LOCUS       sequence-266643-       12358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PguCas13b in mammalian cells, localized in the cytoplasm. 
            For cloning of guide RNAs compatible with PguCas13b. Contains a 3' 
            direct repeat. Clone using BsmBI. F overhang cacc. R overhang caac.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12358)
  AUTHORS   Wessels HH, Mendez-Mancilla A, Guo X, Legut M, Daniloski Z, Sanjana 
            NE
  TITLE     Massively parallel Cas13 screens reveal principles for guide RNA 
            design.
  JOURNAL   Nat Biotechnol. 2020 Jun;38(6):722-727. doi: 
            10.1038/s41587-020-0456-9. Epub 2020 Mar 16.
  PUBMED    32518401
REFERENCE   2  (bases 1 to 12358)
  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: 
            "10.1038/s41587-020-0456-9"; journalName: "Nat Biotechnol"; date: 
            "2020-06"; volume: "38"; issue: "6"; pages: "722-727"
COMMENT     Sequence Label: pLentiRNACRISPR_002 -
            hU6-DR_BsmBI-EFS-PguCas1...
FEATURES             Location/Qualifiers
     source          1..12358
                     /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        2608..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             complement(2878..2889)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2969..3180
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             6736..6762
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             6781..6837
                     /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             6838..7434
                     /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     7331..7351
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7450..8038
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7503..7523)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7921..7932)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8110..8343
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8369..8386)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8375..8599
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8645..9073
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8732..8751)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8942..8963
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9082..9102)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9087..9416
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9267..9402
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9329..9348
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9464..9511
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9530..9904
                     /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    10034..10155
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10071..10090)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10125..10144
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10204..10220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10204..10220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10217..10239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10228..10244
                     /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(10252..10258,10259..10276,10277..10282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10297..10318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10435..10452)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10606..11194)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10686..10705)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11365..12156,12157..12225))
                     /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     11988..12007
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12226..12330)
                     /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 aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccgaga cggacgtctc tgttggatct
     2881 accctctatt tgaagggtac acacaacttt tttgaattcg ctagctaggt cttgaaagga
     2941 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     3001 aagttggggg gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac
     3061 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     3121 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     3181 gaccggttct agagcgctgc caccatgaca gagcagagcg agaggcccta caacggcacc
     3241 tactacaccc tggaagataa gcacttctgg gccgcctttc tgaacctggc cagacacaac
     3301 gcctacatca ccctgacaca catcgaccgg cagctggcct acagcaaggc cgacatcacc
     3361 aacgaccagg acgtgctgag cttcaaggcc ctgtggaaga acttcgacaa cgacctggaa
     3421 agaaagagcc ggctgcggag cctgatcctg aagcacttca gctttctgga aggcgccgct
     3481 tacggcaaga agctgttcga gtctaagagc agcggcaaca agagcagcaa gaacaaagag
     3541 ctgaccaaga aagagaaaga ggaactgcag gccaacgctc tgagcctgga caacctgaag
     3601 tctatcctgt tcgacttcct gcagaagctg aaggacttcc ggaactacta cagccactac
     3661 cggcacagcg gaagcagcga gctgcctctg ttcgacggca acatgctgca gcggctgtac
     3721 aacgtgttcg acgtgtccgt gcagcgcgtg aagatcgacc acgagcacaa cgatgaggtg
     3781 gaccctcact accacttcaa ccacctcgtg cggaagggca agaaggacag atacggccac
     3841 aacgacaacc ccagcttcaa gcaccacttc gtggacggcg aaggcatggt tacagaagcc
     3901 ggcctgctgt tcttcgtgtc cctgttcctg gaaaagcggg acgccatctg gatgcagaag
     3961 aagatccggg gcttcaaagg cggcaccgag acataccagc agatgaccaa cgaggtgttc
     4021 tgccggtcca gaatcagcct gcctaagctg aagctggaaa gcctgcggat ggacgactgg
     4081 atgctgctgg acatgctgaa cgaactcgtg cggtgcccca agcctctgta cgacagactg
     4141 agagaggacg accgggcctg ctttagagtg cccgtggata tcctgcctga cgaggacgat
     4201 acagatggcg gcggagagga ccccttcaag aataccctcg tccggcacca ggacagattc
     4261 ccttacttcg ccctgcggta cttcgacctg aagaaggtgt tcaccagcct gcggttccac
     4321 atcgatctgg gcacctacca ctttgccatc tacaagaaga tgatcggcga gcagcccgag
     4381 gacagacacc tgaccagaaa cctgtacggc ttcggccgga tccaggactt cgccgaagaa
     4441 cacagacccg aggaatggaa gcggctcgtc agagatctgg actacttcga gacaggcgac
     4501 aagccctaca tcagccagac aagcccacac taccacatcg agaagggaaa gatcggcctg
     4561 agattcatgc ccgagggcca gcatctttgg ccctctccag aagtgggcac caccagaacc
     4621 ggcagatcta agtacgccca ggacaagaga ctgaccgccg aggcctttct gtctgtgcac
     4681 gagctgatgc ctatgatgtt ctactacttc ctgctgcgcg agaagtacag cgaggaagtg
     4741 tctgccgaga gagtgcaggg cagaatcaag cgcgtgatcg aggatgtgta cgccgtgtac
     4801 gatgcctttg ccagggacga gatcaacacc cgggatgagc tggatgcctg cctggccgat
     4861 aagggcatca gaagaggcca tctgcctcgc cagatgatcg ccatcctgag ccaagagcac
     4921 aaggacatgg aagagaagat tcggaagaaa ctgcaagaga tgatggccga caccgaccac
     4981 cggctggata tgctggatag acagaccgac cggaagatca gaatcggccg gaagaatgcc
     5041 ggactgccta agagcggagt gatcgccgat tggctcgtgc gggacatgat gagatttcag
     5101 cccgtggcca aggacgccag cggaaagcct ctgaacaact ccaaggccaa cagcaccgag
     5161 taccggatgc tgcagagagc cctggctctg tttggaggcg agaaagagag gctgacccca
     5221 tacttccggc agatgaatct gaccggcgga aacaaccctc atccttttct gcacgaaact
     5281 cgctgggaga gccacaccaa catcctgtcc ttttaccggt cctacctgag agcccgcaag
     5341 gccttcctcg aaaggatcgg cagatccgac agagtggaaa accggccatt tctgctgctg
     5401 aaagagccca agaccgacag acagacactg gtggctggat ggaagggcga gttccatctg
     5461 ccaagaggca tcttcacaga ggccgtgcgg gattgcctga tcgagatggg acacgatgaa
     5521 gtggccagct acaaagaagt gggattcatg gccaaggccg tgccactgta ctttgagaga
     5581 gcctgcgagg acagggtgca gcccttttac gacagcccct tcaacgtggg caacagcctg
     5641 aagcctaaga agggcagatt cctgagcaaa gaggaacggg ccgaagagtg ggagcggggc
     5701 aaagagagat tccgggatct cgaagcctgg tcctatagcg ccgccagaag aatcgaggac
     5761 gcctttgccg gcatcgagta tgctagcccc ggaaacaaga agaaaatcga gcagctcctg
     5821 cgggacctga gcctgtggga agcctttgag agcaagctga aagtgcgggc cgaccggatc
     5881 aatctggcca agctgaagaa agaaatcctg gaagcccaag aacaccccta ccacgacttc
     5941 aagagctggc agaagttcga gcgcgagctg cggctggtca agaaccagga tatcatcacc
     6001 tggatgatgt gccgcgacct gatggaagaa aacaaggtgg aaggcctgga caccggcaca
     6061 ctgtacctga aggacatcag acccaacgtg caagagcagg gcagcctgaa cgtgctgaac
     6121 agagtgaagc ccatgagact gcccgtggtg gtgtacagag ccgatagcag aggccacgtg
     6181 cacaaagaag aagcccctct ggccaccgtg tacatcgagg aaagagacac caagctgctg
     6241 aagcagggca acttcaagtc cttcgtgaag gacaggcggc tgaacggcct gttctccttc
     6301 gttgatactg gcggactggc catggaacag taccccatct ccaagctgag agtggaatac
     6361 gagctggcca agtaccagac cgccagagtg tgtgtgttcg agctgaccct gagactggaa
     6421 gagtccctgc tgacccggta tcctcatctg cccgacgaga gcttcagaga gatgctggaa
     6481 tcttggagcg accctctgct ggccaaatgg cctgaactgc acggaaaagt gcggctgctg
     6541 atcgccgtgc ggaatgcctt tagccacaat cagtacccta tgtacgacga ggccgtgttc
     6601 agctccatcc ggaagtacga tcctagcagc cccgacgcca tcgaagagag aatgggcctg
     6661 aatatcgccc accgcctgtc tgaggaagtg aagcaggcca aagaaaccgt ggaacggatc
     6721 attcaggccc ttcaactgcc tccacttgaa agactgacac tgggtagttc cggatccggc
     6781 gcaacaaact tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccgacc
     6841 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     6901 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     6961 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     7021 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     7081 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     7141 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     7201 tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     7261 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     7321 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     7381 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaacgcgt
     7441 taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     7501 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     7561 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     7621 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     7681 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     7741 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     7801 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     7861 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     7921 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     7981 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
     8041 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     8101 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     8161 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     8221 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     8281 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     8341 gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     8401 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     8461 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     8521 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     8581 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     8641 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     8701 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     8761 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     8821 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     8881 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     8941 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     9001 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     9061 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     9121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     9181 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     9241 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     9301 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     9361 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     9421 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     9481 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
     9541 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
     9601 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
     9661 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
     9721 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
     9781 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
     9841 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
     9901 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     9961 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
    10021 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
    10081 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    10141 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
    10201 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
    10261 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
    10321 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
    10381 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
    10441 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
    10501 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
    10561 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
    10621 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
    10681 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
    10741 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
    10801 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
    10861 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
    10921 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
    10981 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
    11041 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
    11101 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
    11161 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
    11221 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
    11281 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
    11341 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
    11401 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
    11461 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
    11521 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
    11581 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
    11641 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
    11701 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
    11761 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
    11821 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
    11881 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
    11941 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    12001 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    12061 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    12121 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    12181 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    12241 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    12301 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
