LOCUS       sequence-99988-e       11409 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the MS2-P65-HSF1 activator helper complex with a 2A GFP. 
            3rd generation lentiviral vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11409)
  AUTHORS   Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena 
            C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F
  TITLE     Genome-scale transcriptional activation by an engineered CRISPR-Cas9
            complex.
  JOURNAL   Nature. 2014 Dec 10. doi: 10.1038/nature14136.
  PUBMED    25494202
REFERENCE   2  (bases 1 to 11409)
  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: "Nature. 
            2014 Dec 10. doi: 10.1038/nature14136."
COMMENT     Sequence Label: MS2-P65-HSF1_GFP
FEATURES             Location/Qualifiers
     source          1..11409
                     /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        2682..3859
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2913..3850
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3807..3827
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3866..3875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3875..4261
                     /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             4316..4336
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4352..4894
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             4919..5290
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             5306..5359
                     /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             join(5360..5362,5363..6076)
                     /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(5360..5362,5363..6073)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5402..5423)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5663..5682)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6010..6031
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6101..6689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6154..6174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6572..6583)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6692..6708)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6693..6709)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7214..7394
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7420..7437)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7426..7650
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7696..8124
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7783..7802)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7993..8014
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8133..8153)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8138..8467
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8318..8453
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8380..8399
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8515..8562
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8581..8955
                     /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    9085..9206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9122..9141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9176..9195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9255..9271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9255..9271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9268..9290)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9279..9295
                     /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(9303..9309,9310..9327,9328..9333))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9348..9369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9486..9503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9657..10245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9737..9756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10416..11207,11208..11276))
                     /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     11039..11058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11277..11381)
                     /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 atgcaaagat ggataaagtt ttaaacagag aggaatcttt
     2641 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt
     2701 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2761 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2821 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2881 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2941 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccact ggctgcagta
     3001 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     3061 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     3121 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     3181 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     3241 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     3301 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     3361 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     3421 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     3481 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     3541 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     3601 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     3661 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     3721 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     3781 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     3841 tccatttcag gtgtcgtgac gtacggccac catggcttca aactttactc agttcgtgct
     3901 cgtggacaat ggtgggacag gggatgtgac agtggctcct tctaatttcg ctaatggggt
     3961 ggcagagtgg atcagctcca actcacggag ccaggcctac aaggtgacat gcagcgtcag
     4021 gcagtctagt gcccagaaga gaaagtatac catcaaggtg gaggtcccca aagtggctac
     4081 ccagacagtg ggcggagtcg aactgcctgt cgccgcttgg aggtcctacc tgaacatgga
     4141 gctcactatc ccaattttcg ctaccaattc tgactgtgaa ctcatcgtga aggcaatgca
     4201 ggggctcctc aaagacggta atcctatccc ttccgccatc gccgctaact caggtatcta
     4261 cagcgctgga ggaggtggaa gcggaggagg aggaagcgga ggaggaggta gcggacctaa
     4321 gaaaaagagg aaggtggcgg ccgctggatc cccttcaggg cagatcagca accaggccct
     4381 ggctctggcc cctagctccg ctccagtgct ggcccagact atggtgccct ctagtgctat
     4441 ggtgcctctg gcccagccac ctgctccagc ccctgtgctg accccaggac caccccagtc
     4501 actgagcgct ccagtgccca agtctacaca ggccggcgag gggactctga gtgaagctct
     4561 gctgcacctg cagttcgacg ctgatgagga cctgggagct ctgctgggga acagcaccga
     4621 tcccggagtg ttcacagatc tggcctccgt ggacaactct gagtttcagc agctgctgaa
     4681 tcagggcgtg tccatgtctc atagtacagc cgaaccaatg ctgatggagt accccgaagc
     4741 cattacccgg ctggtgaccg gcagccagcg gccccccgac cccgctccaa ctcccctggg
     4801 aaccagcggc ctgcctaatg ggctgtccgg agatgaagac ttctcaagca tcgctgatat
     4861 ggactttagt gccctgctgt cacagatttc ctctagtggg cagggaggag gtggaagcgg
     4921 cttcagcgtg gacaccagtg ccctgctgga cctgttcagc ccctcggtga ccgtgcccga
     4981 catgagcctg cctgaccttg acagcagcct ggccagtatc caagagctcc tgtctcccca
     5041 ggagcccccc aggcctcccg aggcagagaa cagcagcccg gattcaggga agcagctggt
     5101 gcactacaca gcgcagccgc tgttcctgct ggaccccggc tccgtggaca ccgggagcaa
     5161 cgacctgccg gtgctgtttg agctgggaga gggctcctac ttctccgaag gggacggctt
     5221 cgccgaggac cccaccatct ccctgctgac aggctcggag cctcccaaag ccaaggaccc
     5281 cactgtctcc gctagcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt
     5341 cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     5401 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     5461 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     5521 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     5581 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     5641 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     5701 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     5761 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     5821 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     5881 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     5941 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     6001 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     6061 cgagctgtac aagtaagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     6121 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     6181 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     6241 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     6301 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     6361 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6421 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6481 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     6541 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6601 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6661 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa
     6721 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     6781 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     6841 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     6901 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     6961 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     7021 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     7081 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     7141 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     7201 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     7261 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     7321 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     7381 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     7441 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     7501 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     7561 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     7621 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     7681 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     7741 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     7801 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     7861 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7921 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7981 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     8041 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     8101 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     8161 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     8221 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     8281 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     8341 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     8401 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     8461 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     8521 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     8581 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     8641 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     8701 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     8761 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     8821 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     8881 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     8941 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     9001 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     9061 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     9121 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     9181 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     9241 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     9301 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     9361 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     9421 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     9481 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     9541 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     9601 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     9661 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     9721 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     9781 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     9841 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     9901 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     9961 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
    10021 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
    10081 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
    10141 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
    10201 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    10261 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    10321 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    10381 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    10441 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    10501 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    10561 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
    10621 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    10681 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    10741 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    10801 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    10861 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    10921 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    10981 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    11041 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    11101 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    11161 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    11221 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    11281 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    11341 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    11401 ccacctgac
//
