LOCUS       Lenti_MS2_P65_VP       11316 bp ds-DNA     circular     02-OCT-2018
DEFINITION  lenti vector encoding the MS2-P65-HSF1 activator helper complex
            with a 2A Hygro resistance marker NOTE: A version of this plasmid
            with  improved titer is available: Addgene plasmid #89308 lentiMPH
            v2.
ACCESSION   .
VERSION     .
KEYWORDS    lenti MS2-P65-HSF1_Hygro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11736)
  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 11736)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     source          1..5368
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(47..66)
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
                     /label=pRS-marker
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     enhancer        238..617
                     /note="human cytomegalovirus immediate early enhancer"
                     /label=CMV enhancer
                     /ApEinfo_fwdcolor="#5ac3fa"
                     /ApEinfo_revcolor="#5ac3fa"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        619..817
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label=CMV promoter
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     772..792
                     /note="Human CMV immediate early promoter, forward primer"
                     /label=CMV-F
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     LTR             835..1015
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label=5' LTR (truncated)
                     /ApEinfo_fwdcolor="#86f71d"
                     /ApEinfo_revcolor="#d7336f"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1062..1187
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label=HIV-1 Psi
                     /ApEinfo_fwdcolor="#fff0e8"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    1680..1913
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label=RRE
                     /ApEinfo_fwdcolor="#fff0e8"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2440..2557
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label=cPPT/CTS
                     /ApEinfo_fwdcolor="#fff0e8"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        2687..3865
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
                     /label=EF-1-alpha promoter
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     intron          2918..3856
                     /note="intron upstream of the start codon of human
                     EF-1-alpha"
                     /label=EF-1-alpha intron A
                     /ApEinfo_fwdcolor="#eb606c"
                     /ApEinfo_revcolor="#eb606c"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     3813..3833
                     /note="Human elongation factor-1a promoter, forward
                     primer"
                     /label=EF1a-F
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    5369..5968
                     /vntifkey="21"
                     /label=puro
                     /ApEinfo_fwdcolor="#78a79f"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     regulatory      3872..3881
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /label=vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             3881..4267
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /note="binds with high affinity to a specific stem-loop
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKV
                     T
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCEL
                     I VKAMQGLLKDGNPIPSAIAANSGIY"
                     /label=MS2-N55K
                     /ApEinfo_fwdcolor="#2413ff"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             4322..4342
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="PKKKRKV"
                     /label=SV40 NLS
                     /ApEinfo_fwdcolor="#2413ff"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             4358..4900
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse
                     NF-kappa-B"
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLT
                     P
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNS
                     E
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDE
                     D FSSIADMDFSALLSQISS"
                     /label=p65
                     /ApEinfo_fwdcolor="#2413ff"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             4925..5296
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat
                     shock transcription factor HSF1"
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPP
                     R
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAE
                     D PTISLLTGSEPPKAKDPTVS"
                     /label=HSF1
                     /ApEinfo_fwdcolor="#2413ff"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             5312..5365
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label=T2A
                     /ApEinfo_fwdcolor="#2413ff"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    6008..6596
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
                     /label=WPRE
                     /ApEinfo_fwdcolor="#fff0e8"
                     /ApEinfo_revcolor="#7eff74"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(6061..6081)
                     /note="WPRE, reverse primer"
                     /label=WPRE-R
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(6479..6490)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
                     /label=Factor Xa site
                     /ApEinfo_fwdcolor="#2413ff"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(6599..6615)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label=KS primer
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(6600..6616)
                     /note="For pBluescript vector"
                     /label=pBluescriptKS
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     LTR             7121..7301
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label=5' LTR (truncated)(1)
                     /ApEinfo_label="5' LTR (truncated)"
                     /ApEinfo_fwdcolor="#86f71d"
                     /ApEinfo_revcolor="#d7336f"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(7327..7344)
                     /note="Bovine growth hormone terminator, reverse primer.
                     Also called BGH reverse"
                     /label=BGH-rev
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     polyA_signal    7333..7557
                     /note="bovine growth hormone polyadenylation signal"
                     /label=bGH poly(A) signal
                     /ApEinfo_fwdcolor="#ff3eee"
                     /ApEinfo_revcolor="#ff3eee"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      7603..8031
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label=f1 ori
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(7690..7709)
                     /note="F1 origin, reverse primer"
                     /label=F1ori-R
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     7900..7921
                     /note="F1 origin, forward primer"
                     /label=F1ori-F
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(8040..8060)
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
                     /label=pBABE 3'
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        8045..8374
                     /note="SV40 enhancer and early promoter"
                     /label=SV40 promoter
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      8225..8360
                     /note="SV40 origin of replication"
                     /label=SV40 ori
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     8287..8306
                     /note="SV40 promoter/origin, forward primer"
                     /label=SV40pro-F
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        8422..8469
                     /note="synthetic bacterial promoter "
                     /label=EM7 promoter
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             8488..8862
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD
                     D
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWG
                     R EFALRDPAGNCVHFVAEEQD"
                     /label=BleoR
                     /ApEinfo_fwdcolor="#2413ff"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     polyA_signal    8992..9113
                     /note="SV40 polyadenylation signal"
                     /label=SV40 poly(A) signal
                     /ApEinfo_fwdcolor="#ff3eee"
                     /ApEinfo_revcolor="#ff3eee"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(9029..9048)
                     /note="SV40 polyA, reverse primer"
                     /label=SV40pA-R
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     9083..9102
                     /note="SV40 polyA terminator, reverse primer"
                     /label=EBV-rev
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(9162..9178)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label=M13 rev
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(9162..9178)
                     /note="In lacZ gene. Also called M13-rev"
                     /label=M13 Reverse
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(9175..9197)
                     /note="In lacZ gene"
                     /label=M13/pUC Reverse
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    9186..9202
                     /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)."
                     /label=lac operator
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        complement(9210..9240)
                     /note="promoter for the E. coli lac operon"
                     /label=lac promoter
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     protein_bind    9255..9276
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label=CAP binding site
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(9393..9410)
                     /note="L4440 vector, forward primer"
                     /label=L4440
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     rep_origin      complement(9564..10152)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label=ori
                     /ApEinfo_fwdcolor="#999999"
                     /ApEinfo_revcolor="#999999"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     complement(9644..9663)
                     /note="pBR322 origin, forward primer"
                     /label=pBR322ori-F
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             complement(10323..11183)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     I
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     S
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     W
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     A
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     S LIKHW"
                     /label=AmpR
                     /ApEinfo_fwdcolor="#2413ff"
                     /ApEinfo_revcolor="#e9d024"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     primer_bind     10946..10965
                     /note="Ampicillin resistance gene, reverse primer"
                     /label=Amp-R
                     /ApEinfo_fwdcolor="#14c0bd"
                     /ApEinfo_revcolor="#4ec02b"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     promoter        complement(11184..11288)
                     /gene="bla"
                     /label=AmpR promoter
                     /ApEinfo_fwdcolor="#346ee0"
                     /ApEinfo_revcolor="#346ee0"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     source          5969..11316
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct(1)
                     /ApEinfo_label="source:synthetic DNA construct"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
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 gctagctgca aagatggata aagttttaaa cagagaggaa
     2641 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     2701 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     2761 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2821 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     2881 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
     2941 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
     3001 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
     3061 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
     3121 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
     3181 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
     3241 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
     3301 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
     3361 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
     3421 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     3481 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tgaaggacgc ggcgctcggg
     3541 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     3601 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     3661 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     3721 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     3781 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     3841 ttttcttcca tttcaggtgt cgtgacgtac ggccaccatg gcttcaaact ttactcagtt
     3901 cgtgctcgtg gacaatggtg ggacagggga tgtgacagtg gctccttcta atttcgctaa
     3961 tggggtggca gagtggatca gctccaactc acggagccag gcctacaagg tgacatgcag
     4021 cgtcaggcag tctagtgccc agaagagaaa gtataccatc aaggtggagg tccccaaagt
     4081 ggctacccag acagtgggcg gagtcgaact gcctgtcgcc gcttggaggt cctacctgaa
     4141 catggagctc actatcccaa ttttcgctac caattctgac tgtgaactca tcgtgaaggc
     4201 aatgcagggg ctcctcaaag acggtaatcc tatcccttcc gccatcgccg ctaactcagg
     4261 tatctacagc gctggaggag gtggaagcgg aggaggagga agcggaggag gaggtagcgg
     4321 acctaagaaa aagaggaagg tggcggccgc tggatcccct tcagggcaga tcagcaacca
     4381 ggccctggct ctggccccta gctccgctcc agtgctggcc cagactatgg tgccctctag
     4441 tgctatggtg cctctggccc agccacctgc tccagcccct gtgctgaccc caggaccacc
     4501 ccagtcactg agcgctccag tgcccaagtc tacacaggcc ggcgagggga ctctgagtga
     4561 agctctgctg cacctgcagt tcgacgctga tgaggacctg ggagctctgc tggggaacag
     4621 caccgatccc ggagtgttca cagatctggc ctccgtggac aactctgagt ttcagcagct
     4681 gctgaatcag ggcgtgtcca tgtctcatag tacagccgaa ccaatgctga tggagtaccc
     4741 cgaagccatt acccggctgg tgaccggcag ccagcggccc cccgaccccg ctccaactcc
     4801 cctgggaacc agcggcctgc ctaatgggct gtccggagat gaagacttct caagcatcgc
     4861 tgatatggac tttagtgccc tgctgtcaca gatttcctct agtgggcagg gaggaggtgg
     4921 aagcggcttc agcgtggaca ccagtgccct gctggacctg ttcagcccct cggtgaccgt
     4981 gcccgacatg agcctgcctg accttgacag cagcctggcc agtatccaag agctcctgtc
     5041 tccccaggag ccccccaggc ctcccgaggc agagaacagc agcccggatt cagggaagca
     5101 gctggtgcac tacacagcgc agccgctgtt cctgctggac cccggctccg tggacaccgg
     5161 gagcaacgac ctgccggtgc tgtttgagct gggagagggc tcctacttct ccgaagggga
     5221 cggcttcgcc gaggacccca ccatctccct gctgacaggc tcggagcctc ccaaagccaa
     5281 ggaccccact gtctcctgta caggcagtgg agagggcaga ggaagtctgc taacatgcgg
     5341 tgacgtcgag gagaatcctg gcccaaccat gaccgagtac aagcccacgg tgcgcctcgc
     5401 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     5461 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     5521 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     5581 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     5641 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     5701 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     5761 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5821 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5881 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5941 gtgcatgacc cgcaagcccg gtgcctgaga tctgcagtaa tgagaattcg atatcaagct
     6001 tatcggtaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     6061 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     6121 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     6181 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     6241 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     6301 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     6361 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     6421 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     6481 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6541 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     6601 ataccgtcga cctcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
     6661 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
     6721 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6781 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct
     6841 tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg
     6901 gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca
     6961 agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct
     7021 gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc
     7081 atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca
     7141 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     7201 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     7261 atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct
     7321 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     7381 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     7441 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     7501 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     7561 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     7621 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     7681 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7741 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7801 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7861 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7921 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     7981 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     8041 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     8101 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     8161 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     8221 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     8281 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     8341 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     8401 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
     8461 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     8521 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     8581 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     8641 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     8701 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     8761 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     8821 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     8881 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     8941 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     9001 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     9061 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     9121 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     9181 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     9241 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     9301 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     9361 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     9421 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     9481 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     9541 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     9601 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     9661 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     9721 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     9781 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     9841 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     9901 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     9961 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
    10021 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    10081 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    10141 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    10201 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
    10261 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
    10321 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
    10381 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
    10441 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
    10501 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    10561 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    10621 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    10681 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    10741 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    10801 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    10861 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    10921 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    10981 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    11041 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    11101 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    11161 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    11221 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    11281 gttccgcgca catttccccg aaaagtgcca cctgac  
//
