LOCUS       sequence-271340-       12226 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentivirus for constitutive expression of FNLSHiFiNG in mammalian 
            cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12226)
  AUTHORS   Katti A, Foronda M, Zimmerman J, Diaz B, Zafra MP, Goswami S, Dow LE
  TITLE     GO: a functional reporter system to identify and enrich base editing
            activity.
  JOURNAL   Nucleic Acids Res. 2020 Feb 29. pii: 5766653. doi: 
            10.1093/nar/gkaa124.
  PUBMED    32112097
REFERENCE   2  (bases 1 to 12226)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res. 2020 Feb 29. pii: 5766653. doi: 10.1093/nar/gkaa124."
COMMENT     Sequence Label: pLenti-FNLSHiFiNG-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..12226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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    1723..1840
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1909..2120
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2140..2149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2149..2214
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2221..2241
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2266..2952
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             3001..7101
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANANFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7114..7362
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7375..7395
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7411..7467
                     /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             7468..8067
                     /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(7468..7487)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7964..7984
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8083..8671
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8136..8156)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8554..8565)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8743..8976
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9054..9175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9091..9110)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9145..9164
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9215..9350
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9370..9389)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9371..9389)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9399..9416)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9399..9415)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9408..9430)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9557..10012
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9644..9663)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9854..9875
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10038..10142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10143..10211,10212..11003)
                     /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(10361..10380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11174..11762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11663..11682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11916..11933
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12050..12071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12086..12091,12092..12109,12110..12116)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12124..12140
                     /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     12129..12151
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12148..12164
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12148..12164
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12183..12203
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12185..12203
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg
     1741 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1801 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1861 cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca
     1921 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1981 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2041 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2101 ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga ttacaaagat
     2161 catgacggcg actataagga ccacgatatt gactacaaag atgatgacga caagatggct
     2221 cccaaaaaga aaaggaaagt aggcattcat ggagttcccg cagccatgag ttctgagact
     2281 ggtccagtcg ccgtcgatcc aacccttcga cggaggattg agccccacga gtttgaagta
     2341 tttttcgatc caagagaatt gcgaaaagag acatgtcttc tgtacgagat caactggggg
     2401 ggccgccaca gtatttggag acacaccagc cagaacacaa ataagcatgt cgaggtaaat
     2461 ttcatagaga agttcactac cgaacgctac ttttgcccta acacaagatg tagcattact
     2521 tggttcttga gttggtcccc ttgcggtgaa tgctcaagag caattaccga gtttctgtct
     2581 cggtatcccc acgtcacttt gttcatctac atagcccgac tttaccatca cgcagaccct
     2641 cgaaatcgcc agggtcttag agatttgatt tcatccggtg taaccataca aattatgacc
     2701 gaacaagaga gtgggtattg ttggcggaat ttcgtcaact acagtccatc aaatgaggcc
     2761 cattggcccc gctaccccca cctttgggtg agattgtacg tattggagtt gtattgcata
     2821 atcctgggcc ttcccccctg tcttaacatc cttagaagga agcagcccca gttgacattc
     2881 tttactatcg ctcttcagtc atgtcactat cagcgactgc ctcctcatat cctttgggct
     2941 actgggttga aaagcggttc cgagacacct ggtacttcag agtccgccac acccgaaagt
     3001 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
     3061 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     3121 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     3181 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     3241 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     3301 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     3361 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     3421 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     3481 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     3541 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     3601 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     3661 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg
     3721 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     3781 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     3841 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     3901 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     3961 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     4021 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     4081 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     4141 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     4201 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     4261 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     4321 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     4381 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     4441 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     4501 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     4561 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     4621 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     4681 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     4741 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     4801 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     4861 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     4921 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     4981 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     5041 ttcctgaagt ccgacggctt cgccaacgcc aacttcatgc agctgatcca cgacgacagc
     5101 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     5161 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     5221 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     5281 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     5341 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     5401 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     5461 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     5521 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     5581 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     5641 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     5701 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     5761 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     5821 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     5881 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     5941 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     6001 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     6061 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     6121 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     6181 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     6241 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     6301 acaggcggct tcagcaaaga gtctatcagg cccaagagga acagcgataa gctgatcgcc
     6361 agaaagaagg actgggaccc taagaagtac ggcggcttcg tcagccccac cgtggcctat
     6421 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     6481 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     6541 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     6601 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccag gttcctgcag
     6661 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     6721 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     6781 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     6841 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     6901 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     6961 cgggccttca agtactttga caccaccatc gaccggaagg tgtacaggag caccaaagag
     7021 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     7081 ctgtctcagc tgggaggcga ttcaggcgga tctactaatc tgtcagatat tattgaaaag
     7141 gagaccggta agcaactggt tatccaggaa tccatcctca tgctcccaga ggaggtggaa
     7201 gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc acaccgccta cgacgagagc
     7261 accgacgaga atgtcatgct tctgactagc gacgcccctg aatacaagcc ttgggctctg
     7321 gtcatacagg atagcaacgg tgagaacaag attaagatgc tctctggtgg ttctcccaag
     7381 aagaagagga aagtcgctag cggcagcggc gccaccaact tcagcctgct gaagcaggcc
     7441 ggcgacgtgg aggagaaccc cggccccatg accgagtaca agcccacggt gcgcctcgcc
     7501 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     7561 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     7621 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     7681 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     7741 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     7801 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg
     7861 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     7921 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     7981 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     8041 tgcatgaccc gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac
     8101 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     8161 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     8221 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     8281 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     8341 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     8401 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     8461 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     8521 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     8581 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     8641 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     8701 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     8761 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     8821 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     8881 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     8941 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     9001 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     9061 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     9121 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     9181 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     9241 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     9301 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     9361 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     9421 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     9481 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     9541 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     9601 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     9661 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     9721 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     9781 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     9841 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     9901 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     9961 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
    10021 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
    10081 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
    10141 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
    10201 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
    10261 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
    10321 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
    10381 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
    10441 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
    10501 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
    10561 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
    10621 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
    10681 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
    10741 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
    10801 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
    10861 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
    10921 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
    10981 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
    11041 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
    11101 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
    11161 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
    11221 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
    11281 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
    11341 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
    11401 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
    11461 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
    11521 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
    11581 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    11641 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    11701 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    11761 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    11821 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
    11881 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
    11941 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
    12001 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    12061 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    12121 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
    12181 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagc
//
