LOCUS       sequence-366971-       11493 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral backbone with EBFP2 reporter and gRNA targeting MFAP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11493)
  AUTHORS   Werder RB, Liu T, Abo KM, Lindstrom-Vautrin J, Villacorta-Martin C, 
            Huang J, Hinds A, Boyer N, Bullitt E, Liesa M, Silverman EK, Kotton 
            DN, Cho MH, Zhou X, Wilson AA
  TITLE     CRISPR interference interrogation of COPD GWAS genes reveals the 
            functional significance of desmoplakin in iPSC-derived alveolar 
            epithelial cells.
  JOURNAL   Sci Adv. 2022 Jul 15;8(28):eabo6566. doi: 10.1126/sciadv.abo6566. 
            Epub 2022 Jul 13.
  PUBMED    35857525
REFERENCE   2  (bases 1 to 11493)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2022-07-15"; volume: "8(28):eabo6566. doi"; pages: " 
            10.1126/sciadv.abo6566. Epub 2022 Jul 13"
COMMENT     Sequence Label: pLV-GG-hUbC-EBPF2-gRNA-MFAP2
FEATURES             Location/Qualifiers
     source          1..11493
                     /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"
     protein_bind    complement(2602..2635)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2640..2953
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2872..2892
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        2976..3051
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(3081..3156)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(3178..3426)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(3236..3255)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(3406..3426)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        3456..3670
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     3591..3610
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        3700..3775
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(3805..3880)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4177..5388
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5225..5244
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      5435..5444
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5441..5443,5444..5446,5447..6160)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(5486..5507)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    6185..6773
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6238..6258)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6656..6667)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6776..6792)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6777..6793)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7298..7478
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7504..7521)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7510..7734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7780..8208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7867..7886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8077..8098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8217..8237)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8222..8551
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8402..8537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8464..8483
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8599..8646
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8665..9039
                     /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    9169..9290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9206..9225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9260..9279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9339..9355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9339..9355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9352..9374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9363..9379
                     /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(9387..9393,9394..9411,9412..9417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9432..9453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9570..9587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9741..10329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9821..9840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10500..11291,11292..11360))
                     /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     11123..11142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11361..11465)
                     /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 aataacttcg tatagcatac attatacgaa gttatgatag
     2641 atccgacgcc gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc
     2701 tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct
     2761 taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat
     2821 aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa acagcacaaa
     2881 aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga
     2941 gaaaagcctt gtttgactta ccaggcagga atagcgtttt agagctagaa atagcaagtt
     3001 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgt
     3061 acgctcagcg gagaaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact
     3121 agccttattt taacttgcta tttctagctc taaaacacat aagaggagct accccccggt
     3181 gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc aagttacggt
     3241 aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc caagaaatta
     3301 ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca aattaattcc
     3361 aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag
     3421 gccctcttcc tgcccgacct tggatcgatc cgcctgaacg ctgacgtcat caacccgctc
     3481 caaggaatcg cgggcccagt gtcactaggc gggaacaccc agcgcgcgtg cgccctggca
     3541 ggaagatggc tgtgagggac aggggagtgg cgccctgcaa tatttgcatg tcgctatgtg
     3601 ttctgggaaa tcaccataaa cgtgaaatgt ctttggattt gggaatctta taagttctgt
     3661 atgagaccac tctttcccat tgtcacgact tatgaccagg ttttagagct agaaatagca
     3721 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     3781 ttggtcatca gtccgagaaa aaaagcaccg actcggtgcc actttttcaa gttgataacg
     3841 gactagcctt attttaactt gctatttcta gctctaaaac ctactaaccc tactcctcag
     3901 cgaggtaccc aagcggcgca caagctatat aaacctgaag gaagtctcaa ctttacactt
     3961 aggtcaagtt gcttatcgta ctagagcttc agcaggaaat ttaactaaaa tctaatttaa
     4021 ccagcatagc aaatatcatt tattcccaaa atgctaaagt ttgagataaa cggacttgat
     4081 ttccggctgt tttgacacta tccagaatgc cttgcagatg ggtggggcat gctaaatact
     4141 gcagacaatt aattaacccg tgtcggctcc agatctggcc tccgcgccgg gttttggcgc
     4201 ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg
     4261 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     4321 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     4381 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     4441 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     4501 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     4561 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     4621 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     4681 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     4741 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     4801 atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt
     4861 cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     4921 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     4981 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     5041 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     5101 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     5161 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     5221 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     5281 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     5341 agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga agcttgggct
     5401 gcaggtcgac tctagaggat ccccgggtac cggtgccacc atggtgagca agggcgagga
     5461 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     5521 gttcagcgtg aggggcgagg gcgagggcga tgccaccaac ggcaagctga ccctgaagtt
     5581 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgagcca
     5641 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     5701 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaccta
     5761 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     5821 gggcgtcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacttcaa
     5881 cagccacaac atctatatca tggccgtcaa gcagaagaac ggcatcaagg tgaacttcaa
     5941 gatccgccac aacgtggagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     6001 ccccatcggc gacggccccg tgctgctgcc cgacagccac tacctgagca cccagtccgt
     6061 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tccgcaccgc
     6121 cgccgggatc actctcggca tggacgagct gtacaagtaa gaattcgata tcaagcttat
     6181 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     6241 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     6301 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     6361 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     6421 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     6481 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     6541 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     6601 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6661 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     6721 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     6781 ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     6841 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
     6901 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     6961 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     7021 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc
     7081 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga
     7141 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca
     7201 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt
     7261 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat
     7321 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7381 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7441 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
     7501 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     7561 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     7621 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     7681 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     7741 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     7801 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     7861 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     7921 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     7981 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8041 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     8101 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     8161 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     8221 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     8281 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     8341 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     8401 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     8461 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     8521 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     8581 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     8641 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     8701 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     8761 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     8821 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     8881 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     8941 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     9001 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     9061 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     9121 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     9181 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     9241 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     9301 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     9361 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     9421 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     9481 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     9541 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     9601 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     9661 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     9721 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     9781 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     9841 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     9901 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     9961 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
    10021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
    10081 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
    10141 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
    10201 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
    10261 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
    10321 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
    10381 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
    10441 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
    10501 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    10561 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    10621 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    10681 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    10741 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    10801 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    10861 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    10921 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    10981 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    11041 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    11101 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    11161 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    11221 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    11281 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    11341 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    11401 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    11461 ccgcgcacat ttccccgaaa agtgccacct gac
//
