LOCUS       sequence-325564-        9866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable integration and expression of GCaMP6s in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9866)
  AUTHORS   Chang-Graham AL, Perry JL, Strtak AC, Ramachandran NK, Criglar JM, 
            Philip AA, Patton JT, Estes MK, Hyser JM
  TITLE     Rotavirus Calcium Dysregulation Manifests as Dynamic Calcium 
            Signaling in the Cytoplasm and Endoplasmic Reticulum.
  JOURNAL   Sci Rep. 2019 Jul 25;9(1):10822. doi: 10.1038/s41598-019-46856-8.
  PUBMED    31346185
REFERENCE   2  (bases 1 to 9866)
  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: "Sci Rep."; 
            date: "2019-07-25"; volume: "9(1):10822. doi"; pages: " 
            10.1038/s41598-019-46856-8"
COMMENT     Sequence Label: pLVX-IRES-Hygro_GCaMP6s
FEATURES             Location/Qualifiers
     source          1..9866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2028..2143
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        2201..2504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2659..2679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2705..2729
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2815..2824
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2821..2829,2830..2847,2848..2850,2851..2883,2884..2886,
                     2887..2910,2911..2937,2938..2994,2995..3000,3001..3270,
                     3271..3288,3289..3723,3724..3729,3730..4173)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      2917..2926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3207..3228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3334..3355)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3595..3614)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    join(4190..4764,4765..4767,4768..4776)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4357..4374)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4584..4603
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4802..5821
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             4802..5821
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    5835..6423
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5888..5908)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6306..6317)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6630..7263
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7391..7407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7391..7407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7404..7426)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7415..7431
                     /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(7439..7445,7446..7463,7464..7469))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7484..7505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7622..7639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7793..8381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7873..7892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8552..9343,9344..9412))
                     /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     9175..9194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9413..9517)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9565..9699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9602..9621)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9656..9675
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcct cgaggccacc
     2821 atgggttctc atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg
     2881 ggtcgggatc tgtacgacga tgacgataag gatctcgcca ccatggtcga ctcatcacgt
     2941 cgtaagtgga ataagacagg tcacgcagtc agagctatag gtcggctgag ctcactcgag
     3001 aacgtctata tcaaggccga caagcagaag aacggcatca aggcgaactt ccacatccgc
     3061 cacaacatcg aggacggcgg cgtgcagctc gcctaccact accagcagaa cacccccatc
     3121 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc caaactttcg
     3181 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3241 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat ggtgagcaag
     3301 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3361 ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg ccacctacgg caagctgacc
     3421 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3481 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3541 ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt caaggacgac
     3601 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3661 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3721 aacctgccgg accaactgac tgaagagcag atcgcagaat ttaaagaggc tttctcccta
     3781 tttgacaagg acggggatgg gacaataaca accaaggagc tggggacggt gatgcggtct
     3841 ctggggcaga accccacaga agcagagctg caggacatga tcaatgaagt agatgccgac
     3901 ggtgacggca caatcgactt ccctgagttc ctgacaatga tggcaagaaa aatgaaatac
     3961 agggacacgg aagaagaaat tagagaagcg ttcggtgtgt ttgataagga tggcaatggc
     4021 tacatcagtg cagcagagct tcgccacgtg atgacaaacc ttggagagaa gttaacagat
     4081 gaagaggttg atgaaatgat cagggaagca gacatcgatg gggatggtca ggtaaactac
     4141 gaagagtttg tacaaatgat gacagcgaag tgagcggccg cggatcccgc ccctctccct
     4201 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     4261 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     4321 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     4381 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     4441 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     4501 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     4561 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     4621 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     4681 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     4741 tcctttgaaa aacacgatga taagcttgcc acaacccgta ccaaagatgg atagatccgg
     4801 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     4861 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     4921 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     4981 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     5041 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     5101 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     5161 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     5221 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     5281 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     5341 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     5401 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     5461 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     5521 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     5581 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     5641 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     5701 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     5761 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     5821 gacgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5881 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     5941 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     6001 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     6061 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     6121 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     6181 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     6241 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     6301 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     6361 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     6421 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa
     6481 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     6541 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     6601 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
     6661 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta
     6721 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
     6781 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
     6841 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag
     6901 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
     6961 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
     7021 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta
     7081 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     7141 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     7201 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7261 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     7321 cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc tagagcttgg
     7381 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7441 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     7501 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7561 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7621 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7681 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7741 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7801 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7861 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7921 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7981 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8041 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8101 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8161 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8221 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8281 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8341 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8401 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8461 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8521 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8581 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8641 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8701 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8761 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8821 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8881 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8941 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9001 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9061 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9121 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9181 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9241 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9301 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9361 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9421 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9481 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9541 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa
     9601 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     9661 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     9721 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     9781 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     9841 taaatatgta ctacaaactt agtagt
//
