LOCUS       sequence-343689-       16265 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 16265)
  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 16265)
  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.mCherry-GCaMP7s
FEATURES             Location/Qualifiers
     source          1..16265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     253..272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(491..595)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      622..1078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(709..728)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     919..940
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1204..1226
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1218..1235
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1219..1235
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        1305..1608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1609..1812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1763..1783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1857..1876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1857..1875
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5260..5269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5266..5973
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5411..5429)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5679..5698
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5989..6009
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6013..6033
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6037..6057
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      6155..6164
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6170..6187
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6191..6223
                     /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(6227..6235,6236..6250)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      6257..6266
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6278..6334
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     6547..6568
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6626..6647)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6887..6906)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             7022..7465
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             9635..9646
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(11438..11449)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(14645..14663)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      14659..14706
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(14734..14750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14734..14750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14747..14769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14758..14774
                     /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(14782..14788,14789..14806,14807..14812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14827..14848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14965..14982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15136..15724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15216..15235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15895..16265,1..421,422..490))
                     /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"
ORIGIN
        1 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
       61 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
      121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
      181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
      241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
      301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
      361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
      421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
      481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      601 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
      661 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
      721 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
      781 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
      841 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
      901 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      961 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaaat
     1021 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     1081 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1141 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     1201 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcctagtta ttaatagtaa
     1261 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     1321 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1381 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1441 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1501 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1561 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1621 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1681 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1741 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1801 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
     1861 acgactcact atagggagac ccaagctggc tagattaagc gtctgatgag tccgtgagga
     1921 cgaaacccgg cgtaccgggt cacgcttaac aacaaaacca gagaagaaaa agacagcgtc
     1981 aattgcaaag caaaaatgta acacccctac aatggatgcc gacaagattg tgttcaaagt
     2041 caataatcag gtggtctctt tgaagcctga gattatcgtg gatcaatatg agtacaagta
     2101 ccctgccatc aaggatttga aaaagccttg tatcacccta gggaaagccc ccgacttgaa
     2161 caaagcatac aaatcagttt tatcaggcat gaatgccgcc aaacttgatc cggatgatgt
     2221 atgctcctac ttggcagcag caatgcagtt ctttgagggg acatgtccgg aagactggac
     2281 cagctatgga atcctgattg cacgaaaagg agataggatc accccaaact ctctagtgga
     2341 gataaagcgt actgatgtag aagggaattg ggctctgaca ggaggcatgg aattgacaag
     2401 ggaccccact gtctctgaac atgcatcttt agtcggtctt ctcctgagtc tgtacaggtt
     2461 gagcaaaata tcaggacaga acactggtaa ctataagaca aacattgcag ataggataga
     2521 gcagattttc gagacagcac cttttgttaa gatcgtggaa caccataccc taatgacaac
     2581 tcacaagatg tgtgctaatt ggagtactat accgaacttc agatttttgg ccggaaccta
     2641 cgacatgttt ttctcacgga ttgagcatct atattcggca atcagagtgg gcacagtcgt
     2701 caccgcttat gaagactgct caggactggt atcgtttaca gggttcataa agcagatcaa
     2761 tctcaccgca agggaagcaa tactatattt cttccacaag aactttgagg aagagataag
     2821 aagaatgttc gagccagggc aagagacagc tgttcctcac tcttatttca tccacttccg
     2881 ttcactaggc ttgagtggga agtctcctta ttcatcgaat gctgtcggtc atgtgttcaa
     2941 tctcattcac tttgttggat gctacatggg tcaagtcaga tctctaaatg cgacggttat
     3001 tgctgcatgt gcccctcatg agatgtctgt tctagggggc tatttgggag aggaattctt
     3061 cggaaaaggg acatttgaaa gaaggttctt cagagacgag aaagaacttc aagaatatga
     3121 ggcggctgaa ctaacaaagt ccgacgtggc actggcagat gacggaaccg tcaactctga
     3181 tgacgaggac tatttctctg gtgaaaccag aagtccagaa gctgtctata ctcgaatcat
     3241 gatgaatgga ggtcgactga agagatctca tatacggaga tatgtctcag tcagttccaa
     3301 tcatcaagcc cgtccaaact cattcgccga atttttaaac aagacgtatt cgaatgactc
     3361 ataaggagtt gattgacagg gtgccagaaa tctatagatt gtatatatcc atcatgaaaa
     3421 aaactaacac tcctcctttc aaaccatccc aaatatgagc aagatctttg ttaatccgag
     3481 tgcaatcaga gccggtctgg ccgatcttga gatggccgaa gagactgttg atctgatcaa
     3541 cagaaacata gaagacaatc aggctcatct ccagggagaa cccatagaag tggacaacct
     3601 ccctgaggac atgaagcgac ttcacctgga cgatgaaaaa tcgtccaacc ttggtgagat
     3661 ggttagggtg ggagaaggca agtatcgaga ggactttcag atggatgagg gagaggaccc
     3721 caacctcctg ttccaatcgt acctggataa tgttggagtc caaatagtca gacaaatgag
     3781 gtcaggagag agattcctca agatatggtc acagaccgta gaggaaattg tatcctatgt
     3841 cacggtcaac tttcctaacc ctccaagaag gtcttcggag gataaatcaa cccagactac
     3901 tggcagagag ctcaagaagg agacaacgtc tgctttctct cagagagaaa gccaaccttc
     3961 gaaagctagg atggtggctc aagttgcccc tggtcctcca gcccttgaat ggtcagccac
     4021 caatgaagaa gatgatctat cagtagaggc tgagatcgct catcagattg ctgaaagctt
     4081 ttccaagaag tacaagtttc cctcccgatc ttcaggaata ttcttgtata attttgagca
     4141 actgaagatg aaccttgatg acatagttaa agaggcaaaa aatgtaccgg gcgtgacccg
     4201 tctggcccat gatggatcca aaatccccct gagatgcgta ctgggatggg tcgctttggc
     4261 taattccaaa aaattccaat tactagtcga ggctgacaag ctaagcaaaa tcatgcaaga
     4321 tgatttgaat cgctatacat cctgctaacc gagttttcga actcagtccc tccagataat
     4381 gaaaactgag atgttatgga gtagacatga aaaaaacagg caacaccact gataaaatga
     4441 acgttctacg caagatagtg aaaaaatgta gggatgagga cactcaaaag ccctctcctg
     4501 tgtcagcccc tccgtatgac gatgacctgt ggcttccacc tcctgaatat gtcccgctga
     4561 aagaactcac aagcaagaag aacatgagga acttttgtgt caacggggag gttaaagcgt
     4621 gtagcccaaa tggttactca ttcaggattt tgcggcacat tctgagatca ttcaacgaga
     4681 tatactctgg gaatcatagg atgattgggt tagtcaaaat tgttgttgga ctagctttat
     4741 caggagctcc agtacctgag ggcatgaact gggtatacaa attgaggaga acccttatat
     4801 tccagtgggc tgattccagg ggccctcttg aaggggagga gttagattac tctcaagaga
     4861 tcacttggga tgatgatact gaattcgtcg gattgcaaat aagagtgagc gcaagacaat
     4921 gtcatattca aggcaggatc tggtgtatca acacgaactc gagggcatgt caactatggt
     4981 ctgacatgtc tcttcagaca caaaggtctg aagaggacaa agactcttct ctgcttctag
     5041 aataatcaga ttatatcctg caagtgtatc acttgtttac ctctggagga gagagcatac
     5101 aggcttgact ccgatccttg ggagcaatag aacaaaaaaa cacacgttat ggtgccgtta
     5161 aatcgctgca ttttatcaaa gtcaagttga taacctttac attttgagcc tcttggatgt
     5221 gaaaaaaact attaacatcc ctcaaaagac cccggtaaag ccaccatggt gagcaagggc
     5281 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     5341 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     5401 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     5461 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     5521 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     5581 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     5641 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     5701 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     5761 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     5821 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     5881 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     5941 cactccaccg gcggcatgga cgagctgtac aagagatctc gagctgatcc aaaaaagaag
     6001 agaaaggtag atccaaaaaa gaagagaaag gtagatccaa aaaagaagag aaaggtagga
     6061 tccagataac tgatcataat cagccatacc acatttggaa ttcgtcgagg gggcgcgccg
     6121 ctagacatga aaaaaactaa cactcctccg gtacgccacc atgggttctc atcatcatca
     6181 tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc tgtacgacga
     6241 tgacgataag gatctcgcca ccatggtcga ctcatcacgt cgtaagtgga ataagacagg
     6301 tcacgcagtc agagctatag gtcggctgag ctcactcgag aacgtctata tcaaggccga
     6361 caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcgg
     6421 cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     6481 gcccgacaac cactacctga gcgtgcagtc caaactttcg aaagacccca acgagaagcg
     6541 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggagga
     6601 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     6661 gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga ccctgaagtt
     6721 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     6781 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     6841 cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg acggcaacta
     6901 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagcttaa
     6961 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacctgcc
     7021 ggaccaactg actgaagagc agatcgcaga atttaaagag ctgttctccc tatttgacaa
     7081 ggacggggat gggacaataa caaccaagga gctggggacg gtgatgcggt ctctggggca
     7141 gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg
     7201 cacaatcgac ttccctgagt tcctgacaat gatggcaaga aaaatgaaat acacagacag
     7261 tgaagaagaa attagagaag cgttccgtgt gtttgataag gatggcaatg gctacatcag
     7321 tgcagcagag cttcgccacg tgatgacaaa ccttggagag aagttaacag atgaagaggt
     7381 tgatgaaatg atcagggaag cagacatcga tggggatggt caggtaaact acgaagagtt
     7441 tgtacaaatg atgacagcga agtgacggcg cctagccggt catccttttg acgattccag
     7501 tcccgaggat aacctcctct cgggattggg gggaatcttt ggatccagta gtcctccttg
     7561 aactccgtcc aacagggcag attcaagagt cataagactt tcattaatca tttcagttga
     7621 tcagacatgg tcgtgtagat tctcttaata cgggagatct tctagcagtt tcagtgacca
     7681 acggtgcttt cattctccag gaactgatac caaaggttgt ggacaggcca aggggtactt
     7741 cggatgactc tgtgcttggg cacagaaaga ggtcgtagtg tgccccccga tagcggactc
     7801 gacatgaatc aactaagaaa ggcaatctgc ctcccatgat ggacataagc aatagttcac
     7861 aaccatcttg catctcagtg aagtgtacat aactattgag ggctgggtca tctaagcatt
     7921 tcagtcgaga aaaaaactgt agaccaaaag aacaactagc aacacttctc atccagagac
     7981 ccatatcaag atgctagatc cgggagaggt ttatgatgac cctattgatc caattgagtc
     8041 agaggctgaa cccagaggaa cccccactgt ccccaacatc ttgaggaact ccgactacaa
     8101 tctcaattct cctttgatag aggatcctgc caaactaatg ttagaatggt tgaagacagg
     8161 gaacagacct tatcggatga ctttgacaga caattgctcc aggtcttaca aagttttgaa
     8221 agattatttc aagaaagtag atttgggttc tctcaaagtg ggcggaactg ctgcacagtc
     8281 aatggtttct ctctggttgt gtggtgccca ctctgaatca aacaggagcc ggagatgtat
     8341 aaccgacttg gcccatttct attccaagtc atcccccata gagaagctat tgaattgtac
     8401 gctaggaaac agaggcctga gaatcccacc agagggggtg ttaaattgcc tcgagagggt
     8461 caattatgac aaggcatttg ggaggtatct ggccaacacg tattcctctt acttgttttt
     8521 ccatgtaatc accttataca tgaatgcctt agactgggaa gaggaaaaga ccatcctggc
     8581 attatggaaa gatataacct cagtggatac cgagaaggac ttggtcaaat tcaaagatca
     8641 aatatgggga ctgttgattg tgacaaagga ctttgtttac tctcagagtt caaactgtct
     8701 ttttgacaga aactacacac tgatgctaaa ggatcttttc ttgtctcgat tcaactcctt
     8761 aatgattctg ctttctcccc ctgagccccg atactcagat gacttaatat ctcagctgtg
     8821 ccagctatac atcgctgggg atcaagtctt gtccatgtgt gggaactccg gctatgaagt
     8881 catcaaaata ttggagccat atgtcgtgaa cagtttggtc cagagggcag agaagtttag
     8941 gcctctcatc caccccttgg gagactttcc tatgttcata aaagacaagg tgaatcaact
     9001 tgaagggact tttggtccca gtgcaaagag gttttttagg gttttagatc aattcgacaa
     9061 catacatgac ttagtatttg tgtatggctg ttacagacat tgggggcacc cctatataga
     9121 ttatcggaag ggtctgtcga aactatatga tcaagttcac attaagaaag taatagataa
     9181 gtcctaccag gagtgtttag caagtgactt ggccagaagg atcctcagat ggggatttga
     9241 caagtactcc aaatggtatc tagattcgag attccttgcc cgagaccacc ccttgactcc
     9301 ttatgtcaag acccaaacat ggccacccaa acatatagta gacttggtgg gggacacatg
     9361 gcataagctc ccgatcacgc agatctttga aattcctgaa tcaatggacc cgtcagagat
     9421 actggatgac aaatcacatt ctttcaccag aacaaggtta gcttcttggc tgtcagagaa
     9481 ccgagggggg cctgttccta gcgagaaggt cattatcacg gccctgtcta agccacctgt
     9541 caatccccga gagtttttga aatctataga cctcggagga ttgccagatg aggatttgat
     9601 aattggcctc aaaccaaagg aacgggagtt gaagatcgag ggccgattct ttgctctaat
     9661 gtcatggaat ctaagattat attttgtcat caccgaaaag ctcttggcca actacatttt
     9721 gccacttttt gacgcactga ctatgacaga caacctgaac aaggtgttca aaaagttgat
     9781 cgacagggtc accgggcaag ggcttttgga ctattctagg gtcacatacg catttcacct
     9841 ggactatgag aaatggaaca atcatcaaag attggagtca acagaggatg tattctctgt
     9901 cctagatcag gtgtttggat tgaagagggt gttttctaga acacacgagt tttttcagaa
     9961 gtcctggatc tattattcag acagatcaga cctcattggg ttatgggagg accaaatata
    10021 ttgcttggat atgtctaacg gcccaacctg ctggaatggc caagatggcg ggctagaggg
    10081 cttgcggcag aagggctgga gtctagtcag tttattaatg atagatagag aatctcaaac
    10141 caggaacaca agaaccaaga tactagctca aggagacaac caggttctgt gtcctacata
    10201 catgttgtca ccgggattgt ctcaagaggg gcttctctat gagttagaga gcatatcaag
    10261 gaatgcactc tcaatatacc gagctatcga ggaaggagca tctaagctgg ggctgatcat
    10321 caagaaggaa gagaccatgt gtagttatga ctttctcata tatgggaaga cccccttatt
    10381 tcgaggcaac atattggtgc ctgaatccaa aagatgggcc cgagtctctt gcatctctaa
    10441 cgaccaaata gtcaacctcg ccaatataat gtcgacagta tccaccaatg cgctgacagt
    10501 ggcacaacac tctcaatctc tgatcaaacc tatgagggat tttctgctca tgtcagtaca
    10561 ggcagttttc cactacctgt tgtttagccc aatcctaaaa ggcagagttt ataagattct
    10621 gagtgctgaa ggggagagct ttctcctagc catgtcgcgg ataatctacc tagatccttc
    10681 tttgggaggg gtgtctggaa tgtctctcgg gaggttccat atacgtcagt tctcagaccc
    10741 tgtctctgaa gggttgtcat tctggagaga gatctggtta ggctctcatg agtcctggat
    10801 tcacgcgttg tgtcaggagg ccgggaaccc cgatcttgga gagagaacac tcgagagctt
    10861 cactcgcctt ttagaagatc ctactacctt aaatatcaaa ggaggggcca gccctaccat
    10921 tctactcaag gatgctatca gaaaggctct gtacgacgag gtggacaagg tggaaaattc
    10981 agagtttcga gaggcaatcc tgctgtccaa gacccataga gataacttta tactcttttt
    11041 aaaatctgtt gagcctctgt tccctcgatt tctcagtgag ctcttcagtt cgtctttctt
    11101 gggaataccg gagtcaatca ttggactgat acaaaactcc cggacaataa gaaggcagtt
    11161 tagaaagagt ctctcaagaa ctttagaaga gtccttctac aactcagaga tccacgggat
    11221 taatcggata acccagacac ctcaaagggt cggaagggtg tggccttgct cttcagagag
    11281 ggcagatcta cttagggaga tctcttgggg gaggaaagtg gtaggcacga cagttcctca
    11341 cccttccgag atgttggagt tgtttcccaa atcctccatt tcctgcactt gtggagcaac
    11401 agggggaggc aatcctagag tctctgtatc agtactcccg tccttcgatc agtcattttt
    11461 ctcacggggc cccctaaagg gatacttggg ctcgtccacc tccatgtcaa cccagctatt
    11521 ccatgcatgg gaaaaagtca ctaatgttca tgtggtgaaa agggctctat cgttaaaaga
    11581 atctataaac tggttcatca ataggaattc caatttggct caaactctaa ttggaaacat
    11641 catgtctctg acaggccctg atttccctct agaagaggcc cctgttttca aacggacagg
    11701 gtcagccttg cataggttca agtctgccag atacagcgaa ggagggtatt cttctgtttg
    11761 ccctaacctt ctctcccata tctctgttag tacagacact atgtctgatt tgacccaaaa
    11821 cgggaagaac tatgatttca tgtttcagcc attgatgctt tatgcgcaaa catggacatc
    11881 ggaactggta cagagggata caagacttag agactccacg tttcactggc accttcggtg
    11941 caacagatgt gtgaggccca ttgatgatat aacactggaa acttctcaga tcttcgagtt
    12001 cccggatgtg tcaaaaagga tatccaggat ggtttctgga gctgtccctc aatttcagaa
    12061 gcttcctgat atccgtctaa gaccaggtga ttttgaatcc ctaagtggta gagaaaagtc
    12121 tcgccatata gggtcagctc aggggctctt atactcaatc ttagtagcaa ttcacgactc
    12181 aggatacaat gatgggacca tcttccctgt caacatatac ggcaaagttt cccccagaga
    12241 ctatttgaga gggcttgcaa gagggatctt gatagggtcc tcgatttgct tcttgacacg
    12301 aatgacaaat attaacatta acagacctct tgaattgatc tcaggggtaa tttcctatat
    12361 tctcctgagg ctggataatc atccctctct gtatataatg cttagagaac cgtctcttag
    12421 aggagaaata ttctctatcc ctcagaaaat ccccgccgct tacccaacca ctatgagaga
    12481 aggcaacaga tcgattttgt gttacctcca acacgtgcta cgctatgagc gagaggcaat
    12541 cacggcgtcc ccggagaatg actggctgtg gatcttctca gacttcagaa gtgtgaaaat
    12601 gacgtacttg accctcatta cctaccagtc tcacctccta ctccagaggg ttgagagaaa
    12661 cttgtctaag agtatgagag ctactctgcg acaaatgagt tccttaatga ggcaagtgct
    12721 gggtgggcac ggagaagata ccttggagtc agacgatgac attcaacgat tactaaaaga
    12781 ctctttgcga aagacaaggt gggtggatca agaggtgcgc catgcagcta gaaccatgaa
    12841 tggagattac agccccgaca agaaagtatc ccacaaggca ggatgttcag aatgggtctg
    12901 ctctgctcaa cagattgccg tctccacctc agccaacccg gcccctgtct cagagcttga
    12961 cattagggcc ctctctaaga ggtttcaaaa ccccttgatc tcgggcctga gagtggttca
    13021 gtgggcaacc ggtgcccatt ataagcttaa gcctattcta gatgatctaa atgttttccc
    13081 atctctctgt cttgtaattg gagacgggtc agggggaata tcaagggcag ttctcaacat
    13141 gtttccagat tctaagcttg tgttcaacag cctattggag gtgaatgatc tgatggcttc
    13201 cggaacacat ccactgcctc cttcagcaat catgagtgga ggagatgaca tcatctccag
    13261 agtgatagac tttgactcaa tatgggagaa accgtccgac ctgaggaact cggccacctg
    13321 gagatacttc cagtcggttc aaaaacaggt caacatgtcg tatgacctca ttatttgtga
    13381 tgcagaagtt actgatattg catctatcaa ccggataact ctgttgatgt ctgatttcgc
    13441 attgtctata gatggaccac tttatctggt cttcaaaact tacgggacta tgctagtaaa
    13501 cccggactat aaagctattc aacatctgtc aagagcgttc ccttcggtca cagggtttgt
    13561 aacccaagta acttcatcct tttcttctga gctatacctc cggttctcta aacgaggaaa
    13621 gtttttcagg gacgccgagt acttgacctc ttccaccctt cgagagatga gccttgtgtt
    13681 gttcaattgc agcagcccca aaagtgagat gcagagagct cgttccttaa actatcaaga
    13741 cctggtaagg ggatttcctg aagagatcat atcaaatcct tacaacgaaa tgatcataac
    13801 tctgattgac aatgatgtag agtccttcct agtccacaag atggtggatg atcttgagct
    13861 acagagggga actctgtcta aagtggctat cattatatcc atcatgatcg ttttttccaa
    13921 tagagtcttc aacatttcca aacctttgac tgaccccttg ttctaccccc catctgatcc
    13981 taaaatcctg aggcacttca acatatgttg cagtactatg atgtatctat ctaccgcttt
    14041 aggcgacgtc cctaacttcg caagacttca tgacctgtat aacagaccta taacttgtta
    14101 cttcagaaaa caagttattc gagggaatat ttatctatct tggagttggt ccgatgatac
    14161 cccagtgttc aagagagtag cctgtaattc tagcttgagt ctgtcatctc actggatcag
    14221 gttgatctac aagatagtga agactaccag actcattggc agcataaaag acctatcagg
    14281 agaggtagaa cgacatcttc atgggtataa cagatggatc accctcgagg atatccgatc
    14341 tagatcatcc ctactagatt acagttgttt gtaagccgga tattaccgaa agcctgtgca
    14401 tgctaaaatt cttgtatgat gcatcttgaa aaaaacaaga tcttgaatcc ggacctctgg
    14461 ttgtttgatt gttttttcca tctttattgt ttttttgtta agcgtgggtc ggcatggcat
    14521 ctccacctcc tcgcggtccg acctgggcat ccgaaggagg acgcacgtcc actcggatgg
    14581 ctaagggagg gcggggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
    14641 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
    14701 tttttgctga aagtcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    14761 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    14821 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    14881 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    14941 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    15001 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    15061 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    15121 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    15181 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    15241 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    15301 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    15361 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    15421 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    15481 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    15541 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
    15601 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    15661 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    15721 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    15781 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
    15841 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
    15901 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
    15961 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
    16021 tgcaatgata ccgcgagacc cacccttgga ggctccagat ttatcagcaa taaaccagcc
    16081 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
    16141 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
    16201 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
    16261 cggtt
//
