcatccctc
    10561 tctgtatata atgcttagag aaccgtctct tagaggagaa atattctcta tccctcagaa
    10621 aatccccgcc gcttacccaa ccactatgag agaaggcaac agatcgattt tgtgttacct
    10681 ccaacacgtg ctacgctatg agcgagaggc aatcacggcg tccccggaga atgactggct
    10741 gtggatcttc tcagacttca gaagtgtgaa aatgacgtac ttgaccctca ttacctacca
    10801 gtctcacctc ctactccaga gggttgagag aaacttgtct aagagtatga gagctactct
    10861 gcgacaaatg agttccttaa tgaggcaagt gctgggtggg cacggagaag ataccttgga
    10921 gtcagacgat gacattcaac gattactaaa agactctttg cgaaagacaa ggtgggtgga
    10981 tcaagaggtg cgccatgcag ctagaaccat gaatggagat tacagccccg acaagaaagt
    11041 atcccacaag gcaggatgtt cagaatgggt ctgctctgct caacagattg ccgtctccac
    11101 ctcagccaac ccggcccctg tctcagagct tgacattagg gccctctcta agaggtttca
    11161 aaaccccttg atctcgggcc tgagagtggt tcagtgggca accggtgccc attataagct
    11221 taagcctatt ctagatgatc taaatgtttt cccatctctc tgtcttgtaa ttggagacgg
    11281 gtcaggggga atatcaaggg cagttctcaa catgtttcca gattctaagc ttgtgttcaa
    11341 cagcctattg gaggtgaatg atctgatggc ttccggaaca catccactgc ctccttcagc
    11401 aatcatgagt ggaggagatg acatcatctc cagagtgata gactttgact caatatggga
    11461 gaaaccgtcc gacctgagga actcggccac ctggagatac ttccagtcgg ttcaaaaaca
    11521 ggtcaacatg tcgtatgacc tcattatttg tgatgcagaa gttactgata ttgcatctat
    11581 caaccggata actctgttga tgtctgattt cgcattgtct atagatggac cactttatct
    11641 ggtcttcaaa acttacggga ctatgctagt aaacccggac tataaagcta ttcaacatct
    11701 gtcaagagcg ttcccttcgg tcacagggtt tgtaacccaa gtaacttcat ccttttcttc
    11761 tgagctatac ctccggttct ctaaacgagg aaagtttttc agggacgccg agtacttgac
    11821 ctcttccacc cttcgagaga tgagccttgt gttgttcaat tgcagcagcc ccaaaagtga
    11881 gatgcagaga gctcgttcct taaactatca agacctggta aggggatttc ctgaagagat
    11941 catatcaaat ccttacaacg aaatgatcat aactctgatt gacaatgatg tagagtcctt
    12001 cctagtccac aagatggtgg atgatcttga gctacagagg ggaactctgt ctaaagtggc
    12061 tatcattata tccatcatga tcgttttttc caatagagtc ttcaacattt ccaaaccttt
    12121 gactgacccc ttgttctacc ccccatctga tcctaaaatc ctgaggcact tcaacatatg
    12181 ttgcagtact atgatgtatc tatctaccgc tttaggcgac gtccctaact tcgcaagact
    12241 tcatgacctg tataacagac ctataacttg ttacttcaga aaacaagtta ttcgagggaa
    12301 tatttatcta tcttggagtt ggtccgatga taccccagtg ttcaagagag tagcctgtaa
    12361 ttctagcttg agtctgtcat ctcactggat caggttgatc tacaagatag tgaagactac
    12421 cagactcatt ggcagcataa aagacctatc aggagaggta gaacgacatc ttcatgggta
    12481 taacagatgg atcaccctcg aggatatccg atctagatca tccctactag attacagttg
    12541 tttgtaagcc ggatattacc gaaagcctgt gcatgctaaa attcttgtat gatgcatctt
    12601 gaaaaaaaca agatcttgaa tccggacctc tggttgtttg attgtttttt ccatctttat
    12661 tgtttttttg ttaagcgtgg gtcggcatgg catctccacc tcctcgcggt ccgacctggg
    12721 catccgaagg aggacgcacg tccactcgga tggctaaggg agggcgggga tccggctgct
    12781 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
    12841 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaagtcgc gcttggcgta
    12901 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
    12961 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
    13021 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
    13081 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
    13141 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
    13201 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
    13261 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
    13321 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
    13381 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
    13441 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
    13501 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
    13561 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
    13621 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
    13681 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
    13741 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    13801 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    13861 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    13921 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    13981 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    14041 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    14101 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    14161 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccaccctt
    14221 ggaggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    14281 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    14341 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    14401 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    14461 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    14521 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    14581 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    14641 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    14701 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    14761 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    14821 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    14881 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    14941 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    15001 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    15061 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
    15121 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
    15181 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
    15241 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
    15301 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
    15361 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
    15421 agggattttg ccgatttcgg cctattggtt aaaaaaatga gctgatttaa caaaaattta
    15481 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa
    15541 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
    15601 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
    15661 aacgacggcc agtgagcgcg cctagttatt aatagtaatc aattacgggg tcattagttc
    15721 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
    15781 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
    15841 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
    15901 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
    15961 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
    16021 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
    16081 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
    16141 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
    16201 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
    16261 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
    16321 aagctggcta gattaagcgt ctgatgagtc cgtgaggacg aaacccggcg taccgggtc
//
