LOCUS       sequence-326356-       14988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one lentiviral CRISPRa gRNA expression with blasticidin 
            selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14988)
  AUTHORS   Chan W, Gottschalk RA, Yao Y, Pomerantz JL, Germain RN
  TITLE     Efficient Immune Cell Genome Engineering with Enhanced CRISPR 
            Editing Tools.
  JOURNAL   Immunohorizons. 2021 Feb 23;5(2):117-132. doi: 
            10.4049/immunohorizons.2000082.
  PUBMED    33622708
REFERENCE   2  (bases 1 to 14988)
  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"; doi: 
            "10.4049/immunohorizons"; journalName: "Immunohorizons"; date: 
            "2021-02-23- 23"; volume: "5"; issue: "2"; pages: "117-132"
COMMENT     Sequence Label: LentiSAMPHv2
FEATURES             Location/Qualifiers
     source          1..14988
                     /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        2607..2855
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2607..2627
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2778..2797
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2893..2911
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        2963..2981
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        3068..3279
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3296..3343
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3344..3367,3368..3370,3371..5926,5927..5929,
                     5930..7444)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7499..7519
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7544..7693
                     /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             7715..7771
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7772..8158
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             8213..8233
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8249..8791
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             8816..9187
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             9203..9256
                     /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             9257..9655
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9680..10268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9733..9753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10151..10162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10271..10287)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10272..10288)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10793..10973
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10999..11016)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11005..11229
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11275..11703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11362..11381)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11572..11593
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11712..11732)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11717..12046
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11897..12032
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11959..11978
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12094..12141
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12160..12534
                     /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    12664..12785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12701..12720)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12755..12774
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12834..12850)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12834..12850)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12847..12869)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12858..12874
                     /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(12882..12888,12889..12906,12907..12912))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12927..12948
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13065..13082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13236..13824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13316..13335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13995..14786,14787..14855))
                     /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     14618..14637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14856..14960)
                     /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 gctagcgagg gcctatttcc catgattcct tcatatttgc
     2641 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     2701 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     2761 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     2821 tggctttata tatcttgtgg aaaggacgaa acaccggaga cgggataccg tctctgtttt
     2881 agagctaggc caacatgagg atcacccatg tctgcagggc ctagcaagtt aaaataaggc
     2941 tagtccgtta tcaacttggc caacatgagg atcacccatg tctgcagggc caagtggcac
     3001 cgagtcggtg ctttttttgg atcctaggtc ttgaaaggag tgggaattgg ctccggtgcc
     3061 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     3121 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     3181 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     3241 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg cgtacggcca ccatgaaaag
     3301 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggacaaga agtacagcat
     3361 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     3421 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     3481 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     3541 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     3601 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     3661 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     3721 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     3781 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     3841 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     3901 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     3961 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     4021 ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg
     4081 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     4141 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     4201 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     4261 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     4321 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     4381 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     4441 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     4501 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     4561 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     4621 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     4681 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     4741 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     4801 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     4861 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     4921 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     4981 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     5041 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     5101 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     5161 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     5221 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     5281 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     5341 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     5401 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     5461 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     5521 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     5581 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     5641 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     5701 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     5761 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     5821 actggacatc aaccggctgt ccgactacga tgtggaccac atcgtgcctc agagctttct
     5881 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaaggccc ggggcaagag
     5941 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     6001 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     6061 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     6121 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     6181 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     6241 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     6301 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     6361 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     6421 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     6481 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     6541 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     6601 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     6661 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     6721 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
     6781 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     6841 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     6901 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     6961 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
     7021 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     7081 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     7141 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     7201 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     7261 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     7321 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
     7381 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     7441 cgacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag gtagcggacc
     7501 taagaaaaag aggaaggtgg cggccgctgg atccggacgg gctgacgcat tggacgattt
     7561 tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc
     7621 ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga
     7681 cctggacatg ctgattaact gtacagggtc cggcgcaaca aacttctctc tgctgaaaca
     7741 agccggagat gtcgaagaga atcctggacc ggcttcaaac tttactcagt tcgtgctcgt
     7801 ggacaatggt gggacagggg atgtgacagt ggctccttct aatttcgcta atggggtggc
     7861 agagtggatc agctccaact cacggagcca ggcctacaag gtgacatgca gcgtcaggca
     7921 gtctagtgcc cagaagagaa agtataccat caaggtggag gtccccaaag tggctaccca
     7981 gacagtgggc ggagtcgaac tgcctgtcgc cgcttggagg tcctacctga acatggagct
     8041 cactatccca attttcgcta ccaattctga ctgtgaactc atcgtgaagg caatgcaggg
     8101 gctcctcaaa gacggtaatc ctatcccttc cgccatcgcc gctaactcag gtatctacag
     8161 cgctggagga ggtggaagcg gaggaggagg aagcggagga ggaggtagcg gacctaagaa
     8221 aaagaggaag gtggcggccg ctggatcccc ttcagggcag atcagcaacc aggccctggc
     8281 tctggcccct agctccgctc cagtgctggc ccagactatg gtgccctcta gtgctatggt
     8341 gcctctggcc cagccacctg ctccagcccc tgtgctgacc ccaggaccac cccagtcact
     8401 gagcgctcca gtgcccaagt ctacacaggc cggcgagggg actctgagtg aagctctgct
     8461 gcacctgcag ttcgacgctg atgaggacct gggagctctg ctggggaaca gcaccgatcc
     8521 cggagtgttc acagatctgg cctccgtgga caactctgag tttcagcagc tgctgaatca
     8581 gggcgtgtcc atgtctcata gtacagccga accaatgctg atggagtacc ccgaagccat
     8641 tacccggctg gtgaccggca gccagcggcc ccccgacccc gctccaactc ccctgggaac
     8701 cagcggcctg cctaatgggc tgtccggaga tgaagacttc tcaagcatcg ctgatatgga
     8761 ctttagtgcc ctgctgtcac agatttcctc tagtgggcag ggaggaggtg gaagcggctt
     8821 cagcgtggac accagtgccc tgctggacct gttcagcccc tcggtgaccg tgcccgacat
     8881 gagcctgcct gaccttgaca gcagcctggc cagtatccaa gagctcctgt ctccccagga
     8941 gccccccagg cctcccgagg cagagaacag cagcccggat tcagggaagc agctggtgca
     9001 ctacacagcg cagccgctgt tcctgctgga ccccggctcc gtggacaccg ggagcaacga
     9061 cctgccggtg ctgtttgagc tgggagaggg ctcctacttc tccgaagggg acggcttcgc
     9121 cgaggacccc accatctccc tgctgacagg ctcggagcct cccaaagcca aggaccccac
     9181 tgtctcctgt acaggcagtg gagagggcag aggaagtctg ctaacatgcg gtgacgtcga
     9241 ggagaatcct ggcccaatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     9301 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     9361 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     9421 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     9481 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     9541 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     9601 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagaatt
     9661 cgatatcaag cttatcggta atcaacctct ggattacaaa atttgtgaaa gattgactgg
     9721 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     9781 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     9841 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     9901 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     9961 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
    10021 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
    10081 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
    10141 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
    10201 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
    10261 ctccccgcat cgataccgtc gacctcgaga cctagaaaaa catggagcaa tcacaagtag
    10321 caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt
    10381 gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga
    10441 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
    10501 acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa
    10561 ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt
    10621 accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca
    10681 ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga
    10741 cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct
    10801 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
    10861 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
    10921 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg
    10981 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
    11041 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
    11101 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
    11161 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
    11221 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
    11281 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
    11341 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
    11401 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
    11461 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
    11521 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
    11581 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
    11641 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
    11701 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
    11761 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
    11821 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
    11881 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
    11941 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
    12001 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
    12061 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
    12121 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
    12181 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
    12241 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
    12301 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
    12361 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
    12421 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
    12481 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
    12541 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
    12601 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
    12661 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
    12721 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    12781 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
    12841 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
    12901 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
    12961 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
    13021 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
    13081 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
    13141 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
    13201 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
    13261 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
    13321 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
    13381 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
    13441 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
    13501 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
    13561 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    13621 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
    13681 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    13741 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    13801 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
    13861 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
    13921 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
    13981 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
    14041 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
    14101 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
    14161 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
    14221 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
    14281 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
    14341 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
    14401 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
    14461 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
    14521 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
    14581 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
    14641 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
    14701 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
    14761 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
    14821 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
    14881 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
    14941 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
