LOCUS       sequence-456689-       12666 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses inactive RfxCas13d in mammalian cells for binding 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 12666)
  TITLE     Low copy CRISPR-Cas13d mitigates collateral RNA cleavage
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12666)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: 
            pLentiRNACRISPR_011-TetO-NLS-inactive-RfxCas13d-NLS-WPRE-EFS-rtTA3-2
            A-Blast
FEATURES             Location/Qualifiers
     source          1..12666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             275..331
                     /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             332..730
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             1122..1302
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(1328..1345)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1334..1558
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1604..2032
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1691..1710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1901..1922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2041..2061)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2046..2375
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2226..2361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2288..2307
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2423..2470
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2489..2863
                     /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    2993..3114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3030..3049)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3084..3103
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3163..3179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3163..3179)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3176..3198)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3187..3203
                     /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(3211..3217,3218..3235,3236..3241))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3256..3277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3394..3411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3565..4153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3645..3664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4324..5115,5116..5184))
                     /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     4947..4966
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5185..5289)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5364..5383)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5555..5934
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5936..6134
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6152..6332
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6379..6504
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6997..7230
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7415..7459
                     /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    7757..7874
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    7919..7939
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    7984..8002
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8019..8037
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8055..8073
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8091..8109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8126..8144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8162..8180
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        8192..8230
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8227..8251
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             8295..8315
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8325..11222
                     /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
                     YGNECYDILALLSGLAHWVVANNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFANKAVALEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             11232..11252
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11262..11288
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    11338..11926
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11391..11411)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11809..11820)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11929..11945)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11930..11946)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        11991..12202
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(12227..12666,1..265)
                     /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"
ORIGIN
        1 ggaacaggag catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc
       61 cccacttctg agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct
      121 tttcggcctg gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg
      181 gccgaccgac gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga
      241 ttttgacctt gacatgctcc ccgggggatc cggcgcaaca aacttctctc tgctgaaaca
      301 agccggagat gtcgaagaga atcctggacc gatggccaag cctttgtctc aagaagaatc
      361 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
      421 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
      481 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
      541 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
      601 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
      661 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
      721 ggagggctaa ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
      781 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc
      841 ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag
      901 ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
      961 aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc
     1021 tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag
     1081 catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc
     1141 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     1201 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     1261 gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc
     1321 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1381 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1441 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1501 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     1561 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     1621 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1681 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1741 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1801 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1861 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1921 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1981 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2041 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2101 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2161 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2221 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2281 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     2341 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     2401 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     2461 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct
     2521 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
     2581 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
     2641 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
     2701 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
     2761 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
     2821 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
     2881 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     2941 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     3001 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3061 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     3121 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     3181 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3241 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3301 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3361 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3421 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3481 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3541 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3601 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3661 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3721 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3781 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3841 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3901 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3961 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4021 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4081 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4141 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4201 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4261 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4321 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4381 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4441 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4501 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4561 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4621 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4681 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4741 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4801 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4861 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4921 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4981 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5041 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5101 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5161 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5221 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5281 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     5341 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     5401 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     5461 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     5521 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     5581 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     5641 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     5701 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5761 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     5821 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5881 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     5941 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     6001 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     6061 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     6121 gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc agatctgagc
     6181 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6241 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6301 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
     6361 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
     6421 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     6481 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
     6541 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
     6601 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     6661 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     6721 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     6781 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     6841 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     6901 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     6961 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     7021 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     7081 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     7141 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     7201 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     7261 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     7321 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     7381 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     7441 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     7501 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     7561 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     7621 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     7681 cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg
     7741 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     7801 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     7861 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaaca
     7921 actttgtata gaaaagttgg ctccgaattc gcccttcagg tccgaggttc tagacgagtt
     7981 tactccctat cagtgataga gaacgatgtc gagtttactc cctatcagtg atagagaacg
     8041 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
     8101 tgatagagaa cgtatgtcga gtttatccct atcagtgata gagaacgtat gtcgagttta
     8161 ctccctatca gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg aggcctatat
     8221 aagcagagct cgtttagtga accgtcagat cgcaaagggc gaattcgacc caagtttgta
     8281 cagccaccat gagccccaag aagaagagaa aggtggaggc cagcatcgaa aaaaaaaagt
     8341 ccttcgccaa gggcatgggc gtgaagtcca cactcgtgtc cggctccaaa gtgtacatga
     8401 caaccttcgc cgaaggcagc gacgccaggc tggaaaagat cgtggagggc gacagcatca
     8461 ggagcgtgaa tgagggcgag gccttcagcg ctgaaatggc cgataaaaac gccggctata
     8521 agatcggcaa cgccaaattc agccatccta agggctacgc cgtggtggct aacaaccctc
     8581 tgtatacagg acccgtccag caggatatgc tcggcctgaa ggaaactctg gaaaagaggt
     8641 acttcggcga gagcgctgat ggcaatgaca atatttgtat ccaggtgatc cataacatcc
     8701 tggacattga aaaaatcctc gccgaataca ttaccaacgc cgcctacgcc gtcaacaata
     8761 tctccggcct ggataaggac attattggat tcggcaagtt ctccacagtg tatacctacg
     8821 acgaattcaa agaccccgag caccataggg ccgctttcaa caataacgat aagctcatca
     8881 acgccatcaa ggcccagtat gacgagttcg acaacttcct cgataacccc agactcggct
     8941 atttcggcca ggcctttttc agcaaggagg gcagaaatta catcatcaat tacggcaacg
     9001 aatgctatga cattctggcc ctcctgagcg gactggcgca ctgggtggtc gctaacaacg
     9061 aagaagagtc caggatctcc aggacctggc tctacaacct cgataagaac ctcgacaacg
     9121 aatacatctc caccctcaac tacctctacg acaggatcac caatgagctg accaactcct
     9181 tctccaagaa ctccgccgcc aacgtgaact atattgccga aactctggga atcaaccctg
     9241 ccgaattcgc cgaacaatat ttcagattca gcattatgaa agagcagaaa aacctcggat
     9301 tcaatatcac caagctcagg gaagtgatgc tggacaggaa ggatatgtcc gagatcagga
     9361 aaaatcataa ggtgttcgac tccatcagga ccaaggtcta caccatgatg gactttgtga
     9421 tttataggta ttacatcgaa gaggatgcca aggtggctgc cgccaataag tccctccccg
     9481 ataatgagaa gtccctgagc gagaaggata tctttgtgat taacctgagg ggctccttca
     9541 acgacgacca gaaggatgcc ctctactacg atgaagctaa tagaatttgg agaaagctcg
     9601 aaaatatcat gcacaacatc aaggaattta ggggaaacaa gacaagagag tataagaaga
     9661 aggacgcccc tagactgccc agaatcctgc ccgctggccg tgatgtttcc gccttcagca
     9721 aactcatgta tgccctgacc atgttcctgg atggcaagga gatcaacgac ctcctgacca
     9781 ccctgattaa taaattcgat aacatccaga gcttcctgaa ggtgatgcct ctcatcggag
     9841 tcaacgctaa gttcgtggag gaatacgcct ttttcaaaga ctccgccaag atcgccgatg
     9901 agctgaggct gatcaagtcc ttcgctagaa tgggagaacc tattgccgat gccaggaggg
     9961 ccatgtatat cgacgccatc cgtattttag gaaccaacct gtcctatgat gagctcaagg
    10021 ccctcgccga caccttttcc ctggacgaga acggaaacaa gctcaagaaa ggcaagcacg
    10081 gcatgagaaa tttcattatt aataacgtga tcagcaataa aaggttccac tacctgatca
    10141 gatacggtga tcctgcccac ctccatgaga tcgccaaaaa cgaggccgtg gtgaagttcg
    10201 tgctcggcag gatcgctgac atccagaaaa aacagggcca gaacggcaag aaccagatcg
    10261 acaggtacta cgaaacttgt atcggaaagg ataagggcaa gagcgtgagc gaaaaggtgg
    10321 acgctctcac aaagatcatc accggaatga actacgacca attcgacaag aaaaggagcg
    10381 tcattgagga caccggcagg gaaaacgccg agagggagaa gtttaaaaag atcatcagcc
    10441 tgtacctcac cgtgatctac cacatcctca agaatattgt caatatcaac gccaggtacg
    10501 tcatcggatt ccattgcgtc gagcgtgatg ctcaactgta caaggagaaa ggctacgaca
    10561 tcaatctcaa gaaactggaa gagaagggat tcagctccgt caccaagctc tgcgctggca
    10621 ttgatgaaac tgcccccgat aagagaaagg acgtggaaaa ggagatggct gaaagagcca
    10681 aggagagcat tgacagcctc gagagcgcca accccaagct gtatgccaat tacatcaaat
    10741 acagcgacga gaagaaagcc gaggagttca ccaggcagat taacagggag aaggccaaaa
    10801 ccgccctgaa cgcctacctg aggaacacca agtggaatgt gatcatcagg gaggacctcc
    10861 tgagaattga caacaagaca tgtaccctgt tcgcaaacaa ggccgtcgcc ctggaagtgg
    10921 ccaggtatgt ccacgcctat atcaacgaca ttgccgaggt caattcctac ttccaactgt
    10981 accattacat catgcagaga attatcatga atgagaggta cgagaaaagc agcggaaagg
    11041 tgtccgagta cttcgacgct gtgaatgacg agaagaagta caacgatagg ctcctgaaac
    11101 tgctgtgtgt gcctttcggc tactgtatcc ccaggtttaa gaacctgagc atcgaggccc
    11161 tgttcgatag gaacgaggcc gccaagttcg acaaggagaa aaagaaggtg tccggcaatt
    11221 ccggatccgg acctaagaaa aagaggaagg tggcggccgc ttacccatac gatgttccag
    11281 attacgctta acgtacaaag tggtgaattc cggcaattcg atatcaagct tatcgataat
    11341 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
    11401 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
    11461 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
    11521 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
    11581 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
    11641 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
    11701 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
    11761 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
    11821 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
    11881 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
    11941 cctcgagtag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
    12001 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta
    12061 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
    12121 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
    12181 cgggtttgcc gccagaacac aggaccggtt ctagagcgct gccaccatgt ctaggctgga
    12241 caagagcaaa gtcataaacg gagctctgga attactcaat ggtgtcggta tcgaaggcct
    12301 gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt
    12361 gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac
    12421 ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc
    12481 ataccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc
    12541 aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt
    12601 ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt
    12661 attgga
//
