LOCUS       sequence-266659-        9651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP in mammalian cells, contains a 3' MCS. Localized to 
            the nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9651)
  AUTHORS   Wessels HH, Mendez-Mancilla A, Guo X, Legut M, Daniloski Z, Sanjana 
            NE
  TITLE     Massively parallel Cas13 screens reveal principles for guide RNA 
            design.
  JOURNAL   Nat Biotechnol. 2020 Jun;38(6):722-727. doi: 
            10.1038/s41587-020-0456-9. Epub 2020 Mar 16.
  PUBMED    32518401
REFERENCE   2  (bases 1 to 9651)
  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"; doi: 
            "10.1038/s41587-020-0456-9"; journalName: "Nat Biotechnol"; date: 
            "2020-06"; volume: "38"; issue: "6"; pages: "722-727"
COMMENT     Sequence Label: pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-...
FEATURES             Location/Qualifiers
     source          1..9651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..1274
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          327..1265
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1222..1242
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1288..1297
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1294..1296,1297..1299,1300..2010,2011..2016,
                     2017..2136)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(1339..1360)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1600..1619)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1947..1968
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2137..2184
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     misc_feature    2249..2837
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2302..2322)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2720..2731)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2909..3142
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3220..3341
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3257..3276)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3311..3330
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3381..3516
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3536..3555)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3537..3555)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3565..3582)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3565..3581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3574..3596)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3723..4178
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3810..3829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4020..4041
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4204..4308
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4309..4377,4378..5169)
                     /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(4527..4546)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5340..5928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5829..5848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6082..6099
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6216..6237
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6252..6257,6258..6275,6276..6282)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6290..6306
                     /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     6295..6317
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6314..6330
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6314..6330
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6349..6369
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6351..6369
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6397..6623
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6624..6804
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6851..6976
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7469..7702
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7887..7931
                     /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        8162..8372
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        8184..8380
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8231..8366
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8402..9421
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     misc_feature    9492..9609
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctcgaggcct gcaggtgcaa agatggataa agttttaaac agagaggaat ctttgcagct
       61 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
      121 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
      181 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      241 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      301 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
      361 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
      421 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
      481 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
      541 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
      601 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
      661 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
      721 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
      781 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
      841 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
      901 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
      961 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1021 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1081 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1141 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1201 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1261 ttcaggtgtc gtgaggctag catcgatgcc accatggtga gcaagggcga ggagctgttc
     1321 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1381 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1441 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1501 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1561 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1621 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1681 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1741 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1801 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1861 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1921 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1981 atcactctcg gcatggacga gctgtacaag aagcttagcc atggcttccc gccggaggtg
     2041 gaggagcagg atgatggcac gctgcccatg tcttgtgccc aggagagcgg gatggaccgt
     2101 caccctgcag cctgtgcttc tgctaggatc aatgtgaagc gacctgccgc cacaaagaag
     2161 gctggacagg ctaagaagaa gaaatgagga tcccgcgcgc gcatatgtta attaaccaac
     2221 tgcatgggga tccacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     2281 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2341 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2401 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2461 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2521 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2581 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2641 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2701 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2761 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2821 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     2881 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     2941 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     3001 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     3061 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     3121 tcagtgtgga aaatctctag cagtacgtat agtagttcat gtcatcttat tattcagtat
     3181 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     3241 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3301 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     3361 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3421 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3481 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     3541 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     3601 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     3661 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3721 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3781 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3841 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3901 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3961 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4021 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4081 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4141 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4201 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4261 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4321 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4381 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4441 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4501 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4561 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4621 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4681 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4741 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4801 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4861 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4921 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4981 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5041 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5101 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5161 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5221 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5281 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5341 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5401 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5461 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5521 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5581 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5641 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5701 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5761 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5821 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5881 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5941 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6001 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6061 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6121 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6181 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6241 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6301 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
     6361 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt
     6421 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
     6481 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
     6541 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
     6601 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
     6661 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6721 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6781 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
     6841 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     6901 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     6961 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
     7021 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
     7081 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
     7141 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
     7201 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
     7261 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     7321 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     7381 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     7441 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     7501 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     7561 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     7621 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     7681 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     7741 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     7801 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     7861 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     7921 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     7981 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     8041 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     8101 gacccatgca ttgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     8161 agcaggcaga agtatgcaaa gcatgcgtct caattagtca gcaaccatag tcccgcccct
     8221 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     8281 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     8341 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctttcta gaggtaccac
     8401 catgaaaaag cctgaactca ccgctacctc tgtcgagaag tttctgatcg aaaagttcga
     8461 cagcgtctcc gacctgatgc agctctccga gggcgaagaa tctcgggctt tcagcttcga
     8521 tgtgggaggg cgtggatatg tcctgcgggt gaatagctgc gccgatggtt tctacaaaga
     8581 tcgctatgtt tatcggcact ttgcatccgc cgctctccct attcccgaag tgcttgacat
     8641 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacctt
     8701 gcaagacctg cctgaaaccg aactgcccgc tgttctccag cccgtcgccg aggccatgga
     8761 tgccatcgct gccgccgatc ttagccagac cagcgggttc ggcccattcg gacctcaagg
     8821 aatcggtcaa tacactacat ggcgcgattt catctgcgct attgctgatc cccatgtgta
     8881 tcactggcaa actgtgatgg acgacaccgt cagtgcctcc gtcgcccagg ctctcgatga
     8941 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg ccgatttcgg
     9001 ctccaacaat gtcctgaccg acaatggccg cataacagcc gtcattgact ggagcgaggc
     9061 catgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cctggttggc
     9121 ttgtatggag cagcagaccc gctacttcga gcggaggcat cccgagcttg caggatctcc
     9181 tcggctccgg gcttatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     9241 cggcaatttc gatgatgcag cttgggctca gggtcgctgc gacgcaatcg tccggtccgg
     9301 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gctgccgtct ggaccgatgg
     9361 ctgtgtggaa gtgctcgccg atagtggaaa cagacgcccc agcactcgtc ctagggcaaa
     9421 ggatctgcag taatgaggcg cgccgttaac gaattctaga tcttgagaca aatggcagta
     9481 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     9541 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     9601 caaaattttc gggtttatta cagggacagc agagatccac tttggcgccg g
//
