LOCUS       sequence-176273-       13990 bp    DNA     circular SYN 15-APR-2025
DEFINITION  MESA protease chain with V1-MESA ectodomain, 45 extracellular 
            linkers, a flag tag, and Tev protease, MS2-P65-HSF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13990)
  AUTHORS   Schwarz KA, Daringer NM, Dolberg TB, Leonard JN
  TITLE     Rewiring human cellular input-output using modular extracellular 
            sensors.
  JOURNAL   Nat Chem Biol. 2016 Dec 12. doi: 10.1038/nchembio.2253.
  PUBMED    27941759
REFERENCE   2  (bases 1 to 13990)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem 
            Biol. 2016 Dec 12. doi: 10.1038/nchembio.2253."
COMMENT     Sequence Label: V1-MESA-45F-Tev-MS2-p65-HSF1
FEATURES             Location/Qualifiers
     source          1..13990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(110..127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(281..869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(361..380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1040..1831,1832..1900))
                     /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     1663..1682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1901..2005)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2053..2187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             2355..2988
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3035..3160
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3657..3890
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4075..4119
                     /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    4417..4534
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(4599..4973)
                     /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"
     regulatory      4970..4979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4983..5001)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4993..5040)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     enhancer        5145..5448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5449..5652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5603..5623
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5649..5673
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6723..7427
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIV
                     GIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEE
                     PFQPVKEATQLMN"
     primer_bind     7706..7726
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7765..7774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7774..8160
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             8215..8235
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8251..8793
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             8818..9189
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             9205..9258
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9262..10281
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     misc_feature    10321..10909
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10374..10394)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10792..10803)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10912..10928)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10913..10929)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11114..11347
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11448..11465)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11454..11678
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11724..12152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11811..11830)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12021..12042
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12161..12181)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12166..12495
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12346..12481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12408..12427
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             12544..13569
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             12544..13569
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    13699..13820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13869..13885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13869..13885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13882..13904)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13893..13909
                     /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(13917..13923,13924..13941,13942..13947))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13962..13983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
       61 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
      121 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      181 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      241 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      301 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      361 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      421 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      481 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      541 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      601 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      661 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
      721 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      781 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      841 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      901 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      961 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1021 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1081 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1141 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1201 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1261 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1321 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1381 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1441 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1501 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1561 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1621 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1681 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1741 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1801 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1861 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1921 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1981 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     2041 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2101 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2161 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     2221 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     2281 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     2341 acaaacttag tagttggaag ggctaattca ctcccaaaga agacaagata tccttgatct
     2401 gtggatctac cacacacaag gctacttccc tgattagcag aactacacac cagggccagg
     2461 ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa
     2521 ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg
     2581 gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca
     2641 tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta
     2701 caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg
     2761 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
     2821 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2881 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2941 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     3001 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
     3061 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
     3121 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
     3181 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
     3241 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
     3301 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
     3361 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
     3421 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
     3481 gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg
     3541 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     3601 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     3661 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     3721 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     3781 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     3841 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     3901 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     3961 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     4021 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     4081 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     4141 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     4201 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     4261 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     4321 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
     4381 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     4441 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     4501 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     4561 tccagtttgg ttagtaccgg gcccgctcta gtccggaatc agtcctgctc ctcggccacg
     4621 aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg
     4681 ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac
     4741 cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc
     4801 accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg
     4861 aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct
     4921 ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtggcc
     4981 ctcctatagt gagtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac
     5041 acgtgctgca ggtccgaggt tctagacgta ttaccgccat gcattagtta ttaatagtaa
     5101 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     5161 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     5221 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     5281 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     5341 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     5401 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     5461 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     5521 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     5581 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     5641 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     5701 gacctccata gaagacaccg actctactag aggatctgcc accagctagc gccatcatgg
     5761 cttggaccag cttaattctt agcttgcttg ccctgtgcag cggtgcatca tctgacatac
     5821 agatgactca gtcacctagc agtctgtctg cttcagtcgg tgatagggta actatcacct
     5881 gtcgggcatc tcaggatgtg tccacggcag ttgcctggta ccagcagaag cccggcaagg
     5941 cgcctaagtt gcttatctat tcagcttcat ttttgtatag tggggtgcct agccggttta
     6001 gcggaagtgg ttctggcaca gacttcactt tgacaatcag ttccctccaa ccggaggatt
     6061 tcgctaccta ctactgtcag caggggtacg gaaacccctt tacgtttgga cagggcacta
     6121 aagtggagat taagagggga ggcggaggct ccggtggggg aggctctggt ggtggaggta
     6181 gcgaggttca gctggtggaa agtggtggcg gactagtaca gccaggcggc tccctgcggc
     6241 tgagttgcgc cgcgagtggc tttactatat ctgactattg gatccattgg gtcaggcaag
     6301 cccctgggaa aggattggag tgggttgctg gtattacccc cgcaggtggg tatacttatt
     6361 atgcagatag cgtgaaaggg aggttcacca tctctgcaga cacaagcaag aacacggcct
     6421 acctacagat gaactctctg agggcagagg acactgccgt ctactattgc gcccgatttg
     6481 tgttttttct accctacgcc atggactatt ggggacaggg gacattggta ggcgccggga
     6541 caggagggtc tgggggtggc actggcggat ctggaggaga ctacaaggac gatgacgaca
     6601 acggcggatc gggggggtct ggtggtagtg gcggttcggg gggaggtacc ggtctggtcg
     6661 tggttgctgg agtcctgttt tgttatggct tgctagtgac agtggctctt tgtgttgaaa
     6721 acgagagctt gtttaagggg ccgcgtgatt acaacccgat atcgagcacc atttgtcatt
     6781 tgacgaatga atctgatggg cacacaacat cgttgtatgg tattggattt ggtcccttca
     6841 tcattacaaa caagcacttg tttagaagaa ataatggaac actgttggtc caatcactac
     6901 atggtgtatt caaggtcaag aacaccacga ctttgcaaca acacctcatt gatgggaggg
     6961 acatgataat tattcgcatg cctaaggatt tcccaccatt tcctcaaaag ctgaaattta
     7021 gagagccaca aagggaagag cgcatatgtc ttgtgacaac caacttccaa actaagagca
     7081 tgtctagcat ggtgtcagac actagttgca cattcccttc atctgatggc atattctgga
     7141 agcattggat tcaaaccaag gatgggcagt gtggcagtcc attagtatca actagagatg
     7201 ggttcattgt tggtatacac tcagcatcga atttcaccaa cacaaacaat tatttcacaa
     7261 gcgtgccgaa aaacttcatg gaattgttga caaatcagga ggcgcagcag tgggttagtg
     7321 gttggcgatt aaatgctgac tcagtattgt gggggggcca taaagttttc atggtgaaac
     7381 ctgaagagcc ttttcagcca gttaaggaag cgactcaact catgaattag ggcgcgccgc
     7441 tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg
     7501 tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
     7561 cagtccccga gaagttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     7621 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     7681 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     7741 ccatttcagg tgtcgtgacg tacggccacc atggcttcaa actttactca gttcgtgctc
     7801 gtggacaatg gtgggacagg ggatgtgaca gtggctcctt ctaatttcgc taatggggtg
     7861 gcagagtgga tcagctccaa ctcacggagc caggcctaca aggtgacatg cagcgtcagg
     7921 cagtctagtg cccagaagag aaagtatacc atcaaggtgg aggtccccaa agtggctacc
     7981 cagacagtgg gcggagtcga actgcctgtc gccgcttgga ggtcctacct gaacatggag
     8041 ctcactatcc caattttcgc taccaattct gactgtgaac tcatcgtgaa ggcaatgcag
     8101 gggctcctca aagacggtaa tcctatccct tccgccatcg ccgctaactc aggtatctac
     8161 agcgctggag gaggtggaag cggaggagga ggaagcggag gaggaggtag cggacctaag
     8221 aaaaagagga aggtggcggc cgctggatcc ccttcagggc agatcagcaa ccaggccctg
     8281 gctctggccc ctagctccgc tccagtgctg gcccagacta tggtgccctc tagtgctatg
     8341 gtgcctctgg cccagccacc tgctccagcc cctgtgctga ccccaggacc accccagtca
     8401 ctgagcgctc cagtgcccaa gtctacacag gccggcgagg ggactctgag tgaagctctg
     8461 ctgcacctgc agttcgacgc tgatgaggac ctgggagctc tgctggggaa cagcaccgat
     8521 cccggagtgt tcacagatct ggcctccgtg gacaactctg agtttcagca gctgctgaat
     8581 cagggcgtgt ccatgtctca tagtacagcc gaaccaatgc tgatggagta ccccgaagcc
     8641 attacccggc tggtgaccgg cagccagcgg ccccccgacc ccgctccaac tcccctggga
     8701 accagcggcc tgcctaatgg gctgtccgga gatgaagact tctcaagcat cgctgatatg
     8761 gactttagtg ccctgctgtc acagatttcc tctagtgggc agggaggagg tggaagcggc
     8821 ttcagcgtgg acaccagtgc cctgctggac ctgttcagcc cctcggtgac cgtgcccgac
     8881 atgagcctgc ctgaccttga cagcagcctg gccagtatcc aagagctcct gtctccccag
     8941 gagcccccca ggcctcccga ggcagagaac agcagcccgg attcagggaa gcagctggtg
     9001 cactacacag cgcagccgct gttcctgctg gaccccggct ccgtggacac cgggagcaac
     9061 gacctgccgg tgctgtttga gctgggagag ggctcctact tctccgaagg ggacggcttc
     9121 gccgaggacc ccaccatctc cctgctgaca ggctcggagc ctcccaaagc caaggacccc
     9181 actgtctcct gtacaggcag tggagagggc agaggaagtc tgctaacatg cggtgacgtc
     9241 gaggagaatc ctggcccaac catgaaaaag cctgaactca ccgctacctc tgtcgagaag
     9301 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctccga gggcgaagaa
     9361 tctcgggctt tcagcttcga tgtgggaggg cgtggatatg tcctgcgggt gaatagctgc
     9421 gccgatggtt tctacaaaga tcgctatgtt tatcggcact ttgcatccgc cgctctccct
     9481 attcccgaag tgcttgacat tggggagttc agcgagagcc tgacctattg catctcccgc
     9541 cgtgcacagg gtgtcacctt gcaagacctg cctgaaaccg aactgcccgc tgttctccag
     9601 cccgtcgccg aggccatgga tgccatcgct gccgccgatc ttagccagac cagcgggttc
     9661 ggcccattcg gacctcaagg aatcggtcaa tacactacat ggcgcgattt catctgcgct
     9721 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcctcc
     9781 gtcgcccagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     9841 ctcgtgcacg ccgatttcgg ctccaacaat gtcctgaccg acaatggccg cataacagcc
     9901 gtcattgact ggagcgaggc catgttcggg gattcccaat acgaggtcgc caacatcttc
     9961 ttctggaggc cctggttggc ttgtatggag cagcagaccc gctacttcga gcggaggcat
    10021 cccgagcttg caggatctcc tcggctccgg gcttatatgc tccgcattgg tcttgaccaa
    10081 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggctca gggtcgctgc
    10141 gacgcaatcg tccggtccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
    10201 gctgccgtct ggaccgatgg ctgtgtggaa gtgctcgccg atagtggaaa cagacgcccc
    10261 agcactcgtc ctagggcaaa ggatctgcag taatgagaat tcgatatcaa gcttatcggt
    10321 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
    10381 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
    10441 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
    10501 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
    10561 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
    10621 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
    10681 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
    10741 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
    10801 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
    10861 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
    10921 cgacctcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
    10981 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
    11041 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
    11101 gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
    11161 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
    11221 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
    11281 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
    11341 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
    11401 atatcagaga gtgagaggcc ttgacattgt ttaaacccgc tgatcagcct cgactgtgcc
    11461 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
    11521 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
    11581 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
    11641 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
    11701 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
    11761 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
    11821 tttcttccct tcctttctcg tcacgttcgc cggctttccc cgtcaagctc taaatcgggg
    11881 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    11941 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    12001 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
    12061 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    12121 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
    12181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    12241 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    12301 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    12361 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    12421 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
    12481 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
    12541 gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
    12601 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
    12661 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
    12721 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
    12781 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
    12841 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
    12901 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
    12961 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
    13021 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
    13081 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
    13141 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
    13201 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
    13261 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
    13321 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
    13381 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
    13441 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
    13501 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
    13561 aaggaatagc acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
    13621 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
    13681 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    13741 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    13801 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
    13861 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    13921 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    13981 ttaattgcgt
//
