LOCUS       sequence-265944-       10963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For simple determination of the multiplicity of infection (MOI) of a
            lentiviral CRISPR library by checking EGFP expression (still 
            allowing for puro selection)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10963)
  AUTHORS   Panda SK, Wigerblad G, Jiang L, Jimenez-Andrade Y, Iyer VS, Shen Y, 
            Boddul SV, Guerreiro-Cacais AO, Raposo B, Kasza Z, Wermeling F
  TITLE     IL-4 controls activated neutrophil FcgammaR2b expression and 
            migration into inflamed joints.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Jan 24. pii: 1914186117. doi: 
            10.1073/pnas.1914186117.
  PUBMED    31980518
REFERENCE   2  (bases 1 to 10963)
  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: "Proc Natl 
            Acad Sci U S A. 2020 Jan 24. pii: 1914186117. doi: 
            10.1073/pnas.1914186117."
COMMENT     Sequence Label: LentiGuide Puro-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..10963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..1265
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1256
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1213..1233
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1278..1874
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1278..1297)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1774..1794
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1884..1940
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(1941..1943,1944..2657)
                     /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(1941..1943,1944..2654)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1983..2004)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2244..2263)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2591..2612
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2673..3261
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2726..2746)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3144..3155)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3333..3566
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3644..3765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3681..3700)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3735..3754
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3805..3940
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3960..3979)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3961..3979)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3989..4006)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3989..4005)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3998..4020)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4147..4602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4234..4253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4444..4465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4628..4732
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4733..4801,4802..5593)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4951..4970)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5764..6352
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6253..6272
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6506..6523
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6640..6661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6676..6681,6682..6699,6700..6706)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6714..6730
                     /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)."
     primer_bind     6719..6741
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6738..6754
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6738..6754
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6773..6793
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6775..6793
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6821..7047
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7048..7228
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7275..7400
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7893..8126
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8311..8355
                     /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"
     promoter        8533..8781
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8533..8553
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8704..8723
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        10663..10738
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    10794..10911
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
      421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1261 cgtgacgtac ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     1321 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     1381 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     1441 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg
     1501 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     1561 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     1621 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
     1681 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     1741 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     1801 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     1861 gcaagcccgg tgccggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
     1921 tcgaagagaa tcctggaccg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1981 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     2041 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     2101 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     2161 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     2221 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     2281 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     2341 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     2401 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     2461 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     2521 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     2581 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2641 acgagctgta caagtaaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg
     2701 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2761 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2821 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     2881 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2941 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3001 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3061 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3121 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3181 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3241 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
     3301 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     3361 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     3421 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     3481 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     3541 ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca
     3601 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     3661 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3721 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     3781 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3841 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3901 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     3961 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     4021 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4081 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4141 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4201 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4261 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4321 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4381 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4441 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4501 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4561 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     4621 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4681 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4741 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4801 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4861 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4921 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4981 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5041 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5101 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5161 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5221 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5281 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5341 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5401 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5461 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5521 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5581 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5641 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5701 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5761 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5821 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5881 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5941 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6001 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6061 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6121 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6181 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6241 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6301 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6361 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6421 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6481 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     6541 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     6601 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     6661 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     6721 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     6781 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     6841 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     6901 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     6961 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     7021 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     7081 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7141 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7201 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     7261 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     7321 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     7381 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     7441 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     7501 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     7561 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     7621 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     7681 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     7741 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     7801 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     7861 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     7921 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     7981 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     8041 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     8101 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     8161 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     8221 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     8281 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     8341 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     8401 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     8461 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     8521 aggggaccca gagagggcct atttcccatg attccttcat atttgcatat acgatacaag
     8581 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
     8641 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     8701 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     8761 ttgtggaaag gacgaaacac cggagacggt tgtaaatgag cacacaaaat acacatgcta
     8821 aaatattata ttctatgacc tttataaaat caaccaaaat cttcttttta ataactttag
     8881 tatcaataat tagaattttt atgttccttt ttgcaaactt ttaataaaaa tgagcaaaat
     8941 aaaaaaacgc tagttttagt aactcgcgtt gttttcttca cctttaataa tagctactcc
     9001 accacttgtt cctaagcggt cagctcctgc ttcaatcatt ttttgagcat cttcaaatgt
     9061 tctaactcca ccagctgctt taactaaagc attgtcttta acaactgact tcattagttt
     9121 aacatcttca aatgttgcac ctgattttga aaatcctgtt gatgttttaa caaattctaa
     9181 tccagcttca acagctattt cacaagcttt catgatttct tcttttgtta ataaacaatt
     9241 ttccataata catttaacaa catgtgatcc agctgctttt tttacagctt tcatgtcttc
     9301 taaaactaat tcataatttt tgtcttttaa tgcaccaata tttaatacca tatcaatttc
     9361 tgttgcacca tctttaattg cttcagaaac ttcgaatgct tttgtagctg ttgtgcatgc
     9421 acctagagga aaacctacaa catttgttat tcctacattt gtgcctttta ataattcttt
     9481 acaatagctt gttcaatatg aattaacaca aactgttgca aaatcaaatt caattgcttc
     9541 atcacataat tgtttaattt cagctttcgt agcatcttgt tttaataatg tgtgatctat
     9601 atatttgttt agtttcattt tttctcctat atattcattt ttaattttaa ttctttaata
     9661 atttcgtcta ctttaacttt agcgttttga acagattcac caacacctat aaaataaatt
     9721 tttagtttag gttcagttcc acttgggcga acagcaaatc atgacttatc ttctaaataa
     9781 aattttagta agtcttgtcc tggcatatta tacattccat cgatgtagtc ttcaacatta
     9841 acaactttaa gtccagcaat ttgagttaag ggtgttgctc tcaatgattt cattaatggt
     9901 tcaattttta atttcttttc ttctggttta aaattcaagt ttaaagtgaa agtgtaatat
     9961 gcacccattt ctttaaataa atcttctaaa tagtctacta atgttttatt ttgtttttta
    10021 taaaatcaag cagcctctgc tattaatata gaagcttgta ttccatcttt atctctagct
    10081 gagtcatcaa ttacatatcc ataactttct tcataagcaa aaacaaaatt taatccgtta
    10141 tcttcttctt tagcaatttc tctacccatt catttaaatc cagttaaagt ttttacaata
    10201 ttaactccat atttttcatg agcgattcta tcacccaaat cacttgttac aaaacttgaa
    10261 tatagagccg gattttttgg aatgctattt aagcgtttta gatttgataa ttttcaatca
    10321 attaaaattg gtcctgtttg atttccatct aatcttacaa aatgaccatc atgttttatt
    10381 gccattccaa atctgtcagc atctgggtca ttcataataa taatatctgc atcatgttta
    10441 ataccatatt caagcggtat ttttcatgca ggatcaaatt ctggatttgg atttacaaca
    10501 tttttaaatg tttcatcttc aaatgcatgc tcttcaacct caataacgtt atatcctgat
    10561 tcacgtaata tttttggggt aaatttagtt cctgttccat taactgcgct aaaaataatt
    10621 tttaaatctt ttttagcttc ttgctctttt ttgtacgtct ctgttttaga gctagaaata
    10681 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
    10741 ttttaagctt ggcgtaacta gatcttgaga caaatggcag tattcatcca caattttaaa
    10801 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
    10861 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
    10921 tacagggaca gcagagatcc actttggcgc cggctcgagg ggg
//
