LOCUS       sequence-488051-       13981 bp    DNA     circular SYN 02-OCT-2025
DEFINITION  Cas12a CRISPRko all-in-one positive control; targets CD47, CD63.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13981)
  TITLE     Doench and Root CRISPR plasmids 2025
REFERENCE   2  (bases 1 to 13981)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pRDB_031
FEATURES             Location/Qualifiers
     source          1..13981
                     /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        complement(2761..3009)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2819..2838)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2989..3009)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        3083..4261
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3314..4252
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4209..4229
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4266..4275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4311..4331
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4338..8255
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      7975..7984
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8262..8309
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8316..8396
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             8403..8459
                     /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             8460..9053
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     8953..8973
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9073..9661
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9126..9146)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9544..9555)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9733..9966
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9992..10009)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9998..10222
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10268..10696
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10355..10374)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10565..10586
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10705..10725)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10710..11039
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10890..11025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10952..10971
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11087..11134
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11153..11527
                     /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    11657..11778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11694..11713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11748..11767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11827..11843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11827..11843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11840..11862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11851..11867
                     /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(11875..11881,11882..11899,11900..11905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11920..11941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12058..12075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12229..12817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12309..12328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12988..13779,13780..13848))
                     /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     13611..13630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13849..13953)
                     /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 aggtaccggg cagaaccgac tcggtgccac tttttcaaga
     2641 ccaggctata gctctcgtct atattcaaaa aagctgctaa ctacacagat tgggaatcta
     2701 cgatagtaga aattactgtt ctggggacag tttggtatat ctacaagagt agaaattacc
     2761 ggtgtttcgt cctttccaca agatatataa agccaagaaa tcgaaatact ttcaagttac
     2821 ggtaagcata tgatagtcca ttttaaaaca taattttaaa actgcaaact acccaagaaa
     2881 ttattacttt ctacgtcacg tattttgtac taatatcttt gtgtttacag tcaaattaat
     2941 tctaattatc tctctaacag ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa
     3001 taggccctct tcgaatctag ttgcaaagat ggataaagtt ttaaacagag aggaatcttt
     3061 gcagctaatg gaccttctac acggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     3121 cacagtcccc gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg
     3181 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     3241 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     3301 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
     3361 tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga
     3421 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
     3481 tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
     3541 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     3601 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
     3661 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     3721 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     3781 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     3841 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     3901 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     3961 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     4021 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     4081 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     4141 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     4201 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     4261 aaggagccac catggccaaa cggacagccg acggaagcga gttcgagtca ccaaagaaga
     4321 agcggaaagt cggcagcaca cagttcgagg gctttaccaa cctgtatcag gtgagcaaga
     4381 cactgcggtt tgagctgatc ccacagggca agaccctgaa gcacatccag gagcagggct
     4441 tcatcgagga ggacaaggcc cgcaatgatc actacaagga gctgaagccc atcatcgatc
     4501 ggatctacaa gacctatgcc gaccagtgcc tgcagctggt gcagctggat tgggagaacc
     4561 tgagcgccgc catcgactcc tatagaaagg agaaaaccga ggagacaagg aacgccctga
     4621 tcgaggagca ggccacatat cgcaatgcca tccacgacta cttcatcggc cggacagaca
     4681 acctgaccga tgccatcaat aagagacacg ccgagatcta caagggcctg ttcaaggccg
     4741 agctgtttaa tggcaaggtg ctgaagcagc tgggcaccgt gaccacaacc gagcacgaga
     4801 acgccctgct gcggagcttc gacaagttta caacctactt ctccggcttt tatcggaaca
     4861 ggaagaacgt gttcagcgcc gaggatatca gcacagccat cccacaccgc atcgtgcagg
     4921 acaacttccc caagtttaag gagaattgtc acatcttcac acgcctgatc accgccgtgc
     4981 ccagcctgcg ggagcacttt gagaacgtga agaaggccat cggcatcttc gtgagcacct
     5041 ccatcgagga ggtgttttcc ttcccttttt ataaccagct gctgacacag acccagatcg
     5101 acctgtataa ccagctgctg ggaggaatct ctcgggaggc aggcaccgag aagatcaagg
     5161 gcctgaacga ggtgctgaat ctggccatcc agaagaatga tgagacagcc cacatcatcg
     5221 cctccctgcc acacagattc atccccctgt ttaagcagat cctgtccgat aggaacaccc
     5281 tgtctttcat cctggaggag tttaagagcg acgaggaagt gatccagtcc ttctgcaagt
     5341 acaagacact gctgagaaac gagaacgtgc tggagacagc cgaggccctg tttaacgagc
     5401 tgaacagcat cgacctgaca cacatcttca tcagccacaa gaagctggag acaatcagca
     5461 gcgccctgtg cgaccactgg gatacactga ggaatgccct gtatgagcgg agaatctccg
     5521 agctgacagg caagatcacc aagtctgcca aggagaaggt gcagcgcagc ctgaagcacg
     5581 aggatatcaa cctgcaggag atcatctctg ccgcaggcaa ggagctgagc gaggccttca
     5641 agcagaaaac cagcgagatc ctgtcccacg cacacgccgc cctggatcag ccactgccta
     5701 caaccctgaa gaagcaggag gagaaggaga tcctgaagtc tcagctggac agcctgctgg
     5761 gcctgtacca cctgctggac tggtttgccg tggatgagtc caacgaggtg gaccccgagt
     5821 tctctgcccg gctgaccggc atcaagctgg agatggagcc ttctctgagc ttctacaaca
     5881 aggccagaaa ttatgccacc aagaagccct actccgtgga gaagttcaag ctgaactttc
     5941 agatgcctac actggcccgc ggctgggacg tgaatcggga gaagaacaat ggcgccatcc
     6001 tgtttgtgaa gaacggcctg tactatctgg gcatcatgcc aaagcagaag ggcaggtata
     6061 aggccctgag cttcgagccc acagagaaaa ccagcgaggg ctttgataag atgtactatg
     6121 actacttccc tgatgccgcc aagatgatcc caaagtgcag cacccagctg aaggccgtga
     6181 cagcccactt tcagacccac acaaccccca tcctgctgtc caacaatttc atcgagcctc
     6241 tggagatcac aaaggagatc tacgacctga acaatcctga gaaggagcca aagaagtttc
     6301 agacagccta cgccaagaaa accggcgacc agaagggcta cagagaggcc ctgtgcaagt
     6361 ggatcgactt cacaagggat tttctgtcca agtataccaa gacaacctct atcgatctgt
     6421 ctagcctgcg gccatcctct cagtataagg acctgggcga gtactatgcc gagctgaatc
     6481 ccctgctgta ccacatcagc ttccagagaa tcgccgagaa ggagatcatg gatgccgtgg
     6541 agacaggcaa gctgtacctg ttccagatct ataacaagga ctttgccaag ggccaccacg
     6601 gcaagcctaa tctgcacaca ctgtattgga ccggcctgtt ttctccagag aacctggcca
     6661 agacaagcat caagctgaat ggccaggccg agctgttcta ccgccctaag tccaggatga
     6721 agaggatggc acaccggctg ggagagaaga tgctgaacaa gaagctgaag gatcagaaaa
     6781 ccccaatccc cgacaccctg taccaggagc tgtacgacta tgtgaatcac agactgtccc
     6841 acgacctgtc tgatgaggcc agggccctgc tgcccaacgt gatcaccaag gaggtgtctc
     6901 acgagatcat caaggatagg cgctttacca gcgacaagtt ctttttccac gtgcctatca
     6961 cactgaacta tcaggccgcc aattccccat ctaagttcaa ccagagggtg aatgcctacc
     7021 tgaaggagca ccccgagaca cctatcatcg gcatcgatcg gggcgagaga aacctgatct
     7081 atatcacagt gatcgactcc accggcaaga tcctggagca gcggagcctg aacaccatcc
     7141 agcagtttga ttaccagaag aagctggaca acagggagaa ggagagggtg gcagcaaggc
     7201 aggcctggtc tgtggtgggc acaatcaagg atctgaagca gggctatctg agccaggtca
     7261 tccacgagat cgtggacctg atgatccact accaggccgt ggtggtgctg gagaacctga
     7321 atttcggctt taagagcaag aggaccggca tcgccgagaa ggccgtgtac cagcagttcg
     7381 agaagatgct gatcgataag ctgaattgcc tggtgctgaa ggactatcca gcagagaaag
     7441 tgggaggcgt gctgaaccca taccagctga cagaccagtt cacctccttt gccaagatgg
     7501 gcacccagtc tggcttcctg ttttacgtgc ctgccccata tacatctaag atcgatcccc
     7561 tgaccggctt cgtggacccc ttcgtgtgga aaaccatcaa gaatcacgag agccgcaagc
     7621 acttcctgga gggcttcgac tttctgcact acgacgtgaa aaccggcgac ttcatcctgc
     7681 actttaagat gaacagaaat ctgtccttcc agaggggcct gcccggcttt atgcctgcat
     7741 gggatatcgt gttcgagaag aacgagacac agtttgacgc caagggcacc cctttcatcg
     7801 ccggcaagag aatcgtgcca gtgatcgaga atcacagatt caccggcaga taccgggacc
     7861 tgtatcctgc caacgagctg atcgccctgc tggaggagaa gggcatcgtg ttcagggatg
     7921 gctccaacat cctgccaaag ctgctggaga atgacgattc tcacgccatc gacaccatgg
     7981 tggccctgat ccgcagcgtg ctgcagatgc ggaactccaa tgccgccaca ggcgaggact
     8041 atatcaacag ccccgtgcgc gatctgaatg gcgtgtgctt cgactcccgg tttcagaacc
     8101 cagagtggcc catggacgcc gatgccaatg gcgcctacca catcgccctg aagggccagc
     8161 tgctgctgaa tcacctgaag gagagcaagg atctgaagct gcagaacggc atctccaatc
     8221 aggactggct ggcctacatc caggagctgc gcaacgggtc caaaaggccg gcggccacga
     8281 aaaaggccgg ccaggcaaaa aagaaaaagg gatcctaccc atacgatgtt ccagattacg
     8341 cttatcccta cgacgtgcct gattatgcat acccatatga tgtccccgac tatgccggaa
     8401 gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat cctggaccga
     8461 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     8521 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     8581 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     8641 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     8701 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     8761 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     8821 cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg
     8881 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     8941 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     9001 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccggatcct
     9061 gataaattcg aaaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     9121 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     9181 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     9241 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     9301 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     9361 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     9421 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     9481 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     9541 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     9601 cttccgcgtc ctcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     9661 cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     9721 aaaagggggg actggaaggg ctaattcact cccaacgaag ataagatctg ctttttgctt
     9781 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     9841 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     9901 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     9961 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
    10021 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
    10081 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
    10141 gggggtgggg tggggcagga cagcaagggg gaggattggg aagataatag caggcatgct
    10201 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
    10261 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
    10321 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
    10381 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
    10441 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
    10501 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
    10561 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
    10621 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
    10681 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
    10741 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
    10801 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
    10861 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
    10921 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
    10981 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
    11041 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
    11101 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
    11161 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
    11221 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
    11281 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
    11341 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
    11401 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
    11461 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
    11521 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
    11581 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
    11641 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
    11701 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
    11761 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
    11821 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
    11881 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
    11941 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
    12001 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
    12061 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
    12121 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
    12181 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
    12241 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
    12301 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
    12361 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
    12421 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
    12481 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
    12541 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
    12601 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
    12661 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    12721 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    12781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    12841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    12901 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    12961 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    13021 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    13081 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
    13141 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    13201 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    13261 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    13321 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    13381 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    13441 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    13501 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    13561 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    13621 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    13681 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    13741 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    13801 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    13861 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    13921 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    13981 c
//
