LOCUS       sequence-79121-e       12590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coexpression plasmid of QuasAr1 and CheRiff for all-optical 
            electrophysiology.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12590)
  AUTHORS   Hochbaum DR, Zhao Y, Farhi SL, Klapoetke N, Werley CA, Kapoor V, Zou
            P, Kralj JM, Maclaurin D, Smedemark-Margulies N, Saulnier JL, 
            Boulting GL, Straub C, Cho YK, Melkonian M, Wong GK, Harrison DJ, 
            Murthy VN, Sabatini BL, Boyden ES, Campbell RE, Cohen AE
  TITLE     All-optical electrophysiology in mammalian neurons using engineered 
            microbial rhodopsins.
  JOURNAL   Nat Methods. 2014 Aug;11(8):825-33. doi: 10.1038/nmeth.3000. Epub 
            2014 Jun 22.
  PUBMED    24952910
REFERENCE   2  (bases 1 to 12590)
  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"; doi: 
            "10.1038/nmeth.3000"; journalName: "Nat Methods"; date: "2014-08"; 
            volume: "11"; issue: "8"; pages: "825-33"
COMMENT     Sequence Label: DRH335: FCK-Optopatch1
FEATURES             Location/Qualifiers
     source          1..12590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2602..3891
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             3904..4662
                     /codon_start=1
                     /product="achaerhodopsin-3 (Arch) derivative with improved 
                     voltage sensitivity (Hochbaum et al., 2014)"
                     /label=QuasAr2
                     /translation="MVSIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT
                     DKDAREYYAVTILVSGIASAAYLSMFFGIGLTEVSVGGEMLDIYYARYAHWLFTTPLLL
                     LHLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYVLA
                     TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD
                     VTAKVGFGFILLRSRAILGDTEAPEPSAGAD"
     CDS             4669..5376
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEYNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             5470..5526
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5527..6471
                     /codon_start=1
                     /product="blue light-activated optogenetic actuator derived
                     from Scherffelia dubia channelrhodopsin (Hochbaum et al., 
                     2014)"
                     /label=CheRiff
                     /translation="GGAPAPDAHSAPPGNDSAGGSEYHAPAGYQVNPPYHPVHGYEEQC
                     SSIYIYYGALWEQETARGFQWFAVFLSALFLAFYGWHAYKASVGWEEVYVCSVELIKVI
                     LEIYFEFTSPAMLFLYGGNITPWLRYAEWLLTCPVILIHLSNITGLSEAYNKRTMALLV
                     SDLGTICMGVTAALATGWVKWLFYCIGLVYGTQTFYNAGIIYVESYYIMPAGGCKKLVL
                     AMTAVYYSSWLMFPGLFIFGPEGMHTLSVAGSTIGHTIADLLSKNIWGLLGHFLRIKIH
                     EHIIMYGDIRRPVSSQFLGRKVDVLAFVTEEDKV"
     CDS             join(6541..6543,6544..7257)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6541..6543,6544..7254)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6583..6604)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6844..6863)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7191..7212
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7282..7870
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7335..7355)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7753..7764)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7873..7889)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7874..7890)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8395..8575
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8601..8618)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8607..8831
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8877..9305
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8964..8983)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9174..9195
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9314..9334)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9319..9648
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9499..9634
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9561..9580
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9696..9743
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9762..10136
                     /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    10266..10387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10303..10322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10357..10376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10436..10452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10436..10452)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10449..10471)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10460..10476
                     /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(10484..10490,10491..10508,10509..10514))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10529..10550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10667..10684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10838..11426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10918..10937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11597..12388,12389..12457))
                     /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     12220..12239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12458..12562)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta acattatggc cttaggtcac ttcatctcca tggggttctt
     2641 cttctgattt tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag
     2701 ggtcacatca gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat
     2761 gccttggggg catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca
     2821 atgagtttag gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg
     2881 gtgagtccag ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga
     2941 agcacaagca gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg
     3001 aggatgggtg gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga
     3061 cagatagggg tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac
     3121 ttcttctcac gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg
     3181 gaaagaaaca aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa
     3241 agaaaccatt acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt
     3301 aaaccttaga acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga
     3361 tgaagttgcc agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc
     3421 cactagccgt taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa
     3481 aagggcccta tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag
     3541 tttgttcaca tccccttctc caaccccctc agtacatcac cctgggagaa caaggtccac
     3601 ttgcttctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg caacggagga
     3661 gcaggttttg aagtgaaagg caggcaggtg ttggggaggc agttaccggg gcaacgggaa
     3721 cagggcgttt cggaggtggt tgccatgggg acctggatgc tgacgaaggc tcgcgaggct
     3781 gtgagcagcc acagtgccct gctcagaagc cccaagctcg tcaatcaagc tggttctcca
     3841 tttgcactca ggagcacggg caggcgagtg gcccctagtt ctgggggcag cgggggatcc
     3901 accatggtaa gtatcgctct gcaggctggt tacgacctac tgggtgacgg cagacctgaa
     3961 accctgtggc tgggcatcgg cactctgctg atgctgattg gaaccttcta ctttctggtc
     4021 cgcggatggg gagtcaccga taaggatgcc cgggaatatt acgctgtgac tatcctggtg
     4081 tccggaatcg catccgccgc atatctgtct atgttctttg gtatcgggct tactgaggtg
     4141 tccgtcgggg gcgaaatgtt ggatatctat tatgccaggt acgcccattg gctgtttacc
     4201 accccacttc tgctgctgca cctggccctt ctcgctaagg tggatcgggt gaccatcggc
     4261 accctggtgg gtgtggacgc cctgatgatc gtcactggcc tcatcggagc cttgagccac
     4321 acggccatag ccagatacag ttggtggttg ttctctacaa tttgcatgat agtggtgctc
     4381 tatgttctgg ctacatccct gcgatctgct gcaaaggagc ggggccccga ggtggcatct
     4441 acctttaaca ccctgacagc tctggtcttg gtgctgtgga ccgcttaccc tatcctgtgg
     4501 atcataggca ctgagggcgc tggcgtggtg ggcctgggca tcgaaactct gctgtttatg
     4561 gtgttggacg tgactgccaa ggtcggcttt ggctttatcc tgttgagatc ccgggctatt
     4621 ctgggcgaca ccgaggcacc agaacccagt gccggtgccg atacgttgat ggtgagcaag
     4681 ggcgaggagt ataacatggc catcatcaag gagttcatgc gcttcaaggt gcgcatggag
     4741 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     4801 ggctttcaga ccgctaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     4861 atcctgtccc ctcatttcac ctacggctcc aaggcctacg tgaagcaccc cgccgacatc
     4921 cccgactact tcaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaactac
     4981 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     5041 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtgat gcagaagaag
     5101 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggtgc cctgaagggc
     5161 aagatcaaga tgaggctgaa gctgaaggac ggcggccact acacctccga ggtcaagacc
     5221 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acatcgtcga catcaagttg
     5281 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     5341 cgccactcca ccggcggcat ggacgagctg tacaagagta gaatcacaag cgaaggcgag
     5401 tacatccccc tggatcaaat agacataaat gtaggtggat tttgttatga gaatgaagtg
     5461 ggctccggag ccacgaactt ctctctgtta aagcaagcag gagacgtgga agaaaacccc
     5521 ggtcccggcg gagctcctgc tccagacgct cacagcgccc cacctggaaa cgattctgcc
     5581 ggaggcagtg agtaccatgc cccagctgga tatcaagtga atccacccta ccaccccgtg
     5641 catgggtatg aggaacagtg cagctccatc tacatctact atggggccct gtgggagcag
     5701 gaaacagcta ggggcttcca gtggtttgcc gtgttcctgt ctgccctgtt tctggctttc
     5761 tacggctggc acgcctataa ggccagcgtg ggatgggagg aagtgtacgt gtgctccgtg
     5821 gagctgatca aagtgattct ggagatctat ttcgagttca ccagtcctgc tatgctgttc
     5881 ctgtacggag ggaacattac cccatggctg agatatgccg aatggctgct gacatgtccc
     5941 gtgatcctga ttcatctgtc taacatcacc ggcctgagtg aggcatacaa taagcggaca
     6001 atggctctgc tggtgtccga cctgggaact atttgcatgg gagtgacagc cgctctggcc
     6061 actgggtggg tgaagtggct gttttactgt atcggcctgg tgtatggaac ccagacattc
     6121 tacaacgctg gaatcatcta cgtggagtct tactatatca tgcctgccgg cggctgtaag
     6181 aaactggtgc tggccatgac tgccgtgtac tattctagtt ggctgatgtt tcccggcctg
     6241 ttcatctttg ggcctgaagg catgcacacc ctgagcgtgg ctgggtccac tattggccat
     6301 accatcgccg acctgctgtc caagaatatt tggggactgc tggggcactt cctgcggatc
     6361 aaaattcacg agcatatcat tatgtacggc gatatcagga gaccagtgag ctcccagttt
     6421 ctgggacgca aggtggacgt gctggccttc gtgacagagg aagataaagt ggcggccgcc
     6481 aagagcagga tcaccagcga gggcgagtac atccccctgg accagatcga catcaacgtg
     6541 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     6601 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     6661 aagctgaccc tgaagttcat ttgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     6721 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     6781 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     6841 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     6901 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     6961 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     7021 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     7081 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     7141 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     7201 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagaa
     7261 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     7321 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     7381 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     7441 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     7501 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     7561 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     7621 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     7681 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     7741 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     7801 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     7861 gcctccccgc atcgataccg tcgacctcga gacctagaaa aacatggagc aatcacaagt
     7921 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     7981 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     8041 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     8101 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag
     8161 aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta
     8221 gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta
     8281 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt
     8341 gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct
     8401 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     8461 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     8521 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc
     8581 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     8641 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     8701 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     8761 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     8821 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     8881 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     8941 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     9001 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     9061 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     9121 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     9181 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     9241 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     9301 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     9361 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     9421 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     9481 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     9541 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     9601 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     9661 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
     9721 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
     9781 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     9841 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     9901 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     9961 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
    10021 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
    10081 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
    10141 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
    10201 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
    10261 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
    10321 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
    10381 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
    10441 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
    10501 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
    10561 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
    10621 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
    10681 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
    10741 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
    10801 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
    10861 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
    10921 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
    10981 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
    11041 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
    11101 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
    11161 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    11221 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
    11281 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
    11341 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
    11401 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
    11461 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
    11521 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
    11581 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
    11641 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
    11701 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
    11761 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
    11821 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
    11881 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
    11941 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
    12001 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
    12061 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    12121 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    12181 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    12241 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
    12301 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    12361 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    12421 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
    12481 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
    12541 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
//
