LOCUS       sequence-454001-       13372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral delivery and expression of PEmax protein with puromycin 
            selection cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13372)
  AUTHORS   Gould SI, Wuest AN, Dong K, Johnson GA, Hsu A, Narendra VK, Atwa O, 
            Levine SS, Liu DR, Sanchez Rivera FJ
  TITLE     High-throughput evaluation of genetic variants with prime editing 
            sensor libraries.
  JOURNAL   Nat Biotechnol. 2024 Mar 12. doi: 10.1038/s41587-024-02172-9.
  PUBMED    38472508
REFERENCE   2  (bases 1 to 13372)
  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: "Nat 
            Biotechnol. 2024 Mar 12. doi: 10.1038/s41587-024-02172-9."
COMMENT     Sequence Label: Lenti-EFS-PEmax-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..13372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2130..2363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2548..2592
                     /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    2776..2893
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2962..3173
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3235..3255
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3256..7356
                     /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             7414..7434
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9535..9555
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9565..9591
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             9610..9666
                     /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             9667..10266
                     /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(9667..9686)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10163..10183
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10282..10870
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10335..10355)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10753..10764)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10942..11175
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    11253..11374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11290..11309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11344..11363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      11414..11549
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(11569..11588)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(11570..11588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11598..11615)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11598..11614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11607..11629)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      11756..12211
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11843..11862)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12053..12074
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        12237..12341
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12342..12410,12411..13202)
                     /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(12560..12579)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttattccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2761 ggacccagac cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     2821 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     2881 aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga
     2941 gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     3001 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     3061 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     3121 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtgtcgt
     3181 gacgcgggat ccgccaccat gaaacggaca gccgacggaa gcgagttcga gtcaccaaag
     3241 aagaagcgga aagtcgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
     3301 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     3361 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     3421 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     3481 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     3541 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     3601 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     3661 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     3721 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     3781 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     3841 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     3901 agcaagagca gaaagctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     3961 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     4021 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     4081 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     4141 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     4201 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     4261 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     4321 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     4381 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaagagagag
     4441 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     4501 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     4561 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     4621 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     4681 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     4741 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     4801 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     4861 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     4921 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     4981 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     5041 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     5101 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     5161 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     5221 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     5281 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     5341 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     5401 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     5461 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     5521 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     5581 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     5641 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     5701 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     5761 gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     5821 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     5881 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     5941 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     6001 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     6061 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     6121 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     6181 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     6241 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     6301 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     6361 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     6421 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     6481 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     6541 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     6601 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     6661 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     6721 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     6781 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     6841 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     6901 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     6961 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     7021 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     7081 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     7141 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     7201 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     7261 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     7321 gagacacgga tcgacctgtc tcagctggga ggtgactccg gcggaagctc tggtggcagc
     7381 aagcggaccg ccgacggctc tgaattcgag agccctaaga agaaaagaaa ggtgagcgga
     7441 ggctctagcg gcggaagcac cctgaacatt gaagacgagt atagactgca tgaaacaagc
     7501 aaggaacccg acgtgtccct gggctccacc tggctgtccg actttcccca ggcctgggcc
     7561 gagacaggag gaatgggcct ggccgtgcgg caggcacccc tgatcatccc tctgaaggcc
     7621 acctctacac ccgtgagcat caagcagtac cctatgtctc aggaggccag actgggcatc
     7681 aagcctcaca tccagaggct gctggaccag ggcatcctgg tgccatgcca gagcccctgg
     7741 aacacaccac tgctgcccgt gaagaagcca ggcaccaatg actatagacc cgtgcaggat
     7801 ctgagagagg tgaacaagag ggtggaggat atccacccca ccgtgcccaa cccttacaat
     7861 ctgctgtccg gcctgccccc ttctcaccag tggtatacag tgctggacct gaaggatgcc
     7921 ttcttttgtc tgagactgca ccctaccagc cagccactgt tcgcctttga gtggagggac
     7981 cctgagatgg gcatctctgg ccagctgacc tggacacgcc tgcctcaggg cttcaagaat
     8041 agcccaacac tgtttaacga ggccctgcac cgcgacctgg cagatttccg gatccagcac
     8101 ccagatctga tcctgctgca gtacgtggac gatctgctgc tggccgccac cagcgagctg
     8161 gattgccagc agggaacacg cgccctgctg cagaccctgg gaaacctggg atatagggca
     8221 tccgccaaga aggcccagat ctgtcagaag caggtgaagt acctgggcta tctgctgaag
     8281 gagggccaga gatggctgac agaggccagg aaggagacag tgatgggcca gccaacaccc
     8341 aagaccccaa gacagctgag ggagttcctg ggcaaagcag gattttgcag gctgttcatc
     8401 ccaggattcg cagagatggc agcacctctg tacccactga ccaagccggg caccctgttt
     8461 aattggggcc ctgaccagca gaaggcctat caggagatca agcaggccct gctgacagca
     8521 ccagccctgg gcctgccaga cctgaccaag cctttcgagc tgtttgtgga tgagaagcag
     8581 ggctacgcca agggcgtgct gacccagaag ctgggaccat ggagacggcc cgtggcctat
     8641 ctgtccaaga agctggaccc agtggcagca ggatggccac catgcctgag gatggtggca
     8701 gcaatcgccg tgctgacaaa ggatgccggc aagctgacca tgggacagcc actggtcatc
     8761 ctggcaccac acgcagtgga ggccctggtg aagcagcctc cagatcgctg gctgtctaac
     8821 gcccggatga cacactacca ggccctgctg ctggacaccg atcgcgtgca gtttggccct
     8881 gtggtggccc tgaatccagc caccctgctg cctctgccag aggagggcct gcagcacaac
     8941 tgtctggaca tcctggcaga ggcacacgga acaaggccag acctgaccga tcagcccctg
     9001 cctgacgccg atcacacatg gtataccgat ggaagctccc tgctgcagga gggccagagg
     9061 aaggcaggag cagcagtgac cacagagaca gaagtgatct gggccaaggc cctgccagca
     9121 ggcacatccg cccagcgggc cgagctgatc gccctgaccc aggccctgaa gatggccgag
     9181 ggcaagaagc tgaacgtgta cacagactcc agatatgcct tcgccaccgc acacatccac
     9241 ggagagatct acaggcgccg gggctggctg acctctgagg gcaaggagat caagaacaag
     9301 gatgagatcc tggccctgct gaaggccctg tttctgccca agcggctgag catcatccac
     9361 tgtcctggac accagaaggg acactccgcc gaggcaaggg gcaatcggat ggccgaccag
     9421 gccgccagaa aggctgctat tactgaaact cccgacactt ccactctgct gattgaaaac
     9481 tcctcccctt ctggcggctc aaaaagaacc gccgacggca gcgaattcga gtctcccaag
     9541 aagaagagga aagtcggctc tggccctgcc gctaagagag tgaagctgga cggagctagc
     9601 ggcagcggcg ccaccaactt cagcctgctg aagcaggccg gcgacgtgga ggagaacccc
     9661 ggccccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     9721 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     9781 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     9841 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     9901 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     9961 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
    10021 aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt
    10081 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
    10141 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
    10201 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
    10261 gcctgaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact
    10321 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
    10381 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
    10441 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
    10501 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
    10561 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
    10621 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
    10681 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
    10741 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
    10801 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
    10861 gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact
    10921 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
    10981 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
    11041 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
    11101 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
    11161 ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac
    11221 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
    11281 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    11341 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
    11401 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
    11461 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
    11521 gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag
    11581 tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
    11641 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
    11701 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
    11761 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
    11821 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
    11881 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
    11941 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
    12001 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
    12061 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
    12121 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
    12181 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
    12241 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
    12301 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
    12361 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
    12421 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
    12481 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
    12541 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
    12601 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
    12661 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
    12721 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
    12781 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
    12841 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
    12901 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
    12961 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
    13021 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
    13081 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
    13141 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
    13201 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
    13261 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
    13321 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tc
//
