LOCUS       sequence-332232-       12201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  overexpression of RfxCas13d in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12201)
  AUTHORS   Li S, Li X, Xue W, Zhang L, Yang LZ, Cao SM, Lei YN, Liu CX, Guo SK,
            Shan L, Wu M, Tao X, Zhang JL, Gao X, Zhang J, Wei J, Li J, Yang L, 
            Chen LL
  TITLE     Screening for functional circular RNAs using the CRISPR-Cas13 
            system.
  JOURNAL   Nat Methods. 2021 Jan;18(1):51-59. doi: 10.1038/s41592-020-01011-4. 
            Epub 2020 Dec 7.
  PUBMED    33288960
REFERENCE   2  (bases 1 to 12201)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-020-01011-4"; journalName: "Nat Methods"; date: 
            "2021-01"; volume: "18"; issue: "1"; pages: "51-59"
COMMENT     Sequence Label: p23-NLS-RfxCas13d-mcherry-NLS-Flag
FEATURES             Location/Qualifiers
     source          1..12201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1192..1239
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             1246..4143
                     /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"
     primer_bind     4563..4582
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4861..4881
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4897..4962
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    4969..5536
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5130..5147)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5357..5376
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5556..6350
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5556..6350
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5610..5629)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6220..6239
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    6369..6957
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6422..6442)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6840..6851)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7032..7265
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7384..7402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7470..7574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7575..7643,7644..8435)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7793..7812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8606..9194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9095..9114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9348..9365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9605..9626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9641..9646,9647..9664,9665..9671)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9679..9695
                     /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     9684..9706
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9703..9719
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9703..9719
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9769..9788
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9980..10613
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10660..10785
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11282..11515
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11700..11744
                     /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    12037..12154
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa gcggccgcat gaagcgacct
     1201 gccgccacaa agaaggctgg acaggctaag aagaagaaag ctagcatcga gaagaagaag
     1261 agcttcgcca agggcatggg cgtgaagagc accctggtgt ccggctctaa ggtgtacatg
     1321 accacatttg ccgagggctc cgacgcacgg ctggagaaga tcgtggaggg cgatagcatc
     1381 agatccgtga acgagggaga ggccttcagc gccgagatgg ccgacaagaa tgccggctat
     1441 aagatcggca acgccaagtt ttcccaccca aagggctatg ccgtggtggc caacaatcca
     1501 ctgtacaccg gaccagtgca gcaggacatg ctgggcctga aggagacact ggagaagaga
     1561 tacttcggcg agagcgccga cggcaatgat aacatctgca tccaggtcat ccacaatatc
     1621 ctggatatcg agaagatcct ggccgagtat atcaccaacg ccgcctacgc cgtgaacaat
     1681 atctccggcc tggacaagga tatcatcggc ttcggcaagt tttctaccgt gtacacatat
     1741 gacgagttca aggaccccga gcaccacagg gccgccttta acaataacga caagctgatc
     1801 aatgccatca aggcccagta cgacgagttc gataattttc tggataaccc ccggctgggc
     1861 tattttggcc aggccttctt ttctaaggag ggcagaaact atatcatcaa ttacggcaac
     1921 gagtgttacg acatcctggc cctgctgagc ggactgaggc actgggtggt gcacaataac
     1981 gaggaggagt ctaggatcag ccgcacctgg ctgtataatc tggacaagaa tctggataac
     2041 gagtacatct ccacactgaa ctacctgtat gatcggatca ccaatgagct gacaaacagc
     2101 ttctccaaga attctgccgc caatgtgaac tacatcgcag agaccctggg catcaaccca
     2161 gcagagttcg cagagcagta cttccggttc agcatcatga aggagcagaa gaatctgggc
     2221 tttaacatca caaagctgcg ggaagtgatg ctggacagaa aggatatgtc cgagatccgg
     2281 aagaaccaca aggtgttcga ttctatcaga accaaggtgt atacaatgat ggactttgtg
     2341 atctaccggt actatatcga ggaggatgcc aaggtggccg ccgccaataa gagcctgccc
     2401 gacaacgaga agtctctgag cgagaaggat atcttcgtga tcaatctgag aggctccttt
     2461 aacgacgatc agaaggacgc cctgtactat gatgaggcca atcggatctg gagaaagctg
     2521 gagaatatca tgcacaacat caaggagttc aggggcaaca agacccgcga gtacaagaag
     2581 aaggacgcac caaggctgcc tcgcatcctg cctgcaggaa gggacgtgag cgccttcagc
     2641 aagctgatgt atgccctgac aatgtttctg gacggcaagg agatcaatga tctgctgacc
     2701 acactgatca ataagttcga caacatccag tcctttctga aagtgatgcc tctgatcggc
     2761 gtgaacgcca agttcgtgga ggagtacgcc ttctttaagg acagcgccaa gatcgccgat
     2821 gagctgaggc tgatcaagtc ctttgccagg atgggagagc caatcgcaga cgcaaggaga
     2881 gccatgtaca tcgatgccat cagaatcctg ggcaccaatc tgtcttatga tgagctgaag
     2941 gccctggccg acaccttcag cctggacgag aatggcaaca agctgaagaa gggcaagcac
     3001 ggcatgcgca atttcatcat caataacgtg atcagcaaca agaggtttca ctatctgatc
     3061 agatacggcg accctgcaca cctgcacgag atcgccaaga acgaggccgt ggtgaagttc
     3121 gtgctgggca ggatcgccga tatccagaag aagcagggcc agaatggcaa gaaccagatc
     3181 gaccgctact atgagacctg catcggcaag gataagggca agtccgtgtc tgagaaggtg
     3241 gacgccctga ccaagatcat cacaggcatg aattatgacc agttcgataa gaagcggagc
     3301 gtgatcgagg acaccggaag ggagaacgca gagagagaga agtttaagaa gatcatcagc
     3361 ctgtatctga cagtgatcta ccacatcctg aagaatatcg tgaatatcaa cgccagatac
     3421 gtgatcggct tccactgcgt ggagcgcgat gcccagctgt ataaggagaa gggctacgac
     3481 atcaacctga agaagctgga ggagaagggc tttagctccg tgaccaagct gtgcgcagga
     3541 atcgacgaga cagcacccga caagcggaag gatgtggaga aggagatggc cgagagagcc
     3601 aaggagagca tcgactccct ggagtctgcc aatcctaagc tgtacgccaa ctatatcaag
     3661 tactccgatg agaagaaggc cgaggagttc acccggcaga tcaatagaga gaaggccaag
     3721 accgccctga acgcctacct gcggaataca aagtggaacg tgatcatcag ggaggacctg
     3781 ctgcgcatcg ataataagac ctgtacactg ttcaggaaca aggccgtgca cctggaggtg
     3841 gcccgctatg tgcacgccta catcaatgac atcgccgagg tgaactccta ctttcagctg
     3901 taccactata tcatgcagcg gatcatcatg aacgagagat atgagaagtc tagcggcaag
     3961 gtgtctgagt acttcgacgc cgtgaatgat gagaagaagt ataacgatag actgctgaag
     4021 ctgctgtgcg tgcctttcgg ctactgtatc ccaaggttta agaatctgag catcgaggcc
     4081 ctgttcgaca ggaacgaggc agccaagttt gataaggaga agaagaaggt gagcggcaac
     4141 tccagtctag aggtgtccaa gggcgaagag gataacatgg ccatcatcaa ggagttcatg
     4201 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     4261 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     4321 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     4381 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     4441 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     4501 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     4561 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     4621 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     4681 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     4741 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     4801 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtac
     4861 cccaagaaaa agcgcaaggt acgcgtgcag ctaagcgact acaaagacca tgacggtgat
     4921 tataaagatc atgacatcga ttacaaggat gacgatgaca agtaaggatc cctccccccc
     4981 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     5041 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     5101 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     5161 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     5221 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     5281 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     5341 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     5401 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     5461 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     5521 aaaaacacga tgataaggat ccaccggagc ttaccatgat tgaacaagat ggattgcacg
     5581 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5641 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5701 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     5761 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5821 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5881 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5941 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     6001 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6061 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     6121 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6181 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6241 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6301 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga cgcccatcga
     6361 tagatcctaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     6421 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     6481 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     6541 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     6601 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     6661 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6721 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     6781 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     6841 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     6901 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     6961 agatccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     7021 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     7081 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7141 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7201 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7261 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     7321 atcagagagt gagaggcccg ggttaattaa ggaaagggct agatcattct tgaagacgaa
     7381 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     7441 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     7501 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     7561 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     7621 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     7681 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     7741 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7801 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7861 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     7921 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7981 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     8041 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     8101 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     8161 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     8221 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     8281 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     8341 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     8401 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     8461 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     8521 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     8581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     8641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     8701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     8761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     8821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     8881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     8941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     9001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     9061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     9121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     9181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     9241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     9301 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     9361 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     9421 attaatgcag caagctcatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     9481 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     9541 caaaaagctc cccgtggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     9601 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     9661 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca
     9721 tgattacgaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     9781 aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc
     9841 atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac
     9901 tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat
     9961 gtactacaaa cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt
    10021 gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg
    10081 ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca
    10141 gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg
    10201 catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca
    10261 tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct
    10321 tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg
    10381 gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct
    10441 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
    10501 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
    10561 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
    10621 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
    10681 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
    10741 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
    10801 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
    10861 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
    10921 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
    10981 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
    11041 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
    11101 agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata
    11161 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
    11221 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
    11281 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
    11341 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
    11401 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
    11461 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
    11521 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
    11581 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
    11641 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
    11701 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
    11761 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
    11821 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
    11881 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
    11941 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
    12001 atcgccgaat tcacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
    12061 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
    12121 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
    12181 tccagtttgg actagtcgtg a
//
