LOCUS       sequence-457833-       12672 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hfCas13d (N2V8) in mammalian cells for knockdown of target
            RNAs in combination with compatible crRNAs. Localized to the 
            nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12672)
  AUTHORS   Hart SK, Wessels HH, Mendez-Mancilla A, Muller S, Drabavicius G, 
            Choi O, Sanjana NE
  TITLE     Low copy CRISPR-Cas13d mitigates collateral RNA cleavage.
  JOURNAL   bioRxiv [Preprint]. 2024 May 14:2024.05.13.594039. doi: 
            10.1101/2024.05.13.594039.
  PUBMED    38798586
REFERENCE   2  (bases 1 to 12672)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 May 14:2024.05.13.594039. doi: 
            10.1101/2024.05.13.594039."
COMMENT     Sequence Label: 
            pLentiRNACRISPR_008-TetO-NLS-hfCas13d-NLS-WPRE-EFS-rtTA3-2A-Blast
FEATURES             Location/Qualifiers
     source          1..12672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2904..2924
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2934..2960
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3010..3598
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3063..3083)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3481..3492)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3601..3617)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3602..3618)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        3663..3874
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3899..4603
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     CDS             4613..4669
                     /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             4670..5068
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5460..5640
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5666..5683)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5672..5896
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5942..6370
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6029..6048)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6239..6260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6379..6399)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6384..6713
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6564..6699
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6626..6645
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6761..6808
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6827..7201
                     /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    7331..7452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7368..7387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7422..7441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7501..7517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7501..7517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7514..7536)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7525..7541
                     /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(7549..7555,7556..7573,7574..7579))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7594..7615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7732..7749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7903..8491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7983..8002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8662..9453,9454..9522))
                     /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     9285..9304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9523..9627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9702..9721)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9893..10272
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10274..10472
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             10490..10670
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10717..10842
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11335..11568
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11753..11797
                     /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    12095..12212
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    12257..12277
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    12322..12340
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12357..12375
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12393..12411
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12429..12447
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12464..12482
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12500..12518
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        12530..12568
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             12633..12653
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(12669..12672,1..2894)
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILVEYITNVVYVVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
ORIGIN
        1 aaaaaaaaaa gtccttcgcc aagggcatgg gcgtgaagtc cacactcgtg tccggctcca
       61 aagtgtacat gacaaccttc gccgaaggca gcgacgccag gctggaaaag atcgtggagg
      121 gcgacagcat caggagcgtg aatgagggcg aggccttcag cgctgaaatg gccgataaaa
      181 acgccggcta taagatcggc aacgccaaat tcagccatcc taagggctac gccgtggtgg
      241 ctaacaaccc tctgtataca ggacccgtcc agcaggatat gctcggcctg aaggaaactc
      301 tggaaaagag gtacttcggc gagagcgctg atggcaatga caatatttgt atccaggtga
      361 tccataacat cctggacatt gagaagatcc tggtggagta catcacaaac gtggtgtacg
      421 tggtgaacaa catctccggc ctggataagg acattattgg attcggcaag ttctccacag
      481 tgtataccta cgacgaattt aaagaccccg agcaccatag ggccgctttc aacaataacg
      541 ataagctcat caacgccatc aaggcccagt atgacgagtt cgacaacttc ctcgataacc
      601 ccagactcgg ctatttcggc caggcctttt tcagcaagga gggcagaaat tacatcatca
      661 attacggcaa cgaatgctat gacattctgg ccctcctgag cggactgagg cactgggtgg
      721 tccataacaa cgaagaagag tccaggatct ccaggacctg gctctacaac ctcgataaga
      781 acctcgacaa cgaatacatc tccaccctca actacctcta cgacaggatc accaatgagc
      841 tgaccaactc cttctccaag aactccgccg ccaacgtgaa ctatattgcc gaaactctgg
      901 gaatcaaccc tgccgaattt gccgaacaat atttcagatt cagcattatg aaagagcaga
      961 aaaacctcgg attcaatatc accaagctca gggaagtgat gctggacagg aaggatatgt
     1021 ccgagatcag gaaaaatcat aaggtgttcg actccatcag gaccaaggtc tacaccatga
     1081 tggactttgt gatttatagg tattacatcg aagaggatgc caaggtggct gccgccaata
     1141 agtccctccc cgataatgag aagtccctga gcgagaagga tatctttgtg attaacctga
     1201 ggggctcctt caacgacgac cagaaggatg ccctctacta cgatgaagct aatagaattt
     1261 ggagaaagct cgaaaatatc atgcacaaca tcaaggaatt taggggaaac aagacaagag
     1321 agtataagaa gaaggacgcc cctagactgc ccagaatcct gcccgctggc cgtgatgttt
     1381 ccgccttcag caaactcatg tatgccctga ccatgttcct ggatggcaag gagatcaacg
     1441 acctcctgac caccctgatt aataaattcg ataacatcca gagcttcctg aaggtgatgc
     1501 ctctcatcgg agtcaacgct aagttcgtgg aggaatacgc ctttttcaaa gactccgcca
     1561 agatcgccga tgagctgagg ctgatcaagt ccttcgctag aatgggagaa cctattgccg
     1621 atgccaggag ggccatgtat atcgacgcca tccgtatttt aggaaccaac ctgtcctatg
     1681 atgagctcaa ggccctcgcc gacacctttt ccctggacga gaacggaaac aagctcaaga
     1741 aaggcaagca cggcatgaga aatttcatta ttaataacgt gatcagcaat aaaaggttcc
     1801 actacctgat cagatacggt gatcctgccc acctccatga gatcgccaaa aacgaggccg
     1861 tggtgaagtt cgtgctcggc aggatcgctg acatccagaa aaaacagggc cagaacggca
     1921 agaaccagat cgacaggtac tacgaaactt gtatcggaaa ggataagggc aagagcgtga
     1981 gcgaaaaggt ggacgctctc acaaagatca tcaccggaat gaactacgac caattcgaca
     2041 agaaaaggag cgtcattgag gacaccggca gggaaaacgc cgagagggag aagtttaaaa
     2101 agatcatcag cctgtacctc accgtgatct accacatcct caagaatatt gtcaatatca
     2161 acgccaggta cgtcatcgga ttccattgcg tcgagcgtga tgctcaactg tacaaggaga
     2221 aaggctacga catcaatctc aagaaactgg aagagaaggg attcagctcc gtcaccaagc
     2281 tctgcgctgg cattgatgaa actgcccccg ataagagaaa ggacgtggaa aaggagatgg
     2341 ctgaaagagc caaggagagc attgacagcc tcgagagcgc caaccccaag ctgtatgcca
     2401 attacatcaa atacagcgac gagaagaaag ccgaggagtt caccaggcag attaacaggg
     2461 agaaggccaa aaccgccctg aacgcctacc tgaggaacac caagtggaat gtgatcatca
     2521 gggaggacct cctgagaatt gacaacaaga catgtaccct gttcagaaac aaggccgtcc
     2581 acctggaagt ggccaggtat gtccacgcct atatcaacga cattgccgag gtcaattcct
     2641 acttccaact gtaccattac atcatgcaga gaattatcat gaatgagagg tacgagaaaa
     2701 gcagcggaaa ggtgtccgag tacttcgacg ctgtgaatga cgagaagaag tacaacgata
     2761 ggctcctgaa actgctgtgt gtgcctttcg gctactgtat ccccaggttt aagaacctga
     2821 gcatcgaggc cctgttcgat aggaacgagg ccgccaagtt cgacaaggag aaaaagaagg
     2881 tgtccggcaa ttccggatcc ggacctaaga aaaagaggaa ggtggcggcc gcttacccat
     2941 acgatgttcc agattacgct taaggtacaa agtggtgaat tccggcaatt cgatatcaag
     3001 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3061 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3121 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3181 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3241 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3301 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3361 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3421 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3481 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3541 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3601 cgataccgtc gacctcgagt aggtcttgaa aggagtggga attggctccg gtgcccgtca
     3661 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     3721 atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     3781 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     3841 tctttttcgc aacgggtttg ccgccagaac acaggaccgg ttctagagcg ctgccaccat
     3901 gtctaggctg gacaagagca aagtcataaa cggagctctg gaattactca atggtgtcgg
     3961 tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct
     4021 gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga
     4081 caggcatcat acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa
     4141 caacgccaag tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct
     4201 cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg
     4261 tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac
     4321 actgggctgc gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc
     4381 taccaccgat tctatgcccc cacttctgag acaagcaatt gagctgttcg accggcaggg
     4441 agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct
     4501 aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc
     4561 agccgatgcc cttgacgatt ttgaccttga catgctcccc gggggatccg gcgcaacaaa
     4621 cttctctctg ctgaaacaag ccggagatgt cgaagagaat cctggaccga tggccaagcc
     4681 tttgtctcaa gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc
     4741 catctctgaa gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac
     4801 tggtgtcaat gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac
     4861 tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag
     4921 gggcatcttg agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat
     4981 caaagccata gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct
     5041 gccctctggt tatgtgtggg agggctaagg tacctttaag accaatgact tacaaggcag
     5101 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     5161 aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     5221 ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca
     5281 agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct
     5341 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga
     5401 ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg
     5461 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     5521 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     5581 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     5641 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     5701 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     5761 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     5821 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     5881 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     5941 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     6001 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     6061 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     6121 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     6181 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     6241 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     6301 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6361 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     6421 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     6481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     6541 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     6601 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     6661 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     6721 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     6781 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
     6841 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
     6901 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
     6961 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
     7021 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
     7081 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
     7141 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
     7201 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     7261 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     7321 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7381 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7441 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     7501 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7561 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7621 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7681 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7741 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7801 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7861 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7921 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7981 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     8041 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     8101 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     8161 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     8221 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     8281 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     8341 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     8401 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     8461 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     8521 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8581 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8641 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8701 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8761 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8821 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8881 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8941 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     9001 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     9061 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     9121 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     9181 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     9241 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     9301 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     9361 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     9421 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     9481 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9541 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9601 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     9661 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     9721 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     9781 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     9841 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     9901 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     9961 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
    10021 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
    10081 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
    10141 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
    10201 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
    10261 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
    10321 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
    10381 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
    10441 aggcgtgtac ggtgggaggt ctatataagc agcgcgtttt gcctgtactg ggtctctctg
    10501 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    10561 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    10621 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
    10681 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
    10741 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
    10801 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
    10861 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
    10921 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
    10981 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
    11041 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
    11101 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
    11161 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
    11221 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
    11281 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
    11341 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
    11401 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
    11461 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
    11521 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
    11581 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
    11641 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
    11701 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
    11761 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
    11821 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
    11881 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
    11941 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
    12001 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc
    12061 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
    12121 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
    12181 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
    12241 cagtttggtt aattaacaac tttgtataga aaagttggct ccgaattcgc ccttcaggtc
    12301 cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc
    12361 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
    12421 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga
    12481 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt
    12541 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagattg caaagggcga
    12601 attcgaccca agtttgtaca gccaccatga gccccaagaa gaagagaaag gtggaggcca
    12661 gcgaattcat cg
//
