LOCUS       sequence-456687-       12672 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses inactive hfCas13d 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 12672)
  TITLE     Low copy CRISPR-Cas13d mitigates collateral RNA cleavage
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12672)
  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            "Unpublished"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_012-TetO-NLS-inactive-hfCas13d-NLS-WPRE-EFS-rtTA3-2A
            -Blast
FEATURES             Location/Qualifiers
     source          1..12672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    221..454
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    947..1072
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1119..1299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1317..1515)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        1517..1896
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2068..2087
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2162..2266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2267..2335,2336..3127)
                     /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(2485..2504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3298..3886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3787..3806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4040..4057
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4174..4195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4210..4215,4216..4233,4234..4240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4248..4264
                     /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)."
     primer_bind     4253..4275
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4272..4288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4272..4288
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4337..4458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4348..4367)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4402..4421
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4588..4962)
                     /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"
     promoter        complement(4981..5028)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5076..5405)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5090..5225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5144..5163)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5390..5410
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5419..5847)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5529..5550)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5741..5760
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5893..6117
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6106..6123
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6149..6329
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     CDS             complement(6721..7119)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(7120..7176)
                     /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             complement(7186..7890)
                     /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"
     promoter        complement(7915..8126)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     8171..8187
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8172..8188
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8191..8779
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             8297..8308
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     8706..8726
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8829..8855)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(8865..8885)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8895..11792)
                     /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
                     YGNECYDILALLSGLAHWVVANNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFANKAVALEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             complement(11808..11828)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(11872..11896)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(11893..11931)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    11943..11961
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11979..11997
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12014..12032
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12050..12068
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12086..12104
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12121..12139
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    complement(12184..12204)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    12249..12366
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(join(12664..12672,1..36))
                     /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"
ORIGIN
        1 ccatttatct aattccaata attcttgttc attcttttct tgctggtttt gcgattcttc
       61 aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact ccatccaggt
      121 cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc aaggcacagc
      181 agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt gatcctttag
      241 gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga ctgtgagttg
      301 caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct gcactatacc
      361 agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca tagtgcttcc
      421 tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt tttctctctg
      481 caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt ttactacttt
      541 atatttatat aattcacttc tccaattgtc cctcatatct cctcctccag gtctgaagat
      601 cagcggccgc ttgctgtgcg gtggtcttac ttttgttttg ctcttcctct atcttgtcta
      661 aagcttcctt ggtgtctttt atctctatcc tttgatgcac acaatagagg gttgctactg
      721 tattatataa tgatctaagt tcttctgatc ctgtctgaag ggatggttgt agctgtccca
      781 gtatttgtct acagccttct gatgtttcta acaggccagg attaactgcg aatcgttcta
      841 gctccctgct tgcccatact atatgtttta atttatattt tttctttccc cctggcctta
      901 accgaatttt ttcccatcgc gatctaattc tcccccgctt aatactgacg ctctcgcacc
      961 catctctctc cttctagcct ccgctagtca aaatttttgg cgtactcacc agtcgccgcc
     1021 cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag agctcctctg
     1081 gtttcccttt cgctttcaag tccctgttcg ggcgccactg ctagagattt tccacactga
     1141 ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac
     1201 ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga
     1261 gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa aacgcgctgc
     1321 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt
     1381 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa
     1441 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact
     1501 gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag
     1561 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga
     1621 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt
     1681 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa
     1741 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat
     1801 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa
     1861 ttgattacta ttaataacta gtcaataatc aatgtcaacg cgtatatctg gcccgtacat
     1921 cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg tcggtcaagc
     1981 cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc aacacacaag
     2041 cagggagcag atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     2101 ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt
     2161 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2221 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2281 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2341 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2401 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2461 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2521 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2581 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2641 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2701 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2761 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2821 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2881 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2941 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3001 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3061 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3121 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3181 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3241 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3301 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3361 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3421 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3481 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3541 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3601 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3661 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3721 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3781 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3841 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3901 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3961 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4021 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     4081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     4141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     4201 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     4261 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga
     4321 cggtatacag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4381 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4441 agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     4501 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     4561 ggtggaatcg aaatctcgta gcacgtgtca gtcctgctcc tcggccacga agtgcacgca
     4621 gttgccggcc gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc cgatctcggt
     4681 catggccggc ccggaggcgt cccggaagtt cgtggacacg acctccgacc actcggcgta
     4741 cagctcgtcc aggccgcgca cccacaccca ggccagggtg ttgtccggca ccacctggtc
     4801 ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc acaccggcga agtcgtcctc
     4861 cacgaagtcc cgggagaacc cgagccggtc ggtccagaac tcgaccgctc cggcgacgtc
     4921 gcgcgcggtg agcaccggaa cggcactggt caacttggcc atggtttagt tcctcacctt
     4981 gtcgtattat actatgccga tatactatgc cgatgattaa ttgtcaacac gtgctgatca
     5041 gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa
     5101 aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa
     5161 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     5221 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     5281 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     5341 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     5401 cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5461 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5521 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5581 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5641 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5701 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5761 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5821 cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag ctggttcttt
     5881 ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg tcttcccaat
     5941 cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac ctactcagac
     6001 aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca ccttccaggg
     6061 tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa ggcacagtcg
     6121 aggctgatca gcgggtttaa acgggccctg ctagagattt tccacactga ctaaaagggt
     6181 ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact
     6241 caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag
     6301 gctcagatct ggtctaacca gagagaccca gtacagtccg gatgcagctc tcgggccatg
     6361 tgatgaaatg ctaggcggct gtcaaacctc cactctaata cttctctctc cgggtcatcc
     6421 atcccatgca ggctcacagg gtgtaacaag cgggtgttct ctccttcatt ggcttcttct
     6481 accttctctt gctcaactgg tactagcttg tagcaccatc caaaggtcag tggatatctg
     6541 atccctggcc ctggtgtgta gttctgccaa tcagggaagt agccttgtgt gtggtagatc
     6601 cacagatcaa ggatatcttg tcttcgttgg gagtgaatta gcccttccag tccccccttt
     6661 tcttttaaaa agtggctaag atctacagct gccttgtaag tcattggtct taaaggtacc
     6721 ttagccctcc cacacataac cagagggcag caattcacga atcccaactg ccgtcggctg
     6781 tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa gcacctgtcg
     6841 gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga cgatacaagt
     6901 caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg cacaaggtcc
     6961 cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag agagagctgc
     7021 gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg ttgctctttc
     7081 aatgagggtg gattcttctt gagacaaagg cttggccatc ggtccaggat tctcttcgac
     7141 atctccggct tgtttcagca gagagaagtt tgttgcgccg gatcccccgg ggagcatgtc
     7201 aaggtcaaaa tcgtcaaggg catcggctgg gagcatgtct aagtcaaaat cgtcaagggc
     7261 gtcggtcggc ccgccgcttt cgcactttag ctgtttctcc aggccacata tgattagttc
     7321 caggccgaaa aggaaggcag gttcggctcc ctgccggtcg aacagctcaa ttgcttgtct
     7381 cagaagtggg ggcatagaat cggtggtagg tgtctctctt tcctcttttg ctacttgatg
     7441 ctcctgttcc tccaatacgc agcccagtgt aaagtggccc acggcggaca gagcgtacag
     7501 tgcgttctcc agggagaagc cttgctgaca caggaacgcg agctgatttt ccagggtttc
     7561 gtactgtttc tctgttgggc gggtgccgag atgcacttta gccccgtcgc gatgtgagag
     7621 gagagcacag cggtatgact tggcgttgtt ccgcagaaag tcttgccatg actcgccttc
     7681 cagggggcag aagtgggtat gatgcctgtc cagcatctcg attggcaggg catcgagcag
     7741 ggcccgcttg ttcttcacgt gccagtacag ggtaggctgc tcaactccca gcttttgagc
     7801 gagtttcctt gtcgtcaggc cttcgatacc gacaccattg agtaattcca gagctccgtt
     7861 tatgactttg ctcttgtcca gcctagacat ggtggcagcg ctctagaacc ggtcctgtgt
     7921 tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg
     7981 ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac
     8041 cccgcgccac cttctctagg caccggatca attgccgacc cctcccccca acttctcggg
     8101 gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagccaattc ccactccttt
     8161 caagacctac tcgaggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
     8221 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
     8281 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
     8341 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
     8401 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
     8461 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
     8521 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
     8581 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
     8641 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     8701 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     8761 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttgccggaat tcaccacttt
     8821 gtaccttaag cgtaatctgg aacatcgtat gggtaagcgg ccgccacctt cctctttttc
     8881 ttaggtccgg atccggaatt gccggacacc ttctttttct ccttgtcgaa cttggcggcc
     8941 tcgttcctat cgaacagggc ctcgatgctc aggttcttaa acctggggat acagtagccg
     9001 aaaggcacac acagcagttt caggagccta tcgttgtact tcttctcgtc attcacagcg
     9061 tcgaagtact cggacacctt tccgctgctt ttctcgtacc tctcattcat gataattctc
     9121 tgcatgatgt aatggtacag ttggaagtag gaattgacct cggcaatgtc gttgatatag
     9181 gcgtggacat acctggccac ttccagggcg acggccttgt ttgcgaacag ggtacatgtc
     9241 ttgttgtcaa ttctcaggag gtcctccctg atgatcacat tccacttggt gttcctcagg
     9301 taggcgttca gggcggtttt ggccttctcc ctgttaatct gcctggtgaa ctcctcggct
     9361 ttcttctcgt cgctgtattt gatgtaattg gcatacagct tggggttggc gctctcgagg
     9421 ctgtcaatgc tctccttggc tctttcagcc atctcctttt ccacgtcctt tctcttatcg
     9481 ggggcagttt catcaatgcc agcgcagagc ttggtgacgg agctgaatcc cttctcttcc
     9541 agtttcttga gattgatgtc gtagcctttc tccttgtaca gttgagcatc acgctcgacg
     9601 caatggaatc cgatgacgta cctggcgttg atattgacaa tattcttgag gatgtggtag
     9661 atcacggtga ggtacaggct gatgatcttt ttaaacttct ccctctcggc gttttccctg
     9721 ccggtgtcct caatgacgct ccttttcttg tcgaattggt cgtagttcat tccggtgatg
     9781 atctttgtga gagcgtccac cttttcgctc acgctcttgc ccttatcctt tccgatacaa
     9841 gtttcgtagt acctgtcgat ctggttcttg ccgttctggc cctgtttttt ctggatgtca
     9901 gcgatcctgc cgagcacgaa cttcaccacg gcctcgtttt tggcgatctc atggaggtgg
     9961 gcaggatcac cgtatctgat caggtagtgg aaccttttat tgctgatcac gttattaata
    10021 atgaaatttc tcatgccgtg cttgcctttc ttgagcttgt ttccgttctc gtccagggaa
    10081 aaggtgtcgg cgagggcctt gagctcatca taggacaggt tggttcctaa aatacggatg
    10141 gcgtcgatat acatggccct cctggcatcg gcaataggtt ctcccattct agcgaaggac
    10201 ttgatcagcc tcagctcatc ggcgatcttg gcggagtctt tgaaaaaggc gtattcctcc
    10261 acgaacttag cgttgactcc gatgagaggc atcaccttca ggaagctctg gatgttatcg
    10321 aatttattaa tcagggtggt caggaggtcg ttgatctcct tgccatccag gaacatggtc
    10381 agggcataca tgagtttgct gaaggcggaa acatcacggc cagcgggcag gattctgggc
    10441 agtctagggg cgtccttctt cttatactct cttgtcttgt ttcccctaaa ttccttgatg
    10501 ttgtgcatga tattttcgag ctttctccaa attctattag cttcatcgta gtagagggca
    10561 tccttctggt cgtcgttgaa ggagcccctc aggttaatca caaagatatc cttctcgctc
    10621 agggacttct cattatcggg gagggactta ttggcggcag ccaccttggc atcctcttcg
    10681 atgtaatacc tataaatcac aaagtccatc atggtgtaga ccttggtcct gatggagtcg
    10741 aacaccttat gatttttcct gatctcggac atatccttcc tgtccagcat cacttccctg
    10801 agcttggtga tattgaatcc gaggtttttc tgctctttca taatgctgaa tctgaaatat
    10861 tgttcggcga attcggcagg gttgattccc agagtttcgg caatatagtt cacgttggcg
    10921 gcggagttct tggagaagga gttggtcagc tcattggtga tcctgtcgta gaggtagttg
    10981 agggtggaga tgtattcgtt gtcgaggttc ttatcgaggt tgtagagcca ggtcctggag
    11041 atcctggact cttcttcgtt gttagcgacc acccagtgcg ccagtccgct caggagggcc
    11101 agaatgtcat agcattcgtt gccgtaattg atgatgtaat ttctgccctc cttgctgaaa
    11161 aaggcctggc cgaaatagcc gagtctgggg ttatcgagga agttgtcgaa ctcgtcatac
    11221 tgggccttga tggcgttgat gagcttatcg ttattgttga aagcggccct atggtgctcg
    11281 gggtctttga attcgtcgta ggtatacact gtggagaact tgccgaatcc aataatgtcc
    11341 ttatccaggc cggagatgtt gttcaccacg tacaccacgt ttgtgatgta ctccaccagg
    11401 atcttctcaa tgtccaggat gttatggatc acctggatac aaatattgtc attgccatca
    11461 gcgctctcgc cgaagtacct cttttccaga gtttccttca ggccgagcat atcctgctgg
    11521 acgggtcctg tatacagagg gttgttagcc accacggcgt agcccttagg atggctgaat
    11581 ttggcgttgc cgatcttata gccggcgttt ttatcggcca tttcagcgct gaaggcctcg
    11641 ccctcattca cgctcctgat gctgtcgccc tccacgatct tttccagcct ggcgtcgctg
    11701 ccttcggcga aggttgtcat gtacactttg gagccggaca cgagtgtgga cttcacgccc
    11761 atgcccttgg cgaaggactt ttttttttcg atgaattcgc tggcctccac ctttctcttc
    11821 ttcttggggc tcatggtggc tgtacaaact tgggtcgaat tcgccctttg cgatctgacg
    11881 gttcactaaa cgagctctgc ttatataggc ctcccaccgt acacgcctac ctcgacatac
    11941 gttctctatc actgataggg agtaaactcg acatacgttc tctatcactg atagggataa
    12001 actcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact
    12061 gatagggagt aaactcgaca tacgttctct atcactgata gggagtaaac tcgacatcgt
    12121 tctctatcac tgatagggag taaactcgtc tagaacctcg gacctgaagg gcgaattcgg
    12181 agccaacttt tctatacaaa gttgttaatt aaccaaactg gatctctgct gtccctgtaa
    12241 taaacccgaa aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct
    12301 gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt
    12361 ttaaaattgt ggatgaatac tgccatttgt ctgcagaatt ggcgcacgca gtgccgatcc
    12421 gttcactaat cgaatggatc tgtctctgtc tctctctcca ccttcttctt ctattccttc
    12481 gggcctgtcg ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc
    12541 ctgcctaact ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc
    12601 tcctactatc attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt
    12661 ccacaaactt gc
//
