LOCUS       sequence-467425-       11016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a RfxCas13d-compatible gRNA and to express HA Tagged 
            CasRx and a Puro resistance gene. It also encodes an mCherry gene. 
            The CasRx and Puro coding regions are separated by a T2A site. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11016)
  AUTHORS   Back F, Sandoval A, Vu LM, Hong VM, Bhaskara A, Rodriguez SR, 
            O'Brien JT, Kolber BJ, Kroener S, Ploski JE
  TITLE     Adeno-associated viral vector resource for the RNA-targeting Cas13d:
            A comparison of high-fidelity variants, DjCas13d and hfCas13d.
  JOURNAL   Mol Ther Methods Clin Dev. 2025 Aug 20;33(4):101565. doi: 
            10.1016/j.omtm.2025.101565. eCollection 2025 Dec 11.
  PUBMED    41020266
REFERENCE   2  (bases 1 to 11016)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther 
            Methods Clin Dev."; date: "2025-08-20"; volume: "33(4):101565. doi";
            pages: " 10.1016/j.omtm.2025.101565. eCollection 2025 Dec 11"
COMMENT     Sequence Label: pAK-DR30(SapI)-CasRx-Puro-pA/EF1A-mCherry-WPRE-pA
FEATURES             Location/Qualifiers
     source          1..11016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        651..899
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     651..671
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     822..841
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        901..930
                     /label=RfxCas13d DR30
                     /note="5' processed 30-base direct repeat for an RfxCas13d 
                     gRNA (Konermann et al., 2018)"
     enhancer        1079..1364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1757..1984
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             2008..2028
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2038..4935
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             4945..4965
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4975..5001
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5017..5070
                     /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             5071..5670
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5071..5090)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5567..5587
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5699..5716)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5705..5816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        5907..7085
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6138..7076
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7033..7053
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7098..7805
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(7240..7258)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7508..7527
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    7977..8565
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8030..8050)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8448..8459)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8796..8815)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   8929..9069
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8929..9058
                     /label=AAV2 ITR
     rep_origin      9144..9599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9231..9250)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9441..9462
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9616..9635)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9735..9757
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9795..9813)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9881..9985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9986..10054,10055..10846)
                     /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     complement(10204..10223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt gagggcctat
      661 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa
      721 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
      781 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
      841 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
      901 aacccctacc aactggtcgg ggtttgaaac ggaagagcga gctcttcttt tttttggaca
      961 gcagagatcc agtttggtta attaatgcaa agatggataa agttttaaac agagaggaat
     1021 ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattctcta gaggtacccg
     1081 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     1141 cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1201 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1261 tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta tgggactttc
     1321 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     1381 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     1441 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccggggg
     1501 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggcgcgcca
     1561 gggggggggg ggccccgaaa gtttcttttt tgtggggggg ggggcggggg ggggggcggg
     1621 ggggggggag gggcgggggg gggggaggcg gagaggtgcg gcggcggcca atcagggcgg
     1681 cgcgctccga aagtttcctt ttagggcgag gcggcggcgg cggcggccct ataaaaagcg
     1741 aagcgcgcgg cgggcgggag tcgctgcgcg ctgcctgcgc cccgtgcccc gccccgccgc
     1801 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
     1861 ggacggccct tctcctccgg gctgtaatta gctgagcaag aggtaagggt ttaagggatg
     1921 gttggttggt ggggtattaa tgtttaatta cctggagcac ctgcctgaaa tcactttttt
     1981 tcaggttgga ccggtgccac catgagcccc aagaagaaga gaaaggtgga ggccagcatc
     2041 gaaaaaaaaa agtccttcgc caagggcatg ggcgtgaagt ccacactcgt gtccggctcc
     2101 aaagtgtaca tgacaacctt cgccgaaggc agcgacgcca ggctggaaaa gatcgtggag
     2161 ggcgacagca tcaggagcgt gaatgagggc gaggccttca gcgctgaaat ggccgataaa
     2221 aacgccggct ataagatcgg caacgccaaa ttcagccatc ctaagggcta cgccgtggtg
     2281 gctaacaacc ctctgtatac aggacccgtc cagcaggata tgctcggcct gaaggaaact
     2341 ctggaaaaga ggtacttcgg cgagagcgct gatggcaatg acaatatttg tatccaggtg
     2401 atccataaca tcctggacat tgaaaaaatc ctcgccgaat acattaccaa cgccgcctac
     2461 gccgtcaaca atatctccgg cctggataag gacattattg gattcggcaa gttctccaca
     2521 gtgtatacct acgacgaatt caaagacccc gagcaccata gggccgcttt caacaataac
     2581 gataagctca tcaacgccat caaggcccag tatgacgagt tcgacaactt cctcgataac
     2641 cccagactcg gctatttcgg ccaggccttt ttcagcaagg agggcagaaa ttacatcatc
     2701 aattacggca acgaatgcta tgacattctg gccctcctga gcggactgag gcactgggtg
     2761 gtccataaca acgaagaaga gtccaggatc tccaggacct ggctctacaa cctcgataag
     2821 aacctcgaca acgaatacat ctccaccctc aactacctct acgacaggat caccaatgag
     2881 ctgaccaact ccttctccaa gaactccgcc gccaacgtga actatattgc cgaaactctg
     2941 ggaatcaacc ctgccgaatt cgccgaacaa tatttcagat tcagcattat gaaagagcag
     3001 aaaaacctcg gattcaatat caccaagctc agggaagtga tgctggacag gaaggatatg
     3061 tccgagatca ggaaaaatca taaggtgttc gactccatca ggaccaaggt ctacaccatg
     3121 atggactttg tgatttatag gtattacatc gaagaggatg ccaaggtggc tgccgccaat
     3181 aagtccctcc ccgataatga gaagtccctg agcgagaagg atatctttgt gattaacctg
     3241 aggggctcct tcaacgacga ccagaaggat gccctctact acgatgaagc taatagaatt
     3301 tggagaaagc tcgaaaatat catgcacaac atcaaggaat ttaggggaaa caagacaaga
     3361 gagtataaga agaaggacgc ccctagactg cccagaatcc tgcccgctgg ccgtgatgtt
     3421 tccgccttca gcaaactcat gtatgccctg accatgttcc tggatggcaa ggagatcaac
     3481 gacctcctga ccaccctgat taataaattc gataacatcc agagcttcct gaaggtgatg
     3541 cctctcatcg gagtcaacgc taagttcgtg gaggaatacg cctttttcaa agactccgcc
     3601 aagatcgccg atgagctgag gctgatcaag tccttcgcta gaatgggaga acctattgcc
     3661 gatgccagga gggccatgta tatcgacgcc atccgtattt taggaaccaa cctgtcctat
     3721 gatgagctca aggccctcgc cgacaccttt tccctggacg agaacggaaa caagctcaag
     3781 aaaggcaagc acggcatgag aaatttcatt attaataacg tgatcagcaa taaaaggttc
     3841 cactacctga tcagatacgg tgatcctgcc cacctccatg agatcgccaa aaacgaggcc
     3901 gtggtgaagt tcgtgctcgg caggatcgct gacatccaga aaaaacaggg ccagaacggc
     3961 aagaaccaga tcgacaggta ctacgaaact tgtatcggaa aggataaggg caagagcgtg
     4021 agcgaaaagg tggacgctct cacaaagatc atcaccggaa tgaactacga ccaattcgac
     4081 aagaaaagga gcgtcattga ggacaccggc agggaaaacg ccgagaggga gaagtttaaa
     4141 aagatcatca gcctgtacct caccgtgatc taccacatcc tcaagaatat tgtcaatatc
     4201 aacgccaggt acgtcatcgg attccattgc gtcgagcgtg atgctcaact gtacaaggag
     4261 aaaggctacg acatcaatct caagaaactg gaagagaagg gattcagctc cgtcaccaag
     4321 ctctgcgctg gcattgatga aactgccccc gataagagaa aggacgtgga aaaggagatg
     4381 gctgaaagag ccaaggagag cattgacagc ctcgagagcg ccaaccccaa gctgtatgcc
     4441 aattacatca aatacagcga cgagaagaaa gccgaggagt tcaccaggca gattaacagg
     4501 gagaaggcca aaaccgccct gaacgcctac ctgaggaaca ccaagtggaa tgtgatcatc
     4561 agggaggacc tcctgagaat tgacaacaag acatgtaccc tgttcagaaa caaggccgtc
     4621 cacctggaag tggccaggta tgtccacgcc tatatcaacg acattgccga ggtcaattcc
     4681 tacttccaac tgtaccatta catcatgcag agaattatca tgaatgagag gtacgagaaa
     4741 agcagcggaa aggtgtccga gtacttcgac gctgtgaatg acgagaagaa gtacaacgat
     4801 aggctcctga aactgctgtg tgtgcctttc ggctactgta tccccaggtt taagaacctg
     4861 agcatcgagg ccctgttcga taggaacgag gccgccaagt tcgacaagga gaaaaagaag
     4921 gtgtccggca attccggatc cggacctaag aaaaagagga aggtggcggc cgcttaccca
     4981 tacgatgttc cagattacgc tgctagcggc agtggagagg gcagaggaag tctgctaaca
     5041 tgcggtgacg tcgaggagaa tcctggccca atgaccgagt acaagcccac ggtgcgcctc
     5101 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5161 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5221 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5281 gccgccgtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5341 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5401 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc
     5461 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5521 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5581 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5641 tggtgcatga cccgcaagcc cggtgcctga cgtacgctaa ctagagctcg ctgatcagcc
     5701 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     5761 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     5821 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     5881 gattgggaag agaatagcag gcatgcggct ccggtgcccg tcagtgggca gagcgcacat
     5941 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     6001 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     6061 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     6121 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     6181 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     6241 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     6301 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     6361 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     6421 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     6481 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     6541 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     6601 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     6661 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     6721 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     6781 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     6841 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     6901 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     6961 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     7021 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     7081 cgtgaggtac cggatccgtg agcaagggcg aggaggataa catggccatc atcaaggagt
     7141 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     7201 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     7261 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     7321 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     7381 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     7441 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     7501 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     7561 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     7621 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     7681 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     7741 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     7801 agtaaggcgc gcccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     7861 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     7921 aacctggccc tgtcttcttg acgagcattc ctaaattcga tatcaagctt atcgataatc
     7981 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     8041 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     8101 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     8161 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     8221 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     8281 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     8341 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     8401 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     8461 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     8521 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     8581 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     8641 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     8701 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     8761 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     8821 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     8881 tcagcctccc gagttgttgg gattccaggc atgctgggga gcggccgcag gaacccctag
     8941 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     9001 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     9061 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     9121 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     9181 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     9241 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     9301 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     9361 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     9421 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     9481 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     9541 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     9601 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     9661 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     9721 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     9781 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     9841 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     9901 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     9961 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
    10021 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
    10081 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
    10141 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
    10201 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
    10261 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
    10321 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
    10381 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
    10441 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
    10501 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
    10561 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
    10621 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
    10681 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
    10741 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
    10801 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
    10861 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
    10921 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
    10981 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
