pHEFI-OptoCARCD19-mScarlet

LOCUS       pHEFI-OptoCARCD19-mScarlet              12575 bp ds-DNA     circular SYN 04-AUG-2021
DEFINITION  Expresses an optically-controllable Chimeric antigen receptor that 
            binds CD19 antigen, fused to mScarlet fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    pHEFI-OptoCARCD19-mScarlet
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12575)
  AUTHORS   Harris MJ, Fuyal M, James JR
  TITLE     Quantifying persistence in the T-cell signaling network using an 
            optically controllable antigen receptor.
  JOURNAL   Mol Syst Biol. 2021 May;17(5):e10091. doi: 10.15252/msb.202010091.
  PUBMED    33988299
REFERENCE   2  (bases 1 to 12575)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..12575
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(25..43)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(26..43)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(333..352)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(566..585)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     685..707
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(745..763)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        831..935
                     /gene="bla"
                     /label=AmpR promoter
     CDS             936..1796
                     /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     complement(1154..1173)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1967..2555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2456..2475
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2709..2726
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2796..2816)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2801..3130
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2981..3116
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3043..3062
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     intron          4264..4329
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4459..4479
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4904..5038
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4941..4960)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4995..5014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5206..5839
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5886..6011
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6508..6741
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6926..6970
                     /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    7255..7372
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7433..8611
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7664..8602
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8559..8579
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      8624..8633
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             8693..8869
                     /codon_start=1
                     /product="""Zdark"" protein, derived from the Z subunit of 
                     protein A, that binds to the dark state of LOV2 and is 
                     released upon irradiation (Wang et al., 2016)"
                     /label=Zdk1
                     /translation="MVDNKFNKEKTRAGAEIHSLPNLNVEQKFAFIVSLFDDPSQSANL
                     LAEAKKLNDAQAPK"
     regulatory      9206..9215
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             9212..9907
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(9345..9363)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9613..9632
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             9929..9985
                     /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"
     misc_feature    11644..12232
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11697..11717)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12115..12126)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12235..12251)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12236..12252)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12342..12575
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 gcatctagaa ttaattccgt gtattctata gtgtcaccta aatcgtatgt gtatgataca
       61 taaggttatg tattaattgt agccgcgttc taacgacaat atgtacaagc ctaattgtgt
      121 agcatctggc ttactgaagc agaccctatc atctctctcg taaactgccg tcagagtcgg
      181 tttggttgga cgaaccttct gagtttctgg taacgccgtc ccgcacccgg aaatggtcag
      241 cgaaccaatc agcagggtca tcgctagcca gatcctctac gccggacgca tcgtggccgg
      301 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
      361 agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg
      421 ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc
      481 ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg
      541 agagcgtcga atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
      601 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
      661 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
      721 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
      781 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
      841 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
      901 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
      961 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     1021 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     1081 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     1141 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     1201 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     1261 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     1321 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     1381 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     1441 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     1501 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1561 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1621 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1681 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1741 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     1801 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     1861 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     1921 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1981 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2041 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2101 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2161 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2221 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2281 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2341 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2401 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2461 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2521 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2581 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2641 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2701 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2761 tccccgcgcg ttggccgatt cattaatgca gctgtggaat gtgtgtcagt tagggtgtgg
     2821 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2881 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2941 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3001 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3061 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3121 cttttgcaaa aagcttggac acaagacagg cttgcgagat atgtttgaga ataccacttt
     3181 atcccgcgtc agggagaggc agtgcgtaaa aagacgcgga ctcatgtgaa atactggttt
     3241 ttagtgcgcc agatctctat aatctcgcgc aacctatttt cccctcgaac actttttaag
     3301 ccgtagataa acaggctggg acacttcaca tgagcgaaaa atacatcgtc acctgggaca
     3361 tgttgcagat ccatgcacgt aaactcgcaa gccgactgat gccttctgaa caatggaaag
     3421 gcattattgc cgtaagccgt ggcggtctgt accgggtgcg ttactggcgc gtgaactggg
     3481 tattcgtcat gtcgataccg tttgtatttc cagctacgat cacgacaacc agcgcgagct
     3541 taaagtgctg aaacgcgcag aaggcgatgg cgaaggcttc atcgttattg atgacctggt
     3601 ggataccggt ggtactgcgg ttgcgattcg tgaaatgtat ccaaaagcgc actttgtcac
     3661 catcttcgca aaaccggctg gtcgtccgct ggttgatgac tatgttgttg atatcccgca
     3721 agatacctgg attgaacagc cgtgggatat gggcgtcgta ttcgtcccgc caatctccgg
     3781 tcgctaatct tttcaacgcc tggcactgcc gggcgttgtt ctttttaact tcaggcgggt
     3841 tacaatagtt tccagtaagt attctggagg ctgcatccat gacacaggca aacctgagcg
     3901 aaaccctgtt caaaccccgc tttaaacatc ctgaaacctc gacgctagtc cgccgcttta
     3961 atcacggcgc acaaccgcct gtgcagtcgg cccttgatgg taaaaccatc cctcactggt
     4021 atcgcatgat taaccgtctg atgtggatct ggcgcggcat tgacccacgc gaaatcctcg
     4081 acgtccaggc acgtattgtg atgagcgatg ccgaacgtac cgacgatgat ttatacgata
     4141 cggtgattgg ctaccgtggc ggcaactgga tttatgagtg ggccccggat ctttgtgaag
     4201 gaaccttact tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct
     4261 aaggtaaata taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt
     4321 gtattttaga ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat
     4381 gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac
     4441 tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct
     4501 tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt
     4561 gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat
     4621 tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt ttttcttact
     4681 ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc
     4741 tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat
     4801 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     4861 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4921 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4981 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
     5041 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac
     5101 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt
     5161 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagttggaa gggctaattc
     5221 actcccaaag aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     5281 ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc tttggatggt
     5341 gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa ggagagaaca
     5401 ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtgttag
     5461 agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg catccggagt
     5521 acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg gactttccag
     5581 ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc atataagcag
     5641 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5701 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5761 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5821 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     5881 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     5941 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     6001 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     6061 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     6121 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     6181 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     6241 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     6301 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccggccgct
     6361 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     6421 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     6481 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     6541 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     6601 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     6661 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     6721 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     6781 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     6841 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     6901 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     6961 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     7021 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     7081 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     7141 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     7201 tcgattagtg aacggatctc gacggtatcg ccaaatggca gtattcatcc acaattttaa
     7261 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     7321 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
     7381 ttacagggac agcagagatc cagtttggat cgataagctt gatatcgaat tcggctccgg
     7441 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     7501 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     7561 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     7621 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
     7681 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
     7741 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     7801 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
     7861 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     7921 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     7981 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     8041 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     8101 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     8161 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     8221 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     8281 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     8341 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     8401 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     8461 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     8521 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     8581 aagttttttt cttccatttc aggtgtcgtg aggaattgcg atcgccacca tgggctgtgg
     8641 ctgcagctca cacccggaag atgacggcgg atctggaggc tccggcggat ctatggtgga
     8701 taacaaattc aataaagaaa agacacgtgc cggtgcggaa atccattctc tgccaaacct
     8761 gaatgttgag cagaagtttg ccttcatcgt gagcctgttt gatgatccat ctcagagcgc
     8821 caatctgctg gccgaagcca aaaaactgaa cgatgcccag gccccaaaaa ctagcgcaga
     8881 cgcccccgcg taccagcagg gccagaacca gctctataac gagctcaatc taggacgaag
     8941 agaggagtac gatgttttgg acaagagacg tggccgggac cctgagatgg ggggaaagcc
     9001 gcagagaagg aagaaccctc aggaaggcct gtacaatgaa ctgcagaaag ataagatggc
     9061 ggaggcctac agtgagattg ggatgaaagg cgagcgccgg aggggcaagg ggcacgatgg
     9121 cctttaccag ggtctcagta cagccaccaa ggacacctac gacgcccttc acatgcaggc
     9181 cctgccccct cgggatccac cggtcgccac catggtgagc aagggcgagg cagtgatcaa
     9241 ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg agttcgagat
     9301 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     9361 caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca tgtacggctc
     9421 cagggccttc accaagcacc ccgccgacat ccccgactac tataagcagt ccttccccga
     9481 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca
     9541 ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg gcaccaactt
     9601 ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg
     9661 gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc gcctgaagga
     9721 cggaggccgc tacctggcgg acttcaagac cacctacaag gccaagaagc ccgtgcagat
     9781 gcccggcgcc tacaacgtcg accgcaagtt ggacatcacc tcccacaacg aggactacac
     9841 cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca tggacgagct
     9901 gtacaagacc agttaccagg gccctggcgc cacgaacttc tctctgttaa agcaagcagg
     9961 agacgtggaa gaaaaccccg gtcccacgcg tatggcctta ccagtgaccg ccttgctcct
    10021 gccgctggcc ttgctgctcc acgccgccag gccggacatc cagatgacac agactacatc
    10081 ctccctgtct gcctctctgg gagacagagt caccatcagt tgcagggcaa gtcaggacat
    10141 tagtaaatat ttaaattggt atcagcagaa accagatgga actgttaaac tcctgatcta
    10201 ccatacatca agattacact caggagtccc atcaaggttc agtggcagtg ggtctggaac
    10261 agattattct ctcaccatta gcaacctgga gcaagaagat attgccactt acttttgcca
    10321 acagggtaat acgcttccgt acacgttcgg aggggggacc aagctggaga tcacaggtgg
    10381 cggtggctcg ggcggtggtg ggtcgggtgg cggcggatct gaggtgaaac tgcaggagtc
    10441 aggacctggc ctggtggcgc cctcacagag cctgtccgtc acatgcactg tctcaggggt
    10501 ctcattaccc gactatggtg taagctggat tcgccagcct ccacgaaagg gtctggagtg
    10561 gctgggagta atatggggta gtgaaaccac atactataat tcagctctca aatccagact
    10621 gaccatcatc aaggacaact ccaagagcca agttttctta aaaatgaaca gtctgcaaac
    10681 tgatgacaca gccatttact actgtgccaa acattattac tacggtggta gctatgctat
    10741 ggactactgg ggccaaggaa cctcagtcac cgtctcctca accacgacgc cagcgccgcg
    10801 accaccaaca ccggcgccca ccatcgcgtc gcagcccctg tccctgcgcc cagaggcgtg
    10861 ccggccagcg gcggggggcg cagtgcacac gagggggctg gacttcgcct gtgatatcta
    10921 catctgggcg cccttggccg ggacttgtgg ggtccttctc ctgtcactgg ttatcaccct
    10981 ttactgcaaa cggggcagaa agaaactcct gtatatattc aaacaaccat ttatgagacc
    11041 agtacaaact actcaagagg aagatggctg tagctgccga tttccagaag aagaagaagg
    11101 aggatgtgaa ctgagagtga agttcagcag gactagtggc ggaagcggaa gtttggctac
    11161 tacacttgaa cgtattgaga agaactttct cattactgac ccaagattgc cagataatcc
    11221 cattatattc gcgtccgata gtttcttgca gttgacagaa tatagccgtg aagaaatttt
    11281 gggaagaaac tgcaggtttc tacaaggtcc tgaaactgat cgcgcgacag tgagaaaaat
    11341 tagagatgcc atagataacc aaacagaggt cactgttcag ctgattaatt atacaaagag
    11401 tggtaaaaag ttctggaacc tctttcactt gcagcctatg cgagatcaga agggagatgt
    11461 ccagtacttt attggggttc agttggatgg aactgagcat gtccgagatg ctgccgagag
    11521 agagggagtc atgctgatta agaaaactgc agaaaatatt gatgaggcgg caaaagaact
    11581 ttgagcggcc gcgactctag agtcgacctg caggcatgca agcttgatat caagcttatc
    11641 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
    11701 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
    11761 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
    11821 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
    11881 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
    11941 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
    12001 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
    12061 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
    12121 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
    12181 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
    12241 cgtcgacctc gagggaatta attcgagctc ggtaccttta agaccaatga cttacaaggc
    12301 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
    12361 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
    12421 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
    12481 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
    12541 cagacccttt tagtcagtgt ggaaaatctc tagca
//