LOCUS       sequence-292713-       11137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression plasmid of AsCas12a-6xNLS-E174R/S542R with 
            puromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11137)
  AUTHORS   Gier RA, Budinich KA, Evitt NH, Cao Z, Freilich ES, Chen Q, Qi J, 
            Lan Y, Kohli RM, Shi J
  TITLE     High-performance CRISPR-Cas12a genome editing for combinatorial 
            genetic screening.
  JOURNAL   Nat Commun. 2020 Jul 13;11(1):3455. doi: 10.1038/s41467-020-17209-1.
  PUBMED    32661245
REFERENCE   2  (bases 1 to 11137)
  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 
            Commun."; date: "2020-07-13"; volume: "11(1):3455. doi"; pages: " 
            10.1038/s41467-020-17209-1"
COMMENT     Sequence Label: AsCas12a-6xNLS-E174R/S542R (pRG232)
FEATURES             Location/Qualifiers
     source          1..11137
                     /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..2172
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2173..6090
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      5810..5819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6100..6126
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6136..6162
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6172..6198
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6208..6234
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6244..6270
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6280..6306
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6322..6378
                     /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             6379..6978
                     /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(6379..6398)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6875..6895
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6994..7582
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7047..7067)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7465..7476)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7654..7887
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7965..8086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8002..8021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8056..8075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8126..8261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8281..8300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8282..8300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8310..8327)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8310..8326)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8319..8341)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8468..8923
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8555..8574)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8765..8786
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8949..9053
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9054..9122,9123..9914)
                     /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(9272..9291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10085..10673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10574..10593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10827..10844
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10961..10982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10997..11002,11003..11020,11021..11027)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11035..11051
                     /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     11040..11062
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11059..11075
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11059..11075
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11094..11114
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11096..11114
                     /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 ttacaaagac
     2161 gatgacgata agacacagtt cgagggcttt accaacctgt atcaggtgag caagacactg
     2221 cggtttgagc tgatcccaca gggcaagacc ctgaagcaca tccaggagca gggcttcatc
     2281 gaggaggaca aggcccgcaa tgatcactac aaggagctga agcccatcat cgatcggatc
     2341 tacaagacct atgccgacca gtgcctgcag ctggtgcagc tggattggga gaacctgagc
     2401 gccgccatcg actcctatag aaaggagaaa accgaggaga caaggaacgc cctgatcgag
     2461 gagcaggcca catatcgcaa tgccatccac gactacttca tcggccggac agacaacctg
     2521 accgatgcca tcaataagag acacgccgag atctacaagg gcctgttcaa ggccgagctg
     2581 tttaatggca aggtgctgaa gcagctgggc accgtgacca caaccgagca cgagaacgcc
     2641 ctgctgcgga gcttcgacaa gtttacaacc tacttctccg gcttttatcg gaacaggaag
     2701 aacgtgttca gcgccgagga tatcagcaca gccatcccac accgcatcgt gcaggacaac
     2761 ttccccaagt ttaaggagaa ttgtcacatc ttcacacgcc tgatcaccgc cgtgcccagc
     2821 ctgcgggagc actttgagaa cgtgaagaag gccatcggca tcttcgtgag cacctccatc
     2881 gaggaggtgt tttccttccc tttttataac cagctgctga cacagaccca gatcgacctg
     2941 tataaccagc tgctgggagg aatctctcgg gaggcaggca ccgagaagat caagggcctg
     3001 aacgaggtgc tgaatctggc catccagaag aatgatgaga cagcccacat catcgcctcc
     3061 ctgccacaca gattcatccc cctgtttaag cagatcctgt ccgataggaa caccctgtct
     3121 ttcatcctgg aggagtttaa gagcgacgag gaagtgatcc agtccttctg caagtacaag
     3181 acactgctga gaaacgagaa cgtgctggag acagccgagg ccctgtttaa cgagctgaac
     3241 agcatcgacc tgacacacat cttcatcagc cacaagaagc tggagacaat cagcagcgcc
     3301 ctgtgcgacc actgggatac actgaggaat gccctgtatg agcggagaat ctccgagctg
     3361 acaggcaaga tcaccaagtc tgccaaggag aaggtgcagc gcagcctgaa gcacgaggat
     3421 atcaacctgc aggagatcat ctctgccgca ggcaaggagc tgagcgaggc cttcaagcag
     3481 aaaaccagcg agatcctgtc ccacgcacac gccgccctgg atcagccact gcctacaacc
     3541 ctgaagaagc aggaggagaa ggagatcctg aagtctcagc tggacagcct gctgggcctg
     3601 taccacctgc tggactggtt tgccgtggat gagtccaacg aggtggaccc cgagttctct
     3661 gcccggctga ccggcatcaa gctggagatg gagccttctc tgagcttcta caacaaggcc
     3721 agaaattatg ccaccaagaa gccctactcc gtggagaagt tcaagctgaa ctttcagatg
     3781 cctacactgg cccgtggctg ggacgtgaat aaggagaaga acaatggcgc catcctgttt
     3841 gtgaagaacg gcctgtacta tctgggcatc atgccaaagc agaagggcag gtataaggcc
     3901 ctgagcttcg agcccacaga gaaaaccagc gagggctttg ataagatgta ctatgactac
     3961 ttccctgatg ccgccaagat gatcccaaag tgcagcaccc agctgaaggc cgtgacagcc
     4021 cactttcaga cccacacaac ccccatcctg ctgtccaaca atttcatcga gcctctggag
     4081 atcacaaagg agatctacga cctgaacaat cctgagaagg agccaaagaa gtttcagaca
     4141 gcctacgcca agaaaaccgg cgaccagaag ggctacagag aggccctgtg caagtggatc
     4201 gacttcacaa gggattttct gtccaagtat accaagacaa cctctatcga tctgtctagc
     4261 ctgcggccat cctctcagta taaggacctg ggcgagtact atgccgagct gaatcccctg
     4321 ctgtaccaca tcagcttcca gagaatcgcc gagaaggaga tcatggatgc cgtggagaca
     4381 ggcaagctgt acctgttcca gatctataac aaggactttg ccaagggcca ccacggcaag
     4441 cctaatctgc acacactgta ttggaccggc ctgttttctc cagagaacct ggccaagaca
     4501 agcatcaagc tgaatggcca ggccgagctg ttctaccgcc ctaagtccag gatgaagagg
     4561 atggcacacc ggctgggaga gaagatgctg aacaagaagc tgaaggatca gaaaacccca
     4621 atccccgaca ccctgtacca ggagctgtac gactatgtga atcacagact gtcccacgac
     4681 ctgtctgatg aggccagggc cctgctgccc aacgtgatca ccaaggaggt gtctcacgag
     4741 atcatcaagg ataggcgctt taccagcgac aagttctttt tccacgtgcc tatcacactg
     4801 aactatcagg ccgccaattc cccatctaag ttcaaccaga gggtgaatgc ctacctgaag
     4861 gagcaccccg agacacctat catcggcatc gatcggggcg agagaaacct gatctatatc
     4921 acagtgatcg actccaccgg caagatcctg gagcagcgga gcctgaacac catccagcag
     4981 tttgattacc agaagaagct ggacaacagg gagaaggaga gggtggcagc aaggcaggcc
     5041 tggtctgtgg tgggcacaat caaggatctg aagcagggct atctgagcca ggtcatccac
     5101 gagatcgtgg acctgatgat ccactaccag gccgtggtgg tgctggagaa cctgaatttc
     5161 ggctttaaga gcaagaggac cggcatcgcc gagaaggccg tgtaccagca gttcgagaag
     5221 atgctgatcg ataagctgaa ttgcctggtg ctgaaggact atccagcaga gaaagtggga
     5281 ggcgtgctga acccatacca gctgacagac cagttcacct cctttgccaa gatgggcacc
     5341 cagtctggct tcctgtttta cgtgcctgcc ccatatacat ctaagatcga tcccctgacc
     5401 ggcttcgtgg accccttcgt gtggaaaacc atcaagaatc acgagagccg caagcacttc
     5461 ctggagggct tcgactttct gcactacgac gtgaaaaccg gcgacttcat cctgcacttt
     5521 aagatgaaca gaaatctgtc cttccagagg ggcctgcccg gctttatgcc tgcatgggat
     5581 atcgtgttcg agaagaacga gacacagttt gacgccaagg gcaccccttt catcgccggc
     5641 aagagaatcg tgccagtgat cgagaatcac agattcaccg gcagataccg ggacctgtat
     5701 cctgccaacg agctgatcgc cctgctggag gagaagggca tcgtgttcag ggatggctcc
     5761 aacatcctgc caaagctgct ggagaatgac gattctcacg ccatcgacac catggtggcc
     5821 ctgatccgca gcgtgctgca gatgcggaac tccaatgccg ccacaggcga ggactatatc
     5881 aacagccccg tgcgcgatct gaatggcgtg tgcttcgact cccggtttca gaacccagag
     5941 tggcccatgg acgccgatgc caatggcgcc taccacatcg ccctgaaggg ccagctgctg
     6001 ctgaatcacc tgaaggagag caaggatctg aagctgcaga acggcatctc caatcaggac
     6061 tggctggcct acatccagga gctgcgcaac ggcggctccc ctgctgctaa acgtgttaag
     6121 cttgatgggg gtagcccggc agccaagaga gtcaaactcg acggcggctc cccagcggca
     6181 aaaagggtga aactagacgg gggtagcccc gccgcgaagc gtgtaaagct ggatggagga
     6241 tcgcctgcgg ctaagcgagt caaattagac ggtgggtcac cggctgcaaa gcgcgtcaaa
     6301 ttggatgcta gcggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg
     6361 gaggagaacc ccggccccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     6421 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     6481 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     6541 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     6601 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     6661 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     6721 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc gcccgaccac
     6781 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     6841 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     6901 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     6961 cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     7021 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     7081 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     7141 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     7201 gtgtgcactg tgtttgctga cgcaaccccc actggttggg acattgccac cacctgtcag
     7261 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     7321 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     7381 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     7441 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     7501 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     7561 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     7621 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     7681 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     7741 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     7801 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     7861 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
     7921 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     7981 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     8041 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     8101 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     8161 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     8221 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     8281 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     8341 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     8401 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     8461 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     8521 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     8581 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     8641 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     8701 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     8761 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     8821 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     8881 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     8941 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     9001 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     9061 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     9121 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     9181 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     9241 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     9301 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     9361 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     9421 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     9481 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     9541 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     9601 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     9661 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     9721 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     9781 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     9841 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     9901 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     9961 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
    10021 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
    10081 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
    10141 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
    10201 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
    10261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
    10321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
    10381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
    10441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
    10501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
    10561 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
    10621 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
    10681 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
    10741 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
    10801 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
    10861 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
    10921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
    10981 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
    11041 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
    11101 accctcacta aagggaacaa aagctggagc tgcaagc
//
