LOCUS       sequence-259139-       13282 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral expression of CasRx T2A Hygromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13282)
  TITLE     Unpublished
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13282)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAK_EF1a_CasRx_Hygro
FEATURES             Location/Qualifiers
     source          1..13282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    61..178
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        303..1481
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          534..1472
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1429..1449
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1500..1520
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1530..4427
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             4437..4457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4467..4493
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4509..4562
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4563..5585
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4563..5585
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     misc_feature    5595..6183
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5648..5668)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6066..6077)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6186..6202)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6187..6203)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6708..6888
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6914..6931)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6920..7144
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7190..7618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7277..7296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7487..7508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7627..7647)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7632..7961
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7812..7947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7874..7893
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8009..8056
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8075..8449
                     /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    8579..8700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8616..8635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8670..8689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8749..8765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8749..8765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8762..8784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8773..8789
                     /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(8797..8803,8804..8821,8822..8827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8842..8863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8980..8997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9151..9739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9231..9250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9910..10701,10702..10770))
                     /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     10533..10552
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10771..10875)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10950..10969)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11141..11520
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11522..11720
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11675..11695
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11738..11918
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11965..12090
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12583..12816
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13001..13045
                     /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 gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa
       61 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
      121 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
      181 ggtttattac agggacagca gagatccagt ttggttaatt aatgcaaaga tggataaagt
      241 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      301 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      361 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      421 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      481 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      541 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      601 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      661 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      721 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      781 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      841 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      901 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      961 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     1021 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     1081 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     1141 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1201 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1261 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1321 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1381 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1441 cagtggttca aagttttttt cttccatttc aggtgtcgtg acgtacggcc accatgagcc
     1501 ccaagaagaa gagaaaggtg gaggccagca tcgaaaaaaa aaagtccttc gccaagggca
     1561 tgggcgtgaa gtccacactc gtgtccggct ccaaagtgta catgacaacc ttcgccgaag
     1621 gcagcgacgc caggctggaa aagatcgtgg agggcgacag catcaggagc gtgaatgagg
     1681 gcgaggcctt cagcgctgaa atggccgata aaaacgccgg ctataagatc ggcaacgcca
     1741 aattcagcca tcctaagggc tacgccgtgg tggctaacaa ccctctgtat acaggacccg
     1801 tccagcagga tatgctcggc ctgaaggaaa ctctggaaaa gaggtacttc ggcgagagcg
     1861 ctgatggcaa tgacaatatt tgtatccagg tgatccataa catcctggac attgaaaaaa
     1921 tcctcgccga atacattacc aacgccgcct acgccgtcaa caatatctcc ggcctggata
     1981 aggacattat tggattcggc aagttctcca cagtgtatac ctacgacgaa ttcaaagacc
     2041 ccgagcacca tagggccgct ttcaacaata acgataagct catcaacgcc atcaaggccc
     2101 agtatgacga gttcgacaac ttcctcgata accccagact cggctatttc ggccaggcct
     2161 ttttcagcaa ggagggcaga aattacatca tcaattacgg caacgaatgc tatgacattc
     2221 tggccctcct gagcggactg aggcactggg tggtccataa caacgaagaa gagtccagga
     2281 tctccaggac ctggctctac aacctcgata agaacctcga caacgaatac atctccaccc
     2341 tcaactacct ctacgacagg atcaccaatg agctgaccaa ctccttctcc aagaactccg
     2401 ccgccaacgt gaactatatt gccgaaactc tgggaatcaa ccctgccgaa ttcgccgaac
     2461 aatatttcag attcagcatt atgaaagagc agaaaaacct cggattcaat atcaccaagc
     2521 tcagggaagt gatgctggac aggaaggata tgtccgagat caggaaaaat cataaggtgt
     2581 tcgactccat caggaccaag gtctacacca tgatggactt tgtgatttat aggtattaca
     2641 tcgaagagga tgccaaggtg gctgccgcca ataagtccct ccccgataat gagaagtccc
     2701 tgagcgagaa ggatatcttt gtgattaacc tgaggggctc cttcaacgac gaccagaagg
     2761 atgccctcta ctacgatgaa gctaatagaa tttggagaaa gctcgaaaat atcatgcaca
     2821 acatcaagga atttagggga aacaagacaa gagagtataa gaagaaggac gcccctagac
     2881 tgcccagaat cctgcccgct ggccgtgatg tttccgcctt cagcaaactc atgtatgccc
     2941 tgaccatgtt cctggatggc aaggagatca acgacctcct gaccaccctg attaataaat
     3001 tcgataacat ccagagcttc ctgaaggtga tgcctctcat cggagtcaac gctaagttcg
     3061 tggaggaata cgcctttttc aaagactccg ccaagatcgc cgatgagctg aggctgatca
     3121 agtccttcgc tagaatggga gaacctattg ccgatgccag gagggccatg tatatcgacg
     3181 ccatccgtat tttaggaacc aacctgtcct atgatgagct caaggccctc gccgacacct
     3241 tttccctgga cgagaacgga aacaagctca agaaaggcaa gcacggcatg agaaatttca
     3301 ttattaataa cgtgatcagc aataaaaggt tccactacct gatcagatac ggtgatcctg
     3361 cccacctcca tgagatcgcc aaaaacgagg ccgtggtgaa gttcgtgctc ggcaggatcg
     3421 ctgacatcca gaaaaaacag ggccagaacg gcaagaacca gatcgacagg tactacgaaa
     3481 cttgtatcgg aaaggataag ggcaagagcg tgagcgaaaa ggtggacgct ctcacaaaga
     3541 tcatcaccgg aatgaactac gaccaattcg acaagaaaag gagcgtcatt gaggacaccg
     3601 gcagggaaaa cgccgagagg gagaagttta aaaagatcat cagcctgtac ctcaccgtga
     3661 tctaccacat cctcaagaat attgtcaata tcaacgccag gtacgtcatc ggattccatt
     3721 gcgtcgagcg tgatgctcaa ctgtacaagg agaaaggcta cgacatcaat ctcaagaaac
     3781 tggaagagaa gggattcagc tccgtcacca agctctgcgc tggcattgat gaaactgccc
     3841 ccgataagag aaaggacgtg gaaaaggaga tggctgaaag agccaaggag agcattgaca
     3901 gcctcgagag cgccaacccc aagctgtatg ccaattacat caaatacagc gacgagaaga
     3961 aagccgagga gttcaccagg cagattaaca gggagaaggc caaaaccgcc ctgaacgcct
     4021 acctgaggaa caccaagtgg aatgtgatca tcagggagga cctcctgaga attgacaaca
     4081 agacatgtac cctgttcaga aacaaggccg tccacctgga agtggccagg tatgtccacg
     4141 cctatatcaa cgacattgcc gaggtcaatt cctacttcca actgtaccat tacatcatgc
     4201 agagaattat catgaatgag aggtacgaga aaagcagcgg aaaggtgtcc gagtacttcg
     4261 acgctgtgaa tgacgagaag aagtacaacg ataggctcct gaaactgctg tgtgtgcctt
     4321 tcggctactg tatccccagg tttaagaacc tgagcatcga ggccctgttc gataggaacg
     4381 aggccgccaa gttcgacaag gagaaaaaga aggtgtccgg caattccgga tccggaccta
     4441 agaaaaagag gaaggtggcg gccgcttacc catacgatgt tccagattac gctgctagcg
     4501 gcagtggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
     4561 caatgaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     4621 gcgtgtccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     4681 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     4741 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     4801 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     4861 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     4921 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     4981 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     5041 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     5101 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     5161 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     5221 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     5281 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     5341 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     5401 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     5461 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     5521 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     5581 aataagtcta gaataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5641 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     5701 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     5761 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     5821 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     5881 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     5941 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     6001 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     6061 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     6121 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     6181 cgcatcgata ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata
     6241 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     6301 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     6361 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     6421 atatccttga tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca
     6481 caccagggcc agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     6541 ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg
     6601 tgagcctgca tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc
     6661 gcctagcatt tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt
     6721 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6781 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6841 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa
     6901 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     6961 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     7021 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     7081 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     7141 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     7201 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     7261 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     7321 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     7381 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     7441 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     7501 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     7561 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     7621 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     7681 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     7741 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     7801 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     7861 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     7921 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     7981 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     8041 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
     8101 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     8161 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     8221 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     8281 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     8341 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     8401 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
     8461 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     8521 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     8581 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8641 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     8701 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     8761 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     8821 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     8881 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     8941 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     9001 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     9061 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     9121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     9181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     9241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     9301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     9361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     9421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     9481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     9541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     9601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     9661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     9721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     9781 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     9841 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     9901 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     9961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
    10021 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
    10081 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
    10141 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
    10201 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
    10261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
    10321 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
    10381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    10441 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
    10501 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    10561 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    10621 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    10681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    10741 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    10801 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    10861 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
    10921 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
    10981 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
    11041 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
    11101 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
    11161 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
    11221 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
    11281 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
    11341 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
    11401 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
    11461 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
    11521 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
    11581 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
    11641 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
    11701 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
    11761 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
    11821 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
    11881 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
    11941 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
    12001 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
    12061 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
    12121 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
    12181 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
    12241 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
    12301 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
    12361 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
    12421 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
    12481 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
    12541 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
    12601 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
    12661 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
    12721 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
    12781 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
    12841 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
    12901 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
    12961 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
    13021 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
    13081 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
    13141 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
    13201 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
    13261 cagagacaga tccattcgat ta
//
