LOCUS       sequence-486072-       14223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of dHyperLbCas12a-4xSID with EGFP selectable 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14223)
  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 14223)
  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: SM270_EF1A-dHyperLbCas12a-4xSID-T2A-EGFP
FEATURES             Location/Qualifiers
     source          1..14223
                     /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             8051..8077
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8081..8101
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8108..8161
                     /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(8168..8170,8171..8881)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8168..8170,8171..8881)
                     /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(8210..8231)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     8818..8839
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8915..9503
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8968..8988)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9386..9397)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9506..9522)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9507..9523)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10028..10208
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10234..10251)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10240..10464
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10510..10938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10597..10616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10807..10828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10947..10967)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10952..11281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11132..11267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11194..11213
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11329..11376
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11395..11769
                     /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    11899..12020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11936..11955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11990..12009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12069..12085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12069..12085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12082..12104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12093..12109
                     /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(12117..12123,12124..12141,12142..12147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12162..12183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12300..12317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12471..13059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12551..12570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13230..14021,14022..14090))
                     /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     13853..13872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14091..14195)
                     /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 cctcggctcc ggaatgaaca ttcagatgtt
     7681 gctcgaggct gctgattacc tggagcgccg cgaacgggaa gcggaacacg gttacgctag
     7741 tatgctgcct gggagcggca tgaatatcca gatgctgctc gaggcggccg attatctcga
     7801 acgcagagag agagaagccg aacatggcta cgcctctatg ctgcctggct ccgggatgaa
     7861 tatacagatg ctcctggaag ctgccgacta tctggagcga agagaaaggg aggctgagca
     7921 cgggtatgca agtatgctcc ctggctccgg tatgaatatc cagatgctgc tggaggccgc
     7981 tgactatctt gaacgccgag aaagggaagc cgagcatgga tatgcatcca tgctgccttc
     8041 taggtctgcc tacccatacg atgttccaga ttacgcttcg ccgaagaaaa agcgcaaggt
     8101 cgctagcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc
     8161 tggtaccgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     8221 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     8281 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     8341 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     8401 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     8461 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     8521 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     8581 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     8641 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     8701 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     8761 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     8821 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     8881 gaccggttga gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     8941 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     9001 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     9061 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     9121 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     9181 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     9241 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     9301 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     9361 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     9421 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     9481 ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagacctag aaaaacatgg
     9541 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
     9601 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
     9661 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     9721 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt
     9781 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg
     9841 gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa
     9901 cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt
     9961 agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga
    10021 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
    10081 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
    10141 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
    10201 ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
    10261 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
    10321 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
    10381 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
    10441 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
    10501 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
    10561 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
    10621 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
    10681 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
    10741 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
    10801 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
    10861 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
    10921 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
    10981 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
    11041 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
    11101 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
    11161 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
    11221 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
    11281 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
    11341 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
    11401 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
    11461 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
    11521 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
    11581 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
    11641 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
    11701 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
    11761 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
    11821 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
    11881 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    11941 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    12001 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
    12061 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    12121 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    12181 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    12241 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    12301 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    12361 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    12421 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    12481 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    12541 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    12601 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    12661 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    12721 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    12781 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    12841 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    12901 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    12961 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    13021 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    13081 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    13141 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    13201 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    13261 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    13321 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    13381 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    13441 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    13501 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    13561 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    13621 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    13681 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    13741 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    13801 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    13861 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    13921 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    13981 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    14041 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    14101 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    14161 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    14221 gac
//
