LOCUS       sequence-486059-       14460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of dEnAsCas12a-4xSID with EGFP selectable 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14460)
  AUTHORS   Melore SM, Hamilton MC, Reddy TE
  TITLE     HyperCas12a enables highly-multiplexed epigenome editing screens.
  JOURNAL   bioRxiv [Preprint]. 2024 Jul 9:2024.07.08.602263. doi: 
            10.1101/2024.07.08.602263.
  PUBMED    39026853
REFERENCE   2  (bases 1 to 14460)
  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: "bioRxiv 
            [Preprint]. 2024 Jul 9:2024.07.08.602263. doi: 
            10.1101/2024.07.08.602263."
COMMENT     Sequence Label: SM279_EF1A-dEnAsCas12a-4xSID-T2A-EGFP
FEATURES             Location/Qualifiers
     source          1..14460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2630..3808
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2861..3799
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3756..3776
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3824..7744
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     CDS             7751..7777
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7757..7783
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             7796..7822
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7829..7855
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             8288..8314
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8318..8338
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8345..8398
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(8405..8407,8408..9118)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8405..8407,8408..9118)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8447..8468)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     9055..9076
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    9152..9740
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9205..9225)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9623..9634)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9743..9759)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9744..9760)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10265..10445
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10471..10488)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10477..10701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10747..11175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10834..10853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11044..11065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11184..11204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11189..11518
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11369..11504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11431..11450
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11566..11613
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11632..12006
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12136..12257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12173..12192)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12227..12246
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12306..12322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12306..12322)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12319..12341)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12330..12346
                     /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)."
     promoter        complement(join(12354..12360,12361..12378,12379..12384))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12399..12420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12537..12554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12708..13296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12788..12807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13467..14258,14259..14327))
                     /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     14090..14109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14328..14432)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta acccgtgtcg gctccagatc tggcctccgg gctccggtgc
     2641 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     2701 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     2761 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     2821 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     2881 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
     2941 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     3001 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     3061 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     3121 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     3181 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     3241 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     3301 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     3361 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     3421 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     3481 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     3541 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     3601 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     3661 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     3721 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     3781 tttttttctt ccatttcagg tgtcgtgagc ttctagagcc accatgacac agttcgaggg
     3841 ctttaccaac ctgtatcagg tgagcaagac actgcggttt gagctgatcc cacagggcaa
     3901 gaccctgaag cacatccagg agcagggctt catcgaggag gacaaggccc gcaatgatca
     3961 ctacaaggag ctgaagccca tcatcgatcg gatctacaag acctatgccg accagtgcct
     4021 gcagctggtg cagctggatt gggagaacct gagcgccgcc atcgactcct atagaaagga
     4081 gaaaaccgag gagacaagga acgccctgat cgaggagcag gccacatatc gcaatgccat
     4141 ccacgactac ttcatcggcc ggacagacaa cctgaccgat gccatcaata agagacacgc
     4201 cgagatctac aagggcctgt tcaaggccga gctgtttaat ggcaaggtgc tgaagcagct
     4261 gggcaccgtg accacaaccg agcacgagaa cgccctgctg cggagcttcg acaagtttac
     4321 aacctacttc tccggctttt atagaaacag gaagaacgtg ttcagcgccg aggatatcag
     4381 cacagccatc ccacaccgca tcgtgcagga caacttcccc aagtttaagg agaattgtca
     4441 catcttcaca cgcctgatca ccgccgtgcc cagcctgcgg gagcactttg agaacgtgaa
     4501 gaaggccatc ggcatcttcg tgagcacctc catcgaggag gtgttttcct tcccttttta
     4561 taaccagctg ctgacacaga cccagatcga cctgtataac cagctgctgg gaggaatctc
     4621 tcgggaggca ggcaccgaga agatcaaggg cctgaacgag gtgctgaatc tggccatcca
     4681 gaagaatgat gagacagccc acatcatcgc ctccctgcca cacagattca tccccctgtt
     4741 taagcagatc ctgtccgata ggaacaccct gtctttcatc ctggaggagt ttaagagcga
     4801 cgaggaagtg atccagtcct tctgcaagta caagacactg ctgagaaacg agaacgtgct
     4861 ggagacagcc gaggccctgt ttaacgagct gaacagcatc gacctgacac acatcttcat
     4921 cagccacaag aagctggaga caatcagcag cgccctgtgc gaccactggg atacactgag
     4981 gaatgccctg tatgagcgga gaatctccga gctgacaggc aagatcacca agtctgccaa
     5041 ggagaaggtg cagcgcagcc tgaagcacga ggatatcaac ctgcaggaga tcatctctgc
     5101 cgcaggcaag gagctgagcg aggccttcaa gcagaaaacc agcgagatcc tgtcccacgc
     5161 acacgccgcc ctggatcagc cactgcctac aaccctgaag aagcaggagg agaaggagat
     5221 cctgaagtct cagctggaca gcctgctggg cctgtaccac ctgctggact ggtttgccgt
     5281 ggatgagtcc aacgaggtgg accccgagtt ctctgcccgg ctgaccggca tcaagctgga
     5341 gatggagcct tctctgagct tctacaacaa ggccagaaat tatgccacca agaagcccta
     5401 ctccgtggag aagttcaagc tgaactttca gatgcctaca ctggccagag gctgggacgt
     5461 gaatagagag aagaacaatg gcgccatcct gtttgtgaag aacggcctgt actatctggg
     5521 catcatgcca aagcagaagg gcaggtataa ggccctgagc ttcgagccca cagagaaaac
     5581 cagcgagggc tttgataaga tgtactatga ctacttccct gatgccgcca agatgatccc
     5641 aaagtgcagc acccagctga aggccgtgac agcccacttt cagacccaca caacccccat
     5701 cctgctgtcc aacaatttca tcgagcctct ggagatcaca aaggagatct acgacctgaa
     5761 caatcctgag aaggagccaa agaagtttca gacagcctac gccaagaaaa ccggcgacca
     5821 gaagggctac agagaggccc tgtgcaagtg gatcgacttc acaagggatt ttctgtccaa
     5881 gtataccaag acaacctcta tcgatctgtc tagcctgcgg ccatcctctc agtataagga
     5941 cctgggcgag tactatgccg agctgaatcc cctgctgtac cacatcagct tccagagaat
     6001 cgccgagaag gagatcatgg atgccgtgga gacaggcaag ctgtacctgt tccagatcta
     6061 taacaaggac tttgccaagg gccaccacgg caagcctaat ctgcacacac tgtattggac
     6121 cggcctgttt tctccagaga acctggccaa gacaagcatc aagctgaatg gccaggccga
     6181 gctgttctac cgccctaagt ccaggatgaa gaggatggca caccggctgg gagagaagat
     6241 gctgaacaag aagctgaagg atcagaaaac cccaatcccc gacaccctgt accaggagct
     6301 gtacgactat gtgaatcaca gactgtccca cgacctgtct gatgaggcca gggccctgct
     6361 gcccaacgtg atcaccaagg aggtgtctca cgagatcatc aaggataggc gctttaccag
     6421 cgacaagttc tttttccacg tgcctatcac actgaactat caggccgcca attccccatc
     6481 taagttcaac cagagggtga atgcctacct gaaggagcac cccgagacac ctatcatcgg
     6541 catcgcccgg ggcgagagaa acctgatcta tatcacagtg atcgactcca ccggcaagat
     6601 cctggagcag cggagcctga acaccatcca gcagtttgat taccagaaga agctggacaa
     6661 cagggagaag gagagggtgg cagcaaggca ggcctggtct gtggtgggca caatcaagga
     6721 tctgaagcag ggctatctga gccaggtcat ccacgagatc gtggacctga tgatccacta
     6781 ccaggccgtg gtggtgctgg agaacctgaa tttcggcttt aagagcaaga ggaccggcat
     6841 cgccgagaag gccgtgtacc agcagttcga gaagatgctg atcgataagc tgaattgcct
     6901 ggtgctgaag gactatccag cagagaaagt gggaggcgtg ctgaacccat accagctgac
     6961 agaccagttc acctcctttg ccaagatggg cacccagtct ggcttcctgt tttacgtgcc
     7021 tgccccatat acatctaaga tcgatcccct gaccggcttc gtggacccct tcgtgtggaa
     7081 aaccatcaag aatcacgaga gccgcaagca cttcctggag ggcttcgact ttctgcacta
     7141 cgacgtgaaa accggcgact tcatcctgca ctttaagatg aacagaaatc tgtccttcca
     7201 gaggggcctg cccggcttta tgcctgcatg ggatatcgtg ttcgagaaga acgagacaca
     7261 gtttgacgcc aagggcaccc ctttcatcgc cggcaagaga atcgtgccag tgatcgagaa
     7321 tcacagattc accggcagat accgggacct gtatcctgcc aacgagctga tcgccctgct
     7381 ggaggagaag ggcatcgtgt tcagggatgg ctccaacatc ctgccaaagc tgctggagaa
     7441 tgacgattct cacgccatcg acacgatggt ggccctgatc cgcagcgtgc tgcagatgcg
     7501 gaactccaat gccgccacag gcgaggacta tatcaacagc cccgtgcgcg atctgaatgg
     7561 cgtgtgcttc gactcccggt ttcagaaccc agagtggcca atggacgccg atgccaatgg
     7621 cgcctaccac atcgccctga agggccagct gctgctgaat cacctgaagg agagcaagga
     7681 tctgaagctg cagaacggca tctccaatca ggactggctg gcctacatcc aggagctgcg
     7741 caacggatcc tatccctatg acgtgcccga ttatgccagc ctgggcagcg gctcccctgc
     7801 tgccaaacgc gttaaactag acgaagatcc tgccgcgaag cgcgtgaaac tggacgacgg
     7861 cattggtagt gggagcaacg gcagcagcct cggctccgga atgaacattc agatgttgct
     7921 cgaggctgct gattacctgg agcgccgcga acgggaagcg gaacacggtt acgctagtat
     7981 gctgcctggg agcggcatga atatccagat gctgctcgag gcggccgatt atctcgaacg
     8041 cagagagaga gaagccgaac atggctacgc ctctatgctg cctggctccg ggatgaatat
     8101 acagatgctc ctggaagctg ccgactatct ggagcgaaga gaaagggagg ctgagcacgg
     8161 gtatgcaagt atgctccctg gctccggtat gaatatccag atgctgctgg aggccgctga
     8221 ctatcttgaa cgccgagaaa gggaagccga gcatggatat gcatccatgc tgccttctag
     8281 gtctgcctac ccatacgatg ttccagatta cgcttcgccg aagaaaaagc gcaaggtcgc
     8341 tagcgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctgg
     8401 taccgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     8461 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     8521 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     8581 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     8641 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     8701 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     8761 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     8821 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     8881 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     8941 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     9001 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     9061 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagac
     9121 cggttgagaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     9181 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     9241 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     9301 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     9361 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     9421 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     9481 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     9541 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     9601 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     9661 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     9721 cctttgggcc gcctccccgc atcgataccg tcgacctcga gacctagaaa aacatggagc
     9781 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     9841 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     9901 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     9961 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
    10021 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
    10081 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
    10141 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
    10201 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
    10261 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
    10321 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
    10381 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
    10441 tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
    10501 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
    10561 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
    10621 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
    10681 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
    10741 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
    10801 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
    10861 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
    10921 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
    10981 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
    11041 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
    11101 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
    11161 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
    11221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
    11281 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
    11341 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
    11401 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
    11461 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
    11521 gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca
    11581 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
    11641 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
    11701 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
    11761 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
    11821 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
    11881 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
    11941 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
    12001 gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
    12061 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
    12121 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
    12181 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
    12241 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
    12301 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
    12361 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
    12421 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
    12481 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
    12541 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
    12601 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
    12661 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    12721 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    12781 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    12841 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    12901 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    12961 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    13021 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    13081 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    13141 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    13201 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    13261 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    13321 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
    13381 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
    13441 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
    13501 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
    13561 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
    13621 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    13681 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    13741 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    13801 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    13861 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    13921 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    13981 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    14041 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    14101 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    14161 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    14221 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    14281 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    14341 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    14401 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
//
