LOCUS       sequence-481788-       13976 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  For lentiviral expression of PEmax with EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13976)
  AUTHORS   Niu X, Tang W, Liu Y, Liu Y, Mo B, Yu Y, Wei W
  TITLE     Prime editor-based high-throughput screening reveals functional 
            synonymous mutations in human cells
  JOURNAL   bioRxiv 2024.06.16.599253
REFERENCE   2  (bases 1 to 13976)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.06.16.599253"
COMMENT     Sequence Label: pLenti-CMV-PEmax-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..13976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1802..1918
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        1936..2315
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2316..2519
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2470..2490
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2561..2579
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2629..2649
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2650..6750
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6808..6828
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8929..8949
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8959..8985
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             8995..9051
                     /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(9052..9054,9055..9057,9058..9768)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(9052..9054,9055..9057,9058..9768)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9358..9377)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9705..9726
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             9781..9801
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    9820..10408
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9873..9893)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10291..10302)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10495..10728
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10800..10921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10837..10856)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10891..10910
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10961..11096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(11117..11135)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11145..11162)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11145..11161)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11154..11176)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11303..11758
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11390..11409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11600..11621
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11784..11888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11889..11957,11958..12749)
                     /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(12107..12126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12920..13508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13409..13428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13662..13679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13796..13817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13832..13837,13838..13855,13856..13862)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13870..13886
                     /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     13875..13897
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13894..13910
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13894..13910
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13929..13949
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13931..13949
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat
     1921 cgatggcgac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     1981 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     2041 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     2101 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     2161 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     2221 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     2281 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     2341 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     2401 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     2461 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     2521 gtttagtgaa ccgtcagatc cgctagagat ccgcggccgc taatacgact cactataggg
     2581 agagccgcca ccatgaaacg gacagccgac ggaagcgagt tcgagtcacc aaagaagaag
     2641 cggaaagtcg acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg
     2701 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     2761 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
     2821 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     2881 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     2941 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     3001 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     3061 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     3121 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     3181 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     3241 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     3301 agcagaaagc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     3361 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     3421 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     3481 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     3541 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     3601 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     3661 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     3721 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     3781 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaagag agaggacctg
     3841 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     3901 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     3961 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     4021 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     4081 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     4141 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     4201 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     4261 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     4321 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     4381 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     4441 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     4501 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     4561 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     4621 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     4681 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     4741 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     4801 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     4861 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     4921 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     4981 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     5041 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     5101 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     5161 gacgctatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     5221 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     5281 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     5341 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     5401 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     5461 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     5521 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     5581 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     5641 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     5701 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     5761 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     5821 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     5881 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     5941 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     6001 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     6061 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     6121 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     6181 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     6241 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     6301 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     6361 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     6421 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     6481 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     6541 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     6601 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     6661 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     6721 cggatcgacc tgtctcagct gggaggtgac tccggcggaa gctctggtgg cagcaagcgg
     6781 accgccgacg gctctgaatt cgagagccct aagaagaaaa gaaaggtgag cggaggctct
     6841 agcggcggaa gcaccctgaa cattgaagac gagtatagac tgcatgaaac aagcaaggaa
     6901 cccgacgtgt ccctgggctc cacctggctg tccgactttc cccaggcctg ggccgagaca
     6961 ggaggaatgg gcctggccgt gcggcaggca cccctgatca tccctctgaa ggccacctct
     7021 acacccgtga gcatcaagca gtaccctatg tctcaggagg ccagactggg catcaagcct
     7081 cacatccaga ggctgctgga ccagggcatc ctggtgccat gccagagccc ctggaacaca
     7141 ccactgctgc ccgtgaagaa gccaggcacc aatgactata gacccgtgca ggatctgaga
     7201 gaggtgaaca agagggtgga ggatatccac cccaccgtgc ccaaccctta caatctgctg
     7261 tccggcctgc ccccttctca ccagtggtat acagtgctgg acctgaagga tgccttcttt
     7321 tgtctgagac tgcaccctac cagccagcca ctgttcgcct ttgagtggag ggaccctgag
     7381 atgggcatct ctggccagct gacctggaca cgcctgcctc agggcttcaa gaatagccca
     7441 acactgttta acgaggccct gcaccgcgac ctggcagatt tccggatcca gcacccagat
     7501 ctgatcctgc tgcagtacgt ggacgatctg ctgctggccg ccaccagcga gctggattgc
     7561 cagcagggaa cacgcgccct gctgcagacc ctgggaaacc tgggatatag ggcatccgcc
     7621 aagaaggccc agatctgtca gaagcaggtg aagtacctgg gctatctgct gaaggagggc
     7681 cagagatggc tgacagaggc caggaaggag acagtgatgg gccagccaac acccaagacc
     7741 ccaagacagc tgagggagtt cctgggcaaa gcaggatttt gcaggctgtt catcccagga
     7801 ttcgcagaga tggcagcacc tctgtaccca ctgaccaagc cgggcaccct gtttaattgg
     7861 ggccctgacc agcagaaggc ctatcaggag atcaagcagg ccctgctgac agcaccagcc
     7921 ctgggcctgc cagacctgac caagcctttc gagctgtttg tggatgagaa gcagggctac
     7981 gccaagggcg tgctgaccca gaagctggga ccatggagac ggcccgtggc ctatctgtcc
     8041 aagaagctgg acccagtggc agcaggatgg ccaccatgcc tgaggatggt ggcagcaatc
     8101 gccgtgctga caaaggatgc cggcaagctg accatgggac agccactggt catcctggca
     8161 ccacacgcag tggaggccct ggtgaagcag cctccagatc gctggctgtc taacgcccgg
     8221 atgacacact accaggccct gctgctggac accgatcgcg tgcagtttgg ccctgtggtg
     8281 gccctgaatc cagccaccct gctgcctctg ccagaggagg gcctgcagca caactgtctg
     8341 gacatcctgg cagaggcaca cggaacaagg ccagacctga ccgatcagcc cctgcctgac
     8401 gccgatcaca catggtatac cgatggaagc tccctgctgc aggagggcca gaggaaggca
     8461 ggagcagcag tgaccacaga gacagaagtg atctgggcca aggccctgcc agcaggcaca
     8521 tccgcccagc gggccgagct gatcgccctg acccaggccc tgaagatggc cgagggcaag
     8581 aagctgaacg tgtacacaga ctccagatat gccttcgcca ccgcacacat ccacggagag
     8641 atctacaggc gccggggctg gctgacctct gagggcaagg agatcaagaa caaggatgag
     8701 atcctggccc tgctgaaggc cctgtttctg cccaagcggc tgagcatcat ccactgtcct
     8761 ggacaccaga agggacactc cgccgaggca aggggcaatc ggatggccga ccaggccgcc
     8821 agaaaggctg ctattactga aactcccgac acttccactc tgctgattga aaactcctcc
     8881 ccttctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagtctcc caagaagaag
     8941 aggaaagtcg gctctggccc tgccgctaag agagtgaagc tggacggatc cggcgctact
     9001 aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc tatggtgagc
     9061 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     9121 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     9181 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     9241 accctgacct atggagtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     9301 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     9361 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     9421 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     9481 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     9541 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     9601 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     9661 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     9721 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc tggtggttct
     9781 cccaagaaga agaggaaagt ctaaccggtg ggcgtcgaca atcaacctct ggattacaaa
     9841 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     9901 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     9961 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
    10021 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
    10081 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
    10141 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
    10201 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
    10261 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
    10321 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
    10381 cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacct ttaagaccaa
    10441 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
    10501 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
    10561 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
    10621 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
    10681 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
    10741 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
    10801 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    10861 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    10921 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
    10981 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
    11041 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
    11101 gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
    11161 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
    11221 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
    11281 cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
    11341 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
    11401 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
    11461 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
    11521 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
    11581 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
    11641 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
    11701 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
    11761 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
    11821 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
    11881 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
    11941 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
    12001 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
    12061 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
    12121 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
    12181 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
    12241 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
    12301 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
    12361 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
    12421 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
    12481 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
    12541 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
    12601 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
    12661 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
    12721 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
    12781 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
    12841 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    12901 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
    12961 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
    13021 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
    13081 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
    13141 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
    13201 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
    13261 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
    13321 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
    13381 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
    13441 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
    13501 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
    13561 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
    13621 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
    13681 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
    13741 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
    13801 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
    13861 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
    13921 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaa
//
