LOCUS       sequence-486083-       15345 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Mammalian expression of dHyperLbCas12a-VPR with EGFP selectable 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15345)
  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 15345)
  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: SM272_EF1A-dHyperLbCas12a-VPR-T2A-EGFP
FEATURES             Location/Qualifiers
     source          1..15345
                     /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..7507
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFRNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIRVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGRGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKRIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             7514..7540
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7520..7546
                     /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             7559..7585
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7592..7618
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7655..7804
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             7829..7849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8279..8635
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             8654..9223
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             9230..9283
                     /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(9290..9292,9293..10003)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(9290..9292,9293..10003)
                     /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(9332..9353)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     9940..9961
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    10037..10625
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10090..10110)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10508..10519)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10628..10644)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10629..10645)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11150..11330
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11356..11373)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11362..11586
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11632..12060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11719..11738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11929..11950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12069..12089)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12074..12403
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12254..12389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12316..12335
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12451..12498
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12517..12891
                     /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    13021..13142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13058..13077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13112..13131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13191..13207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13191..13207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13204..13226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13215..13231
                     /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(13239..13245,13246..13263,13264..13269))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13284..13305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13422..13439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13593..14181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13673..13692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14352..15143,15144..15212))
                     /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     14975..14994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15213..15317)
                     /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 accatgagca agctggagaa
     3841 gtttacaaac tgctactccc tgtctaagac cctgaggttc aaggccatcc ctgtgggcaa
     3901 gacccaggag aacatcgaca ataagcggct gctggtggag gacgagaaga gagccgagga
     3961 ttataagggc gtgaagaagc tgctggatcg ctactatctg tcttttatca acgacgtgct
     4021 gcacagcatc aagctgaaga atctgaacaa ttacatcagc ctgttccgga agaaaaccag
     4081 aaccgagaag gagaataagg agctggagaa cctggagatc aatctgcgga aggagatcgc
     4141 caaggccttc aagggcaacg agggctacaa gtccctgttt aagaaggata tcatcgagac
     4201 aatcctgcca gagttcctgg acgataagga cgagatcgcc ctggtgaaca gcttcaatgg
     4261 ctttaccaca gccttcaccg gcttctttcg caacagagag aatatgtttt ccgaggaggc
     4321 caagagcaca tccatcgcct tcaggtgtat caacgagaat ctgacccgct acatctctaa
     4381 tatggacatc ttcgagaagg tggacgccat ctttgataag cacgaggtgc aggagatcaa
     4441 ggagaagatc ctgaacagcg actatgatgt ggaggatttc tttgagggcg agttctttaa
     4501 ctttgtgctg acacaggagg gcatccgcgt gtataacgcc atcatcggcg gcttcgtgac
     4561 cgagagcggc gagaagatca agggcctgaa cgagtacatc aacctgtata atcagaaaac
     4621 caagcagaag ctgcctaagt ttaagccact gtataagcag gtgctgagcg atcgggagtc
     4681 tctgagcttc tacggccggg gctatacatc cgatgaggag gtgctggagg tgtttagaaa
     4741 caccctgaac aagaacagcg agatcttcag ctccatcaag aagctggaga agctgttcaa
     4801 gaattttgac gagtactcta gcgccggcat ctttgtgaag aacggccccg ccatcagcac
     4861 aatctccaag cgcatcttcg gcgagtggaa cgtgatccgg gacaagtgga atgccgagta
     4921 tgacgatatc cacctgaaga agaaggccgt ggtgaccgag aagtacgagg acgatcggag
     4981 aaagtccttc aagaagatcg gctccttttc tctggagcag ctgcaggagt acgccgacgc
     5041 cgatctgtct gtggtggaga agctgaagga gatcatcatc cagaaggtgg atgagatcta
     5101 caaggtgtat ggctcctctg agaagctgtt cgacgccgat tttgtgctgg agaagagcct
     5161 gaagaagaac gacgccgtgg tggccatcat gaaggacctg ctggattctg tgaagagctt
     5221 cgagaattac atcaaggcct tctttggcga gggcaaggag acaaacaggg acgagtcctt
     5281 ctatggcgat tttgtgctgg cctacgacat cctgctgaag gtggaccaca tctacgatgc
     5341 catccgcaat tatgtgaccc agaagcccta ctctaaggat aagttcaagc tgtattttca
     5401 gaaccctcag ttcatgggcg gctgggacaa ggataaggag acagactatc gggccaccat
     5461 cctgagatac ggctccaagt actatctggc catcatggat aagaagtacg ccaagtgcct
     5521 gcagaagatc gacaaggacg atgtgaacgg caattacgag aagatcaact ataagctgct
     5581 gcccggccct aataagatgc tgccaaaggt gttcttttct aagaagtgga tggcctacta
     5641 taaccccagc gaggacatcc agaagatcta caagaatggc acattcaaga agggcgatat
     5701 gtttaacctg aatgactgtc acaagctgat cgacttcttt aaggatagca tctcccggta
     5761 tccaaagtgg tccaatgcct acgatttcaa cttttctgag acagagaagt ataaggacat
     5821 cgccggcttt tacagagagg tggaggagca gggctataag gtgagcttcg agtctgccag
     5881 caagaaggag gtggataagc tggtggagga gggcaagctg tatatgttcc agatctataa
     5941 caaggacttt tccgataagt ctcacggcac acccaatctg cacaccatgt acttcaagct
     6001 gctgtttgac gagaacaatc acggacagat caggctgagc ggaggagcag agctgttcat
     6061 gaggcgcgcc tccctgaaga aggaggagct ggtggtgcac ccagccaact cccctatcgc
     6121 caacaagaat ccagataatc ccaagaaaac cacaaccctg tcctacgacg tgtataagga
     6181 taagaggttt tctgaggacc agtacgagct gcacatccca atcgccatca ataagtgccc
     6241 caagaacatc ttcaagatca atacagaggt gcgcgtgctg ctgaagcacg acgataaccc
     6301 ctatgtgatc ggcatcgcca ggggcgagcg caatctgctg tatatcgtgg tggtggacgg
     6361 caagggcaac atcgtggagc agtattccct gaacgagatc atcaacaact tcaacggcat
     6421 caggatcaag acagattacc actctctgct ggacaagaag gagaaggaga ggttcgaggc
     6481 ccgccagaac tggacctcca tcgagaatat caaggagctg aaggccggct atatctctca
     6541 ggtggtgcac aagatctgcg agctggtgga gaagtacgat gccgtgatcg ccctggagga
     6601 cctgaactct ggctttaaga atagccgcgt gaaggtggag aagcaggtgt atcagaagtt
     6661 cgagaagatg ctgatcgata agctgaacta catggtggac aagaagtcta atccttgtgc
     6721 aacaggcggc gccctgaagg gctatcagat caccaataag ttcgagagct ttaagtccat
     6781 gtctacccag aacggcttca tcttttacat ccctgcctgg ctgacatcca agatcgatcc
     6841 atctaccggc tttgtgaacc tgctgaaaac caagtatacc agcatcgccg attccaagaa
     6901 gttcatcagc tcctttgaca ggatcatgta cgtgcccgag gaggatctgt tcgagtttgc
     6961 cctggactat aagaacttct ctcgcacaga cgccgattac atcaagaagt ggaagctgta
     7021 ctcctacggc aaccggatca gaatcttccg gaatcctaag aagaacaacg tgttcgactg
     7081 ggaggaggtg tgcctgacca gcgcctataa ggagctgttc aacaagtacg gcatcaatta
     7141 tcagcagggc gatatcagag ccctgctgtg cgagcagtcc gacaaggcct tctactctag
     7201 ctttatggcc ctgatgagcc tgatgctgca gatgcggaac agcatcacag gccgcaccga
     7261 cgtggatttt ctgatcagcc ctgtgaagaa ctccgacggc atcttctacg atagccggaa
     7321 ctatgaggcc caggagaatg ccatcctgcc aaagaacgcc gacgccaatg gcgcctataa
     7381 catcgccaga aaggtgctgt gggccatcgg ccagttcaag aaggccgagg acgagaagct
     7441 ggataaggtg aagatcgcca tctctaacaa ggagtggctg gagtacgccc agaccagcgt
     7501 gaagcacgga tcctatccct atgacgtgcc cgattatgcc agcctgggca gcggctcccc
     7561 tgctgccaaa cgcgttaaac tagacgaaga tcctgccgcg aagcgcgtga aactggacga
     7621 cggcattggt agtgggagca acggcagcag cctcgacgca ttggacgatt ttgatctgga
     7681 tatgctggga agtgacgccc tcgatgattt tgaccttgac atgcttggtt cggatgccct
     7741 tgatgacttt gacctcgaca tgctcggcag tgacgccctt gatgatttcg acctggacat
     7801 gctgattaac tctagaagtt ccggatctcc gaaaaagaaa cgcaaagttg gtagccagta
     7861 cctgcccgac accgacgacc ggcaccggat cgaggaaaag cggaagcgga cctacgagac
     7921 attcaagagc atcatgaaga agtccccctt cagcggcccc accgacccta gacctccacc
     7981 tagaagaatc gccgtgccca gcagatccag cgccagcgtg ccaaaacctg ccccccagcc
     8041 ttaccccttc accagcagcc tgagcaccat caactacgac gagttcccta ccatggtgtt
     8101 ccccagcggc cagatctctc aggcctctgc tctggctcca gcccctcctc aggtgctgcc
     8161 tcaggctcct gctcctgcac cagctccagc catggtgtct gcactggctc aggcaccagc
     8221 acccgtgcct gtgctggctc ctggacctcc acaggctgtg gctccaccag cccctaaacc
     8281 tacacaggcc ggcgagggca cactgtctga agctctgctg cagctgcagt tcgacgacga
     8341 ggatctggga gccctgctgg gaaacagcac cgatcctgcc gtgttcaccg acctggccag
     8401 cgtggacaac agcgagttcc agcagctgct gaaccagggc atccctgtgg cccctcacac
     8461 caccgagccc atgctgatgg aataccccga ggccatcacc cggctcgtga caggcgctca
     8521 gaggcctcct gatccagctc ctgcccctct gggagcacca ggcctgccta atggactgct
     8581 gtctggcgac gaggacttca gctctatcgc cgatatggat ttctcagcct tgctgggctc
     8641 tggcagcggc agccgggatt ccagggaagg gatgtttttg ccgaagcctg aggccggctc
     8701 cgctattagt gacgtgtttg agggccgcga ggtgtgccag ccaaaacgaa tccggccatt
     8761 tcatcctcca ggaagtccat gggccaaccg cccactcccc gccagcctcg caccaacacc
     8821 aaccggtcca gtacatgagc cagtcgggtc actgaccccg gcaccagtcc ctcagccact
     8881 ggatccagcg cccgcagtga ctcccgaggc cagtcacctg ttggaggatc ccgatgaaga
     8941 gacgagccag gctgtcaaag cccttcggga gatggccgat actgtgattc cccagaagga
     9001 agaggctgca atctgtggcc aaatggacct ttcccatccg cccccaaggg gccatctgga
     9061 tgagctgaca accacacttg agtccatgac cgaggatctg aacctggact cacccctgac
     9121 cccggaattg aacgagattc tggatacctt cctgaacgac gagtgcctct tgcatgccat
     9181 gcatatcagc acaggactgt ccatcttcga cacatctctg tttgctagcg agggcagagg
     9241 aagtcttcta acatgcggtg acgtggagga gaatcccggc cctggtaccg tgagcaaggg
     9301 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     9361 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     9421 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     9481 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     9541 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     9601 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     9661 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     9721 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     9781 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     9841 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     9901 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     9961 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagaccggtt gagaattcga
    10021 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
    10081 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
    10141 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
    10201 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
    10261 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
    10321 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
    10381 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
    10441 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
    10501 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
    10561 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
    10621 cccgcatcga taccgtcgac ctcgagacct agaaaaacat ggagcaatca caagtagcaa
    10681 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
    10741 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
    10801 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
    10861 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
    10921 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
    10981 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
    11041 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
    11101 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
    11161 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    11221 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    11281 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
    11341 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
    11401 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
    11461 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
    11521 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
    11581 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
    11641 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
    11701 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    11761 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
    11821 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
    11881 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    11941 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
    12001 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
    12061 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
    12121 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
    12181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
    12241 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
    12301 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
    12361 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
    12421 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
    12481 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
    12541 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
    12601 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
    12661 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
    12721 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
    12781 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
    12841 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
    12901 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
    12961 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
    13021 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    13081 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    13141 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
    13201 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
    13261 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
    13321 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
    13381 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
    13441 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
    13501 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    13561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    13621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    13681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    13741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    13801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    13861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    13921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    13981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    14041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    14101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    14161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    14221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    14281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    14341 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    14401 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    14461 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    14521 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    14581 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    14641 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    14701 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    14761 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    14821 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    14881 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    14941 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    15001 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    15061 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    15121 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    15181 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    15241 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    15301 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac
//
