LOCUS       sequence-486065-       15864 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Mammalian expression of dEnAsCas12a-P300 with EGFP selectable 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15864)
  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 15864)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 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: SM281_EF1A-dEnAsCas12a-P300-T2A-EGFP
FEATURES             Location/Qualifiers
     source          1..15864
                     /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             7892..9742
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      9506..9515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9749..9802
                     /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(9809..9811,9812..10522)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(9809..9811,9812..10522)
                     /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(9851..9872)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     10459..10480
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    10556..11144
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10609..10629)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11027..11038)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11147..11163)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11148..11164)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11669..11849
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11875..11892)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11881..12105
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12151..12579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12238..12257)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12448..12469
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12588..12608)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12593..12922
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12773..12908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12835..12854
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12970..13017
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             13036..13410
                     /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    13540..13661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13577..13596)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13631..13650
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13710..13726)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13710..13726)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13723..13745)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13734..13750
                     /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(13758..13764,13765..13782,13783..13788))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13803..13824
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13941..13958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14112..14700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14192..14211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14871..15662,15663..15731))
                     /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     15494..15513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15732..15836)
                     /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 cattttcaaa ccagaagaac tacgacaggc
     7921 actgatgcca actttggagg cactttaccg tcaggatcca gaatcccttc cctttcgtca
     7981 acctgtggac cctcagcttt taggaatccc tgattacttt gatattgtga agagccccat
     8041 ggatctttct accattaaga ggaagttaga cactggacag tatcaggagc cctggcagta
     8101 tgtcgatgat atttggctta tgttcaataa tgcctggtta tataaccgga aaacatcacg
     8161 ggtatacaaa tactgctcca agctctctga ggtctttgaa caagaaattg acccagtgat
     8221 gcaaagcctt ggatactgtt gtggcagaaa gttggagttc tctccacaga cactgtgttg
     8281 ctacggcaaa cagttgtgca caatacctcg tgatgccact tattacagtt accagaacag
     8341 gtatcatttc tgtgagaagt gtttcaatga gatccaaggg gagagcgttt ctttggggga
     8401 tgacccttcc cagcctcaaa ctacaataaa taaagaacaa ttttccaaga gaaaaaatga
     8461 cacactggat cctgaactgt ttgttgaatg tacagagtgc ggaagaaaga tgcatcagat
     8521 ctgtgtcctt caccatgaga tcatctggcc tgctggattc gtctgtgatg gctgtttaaa
     8581 gaaaagtgca cgaactagga aagaaaataa gttttctgct aaaaggttgc catctaccag
     8641 acttggcacc tttctagaga atcgtgtgaa tgactttctg aggcgacaga atcaccctga
     8701 gtcaggagag gtcactgtta gagtagttca tgcttctgac aaaaccgtgg aagtaaaacc
     8761 aggcatgaaa gcaaggtttg tggacagtgg agagatggca gaatcctttc cataccgaac
     8821 caaagccctc tttgcctttg aagaaattga tggtgttgac ctgtgcttct ttggcatgca
     8881 tgttcaagag tatggctctg actgccctcc acccaaccag aggagagtat acatatctta
     8941 cctcgatagt gttcatttct tccgtcctaa atgcttgagg actgcagtct atcatgaaat
     9001 cctaattgga tatttagaat atgtcaagaa attaggttac acaacagggc atatttgggc
     9061 atgtccacca agtgagggag atgattatat cttccattgc catcctcctg accagaagat
     9121 acccaagccc aagcgactgc aggaatggta caaaaaaatg cttgacaagg ctgtatcaga
     9181 gcgtattgtc catgactaca aggatatttt taaacaagct actgaagata gattaacaag
     9241 tgcaaaggaa ttgccttatt tcgagggtga tttctggccc aatgttctgg aagaaagcat
     9301 taaggaactg gaacaggagg aagaagagag aaaacgagag gaaaacacca gcaatgaaag
     9361 cacagatgtg accaagggag acagcaaaaa tgctaaaaag aagaataata agaaaaccag
     9421 caaaaataag agcagcctga gtaggggcaa caagaagaaa cccgggatgc ccaatgtatc
     9481 taacgacctc tcacagaaac tatatgccac catggagaag cataaagagg tcttctttgt
     9541 gatccgcctc attgctggcc ctgctgccaa ctccctgcct cccattgttg atcctgatcc
     9601 tctcatcccc tgcgatctga tggatggtcg ggatgcgttt ctcacgctgg caagggacaa
     9661 gcacctggag ttctcttcac tccgaagagc ccagtggtcc accatgtgca tgctggtgga
     9721 gctgcacacg cagagccagg acgctagcga gggcagagga agtcttctaa catgcggtga
     9781 cgtggaggag aatcccggcc ctggtaccgt gagcaagggc gaggagctgt tcaccggggt
     9841 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     9901 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     9961 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
    10021 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
    10081 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
    10141 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
    10201 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
    10261 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
    10321 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
    10381 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
    10441 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
    10501 cggcatggac gagctgtaca agaccggttg agaattcgat atcaagctta tcgataatca
    10561 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
    10621 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
    10681 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
    10741 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
    10801 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
    10861 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
    10921 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
    10981 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
    11041 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
    11101 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
    11161 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
    11221 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
    11281 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
    11341 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
    11401 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
    11461 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
    11521 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
    11581 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
    11641 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
    11701 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    11761 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    11821 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
    11881 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
    11941 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
    12001 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
    12061 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
    12121 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
    12181 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    12241 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    12301 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    12361 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    12421 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    12481 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    12541 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
    12601 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    12661 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    12721 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
    12781 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    12841 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
    12901 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
    12961 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    13021 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    13081 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    13141 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    13201 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    13261 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    13321 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    13381 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    13441 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    13501 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    13561 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    13621 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    13681 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    13741 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    13801 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
    13861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
    13921 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    13981 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    14041 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    14101 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    14161 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    14221 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    14281 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    14341 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    14401 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    14461 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    14521 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    14581 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    14641 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    14701 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    14761 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    14821 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    14881 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    14941 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    15001 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    15061 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    15121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    15181 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    15241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    15301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    15361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    15421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    15481 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    15541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    15601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    15661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    15721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    15781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    15841 tttccccgaa aagtgccacc tgac
//
