LOCUS       sequence-375469-       14335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of NLS-NLS-dMisCas13b-3xGFP-NLS in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14335)
  AUTHORS   Yang LZ, Gao BQ, Huang Y, Wang Y, Yang L, Chen LL
  TITLE     Multi-color RNA imaging with CRISPR-Cas13b systems in living cells.
  JOURNAL   Cell Insight. 2022 Jun 15;1(4):100044. doi: 
            10.1016/j.cellin.2022.100044. eCollection 2022 Aug.
  PUBMED    37192858
REFERENCE   2  (bases 1 to 14335)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Insight."; date: "2022-06-15"; volume: "1(4):100044. doi"; pages: " 
            10.1016/j.cellin.2022.100044. eCollection 2022 Aug"
COMMENT     Sequence Label: pHAGE-IRES-puro-NLS-NLS-dMisCas13b-3xGFP-NLS
FEATURES             Location/Qualifiers
     source          1..14335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2223..3401
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3392
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3349..3369
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3414..3461
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3471..3491
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7293..8006
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             8025..8738
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             8757..9470
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             9477..9497
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    9514..10087
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9681..9698)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9908..9927
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             10107..10706
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(10107..10126)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10603..10623
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10725..11313
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10778..10798)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11196..11207)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11388..11621
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11740..11758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11826..11930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11931..11999,12000..12791)
                     /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(12149..12168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12962..13550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13451..13470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13704..13721
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13961..13982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13997..14002,14003..14020,14021..14027)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14035..14051
                     /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     14040..14062
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     14059..14075
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14059..14075
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     14125..14144
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccgc atgaagcgac
     3421 ctgccgccac aaagaaggct ggacaggcta agaagaagaa agctagcatg cccaaaaaga
     3481 agaggaaagt gttcgtgaga tatagccctg agtcctggcc taggcacttc cacggcctgc
     3541 agcagagact gcccaccccc atcggccagc acaccttcct gagtgataac tttaacacca
     3601 tggacatccg gctgagtaag aatagagccc tgctggcagc ttacctgaat atggcccagc
     3661 agaacgtgta cgtgaccctg ctgcacatct cccagctgat gggagtggag gagaccaaag
     3721 acctgtctaa ccgcatgtac gactttcaga ttctgcagga gcttaagacc gggaagaagg
     3781 gcgccgagtc tgagcagatg gtgaagtctg agagagtgca gaagatcatt ctcctgctgt
     3841 atcgcaactt ccctttcctg aaggccttcg tggaggctaa gtacaagcag caggagaaga
     3901 acaatggggg caaaaacaag ggcggcggaa aaaacacaca gaagaataag aaaacccctg
     3961 acaatacaag caacgaggag cagctggagt tcactgagag tgagctgggc aagctgccca
     4021 ttccaccaca tgagtacaga accatcctgt gtgatgtgtt caggctgatc aatgacctgg
     4081 ccaatgagta cacagccttc agccctgaga aacgcactgg cggcgctaag gagtcccacg
     4141 aaaagtgcca gaagaagatg ctgagctacc tgtacctgtg cctggacgga ggcagaaggg
     4201 tggtgaagga gcggttcaag gacaccttcc aagagaagga ctttaacttt ctcacccagg
     4261 gcccaagata cagcctccag aaagacggag ataagaagaa ggccgaggaa agacctaatt
     4321 tttactacag actgaaggat gaccagggac tgttctctta cgtcggtatc ctcctgttca
     4381 tcagcctgtt cctgcacaag aggtatattt ccgagatgct ggcccagtcc gggctgtttt
     4441 atacacctcg gtactggaag ggccagccag tgaacgtgga actggagagg aggatcatca
     4501 gtgagatctt ttccatctat cgcatcaggc tgcccaagga gagaatcgac tccaccagac
     4561 cagattacgc cctcggcctg gatatgctga atgagctgca gaagtgccct aatgagctgt
     4621 atgagacctt ctcacctaaa gaccaggctg agtttagagt gaagcgggag tatgacgaga
     4681 gcggcgaagg cacagaagag ggcgaggtgc tgctgaaaag attcgaggac agattccctt
     4741 atttcgccat tcgctatctg gacgagaata gagtgttcga cgacatcaga ttccaggtga
     4801 gcatcggcaa gtataggtac aaattctaca ataagctgtg catcgacgtg gtgagcagca
     4861 atcggaacga tgactgctat cgcctgaggt ccctgcagaa agagattaat ggcttcggca
     4921 gactgaatga agtggagtac gagagaatcc tgagatacgg gtctcacatc cacgtggaga
     4981 agcctgaggg cgacgacacc ccctacctga ctgaccatca taccagctac ctgtttaacg
     5041 gcaataggat cggcctgctg ttcaaccagg agaacggcag gcacgatctg gaggagatca
     5101 gaggaaactg cctgtatctg cctcacctga cttttgcatc ctcctccgag ggaaaggatg
     5161 ccaggcgcgt gccccccatc gcctggctgt ccacctatga gctgcctgcc ctgatcttcc
     5221 accacctgct gtgcagacac tatggcctgg agagaactgc caccgagagc atcatcaagg
     5281 aatgtgtggg cctgtaccac cggctgttta ctgatatccg cgacggtaaa ctgctccccg
     5341 gagaggccct gggcatccag gccgaggagt ccacaaggag cgagattagt aggatcaaga
     5401 agggtgaacc cctgccccag aacatctctg aaaagattga aaccttcctg agcactcact
     5461 actgcaccga acaggtgtgc ctgcacatcc aggacattcc taagaaaatc atcgattacc
     5521 tgatcgggaa aaacatcgac caggacatga gcggactctt taagcagaga agcagagagc
     5581 ggattaataa gatgtatgag tccactaaaa ttcgcctcaa gaagttcaag accgacctgg
     5641 acctgcaggg caacggcgac aataagtttg gaaagaagag gtacactgac attaggccag
     5701 gacagctggc cgactatctg gccaaggacc tgctgttctt ccagccatat gacgacgaga
     5761 agaagaataa ggtcactgga ctgaactatc agatcctgca ggccgcactg gccatctaca
     5821 atcacaacct ggatgagctc agacgactct tcaaaaatgc ccggctgctc gtgcccgaga
     5881 atcctaagat ggagcaccct ttcctgggct ctgtgctgca caagtccccc cggaccactc
     5941 tgcagttcta cgcctactac ctgctcgaga agatctctta cctgaaaaag agactcgacg
     6001 atcctaacga gacattcgcc gatctgttta gatataagca gaggcagaaa tggcagacta
     6061 gagacagcca gtggtatagg gccttggctg gcaggtacat ggctgagatg atggaagagg
     6121 gcaccatgaa ttttaaaccc gtggagctgc caagagggct gtttgaaaga cctatcaagc
     6181 ggctgctgac caatatcctg actaagcgga ccgacagcga tagccaggcc ctgctgaaga
     6241 tcgtggagag ccccaggtgc aacgtgagct acctgattgc tcagtatttc actctgattc
     6301 tgcacgacga ttcccagccc ttttaccacc ctaacaaacg ctacaaaagg acctaccagt
     6361 atttcaacat cctgaacaat agaaaggagc ggaataagct cgtgcaggtg tactacactc
     6421 tggatgagat ggaggcttac tttaaggata ccgaatctct gaagaagagg tataaggact
     6481 acctgactag ctccgaggct aagaagcacc cagcccccga gcagaggctg aagaagtatc
     6541 tgtcagagta tcgcaagaat gagaccacca ttagacggta cagagtgcag gacatgctgg
     6601 tgtttctgat ggccaaaatt ctgctgctgg aaaagggcgg cctcgaagcc ggcgagctgg
     6661 agcagtacaa gctgtcccac attcagccca ccaccggcca gaacagcggc cagaacatcc
     6721 tgagcctgca ggtgcccttt aacatcaccg tgaagctgga cgatggcaca gagagaagca
     6781 tcacccagga agtgatcaag gtgaaaaatt atggcgactt ctttagattt ctctatgatg
     6841 accgggtggc atccctgctg ccttatactc aggaggacat tcccctggag cgggccctgc
     6901 tggagaagga gctggaggga tacgacaggg agagaatccc tattttcaag agcgtgcact
     6961 gcatggagaa ggagatcctg aggctgtgcc ctgaactgaa tgatccccag tccagccagt
     7021 acaagtatac cacagatatt ggcactgaga agatcgtgaa ggagaatttc agggccatgc
     7081 tgactgagct gtgtagccgg ggcctgctcc ctgagaacga gatcgagatg atggtggaga
     7141 tcgccaatag cttttccgcc aatagatacg cctgcgtgaa cctgaagggc aagaatatcc
     7201 caatgatcgc catctccatc cagcggaagt tcgacggcgt ggtgggcaga accatcggaa
     7261 agaataagaa cgcgggaggt ggatccggaa gtatgcgtaa aggcgaagag ctgttcactg
     7321 gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag ttttccgtgc
     7381 gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc atctgtacta
     7441 ctggtaaact gccggtacct tggccgactc tggtaacgac gctgacttat ggtgttcagt
     7501 gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc gccatgccgg
     7561 aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac aaaacgcgtg
     7621 cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa ggcattgact
     7681 ttaaagaaga cggcaatatc ctgggccata agctggaata caattttaac agccacaatg
     7741 tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa attcgccaca
     7801 acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact ccaatcggtg
     7861 atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt ctgtctaaag
     7921 atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca gcgggcatca
     7981 cgcatggtat ggatgaactg tacaaatctg gcggccgctc tggcatgcgt aaaggcgaag
     8041 agctgttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata
     8101 agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt
     8161 tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg acgctgactt
     8221 atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt
     8281 ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt
     8341 acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga
     8401 aaggcattga ctttaaagaa gacggcaata tcctgggcca taagctggaa tacaatttta
     8461 acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta
     8521 aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca
     8581 ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg
     8641 ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg
     8701 cagcgggcat cacgcatggt atggatgaac tgtacaaagg tagcatcgat ggtagcatgc
     8761 gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg
     8821 tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac
     8881 tgacgctgaa gttcatctgt actactggta aactgccggt accttggccg actctggtaa
     8941 cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg
     9001 acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg
     9061 atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc
     9121 gcattgagct gaaaggcatt gactttaaag aagacggcaa tatcctgggc cataagctgg
     9181 aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta
     9241 aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact
     9301 accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga
     9361 gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg
     9421 agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa ctcgagccaa
     9481 agaaaaagcg gaaagtgtaa tctagaggat ctgcccctct ccctcccccc cccctaacgt
     9541 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     9601 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     9661 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     9721 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     9781 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     9841 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     9901 agtcaaatgg ctctcctcaa gcgtattcaa caatgggctg aaggatgccc agaaggtacc
     9961 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
    10021 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
    10081 atgataagga tccaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca
    10141 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
    10201 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
    10261 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
    10321 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
    10381 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
    10441 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
    10501 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
    10561 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
    10621 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
    10681 gcatgacccg caagcccggt gcctgacgcc catcgataga tcctaatcaa cctctggatt
    10741 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
    10801 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
    10861 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
    10921 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
    10981 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
    11041 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
    11101 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
    11161 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
    11221 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
    11281 cgagtcggat ctccctttgg gccgcctccc cgcctgagat cctttaagac caatgactta
    11341 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
    11401 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
    11461 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
    11521 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
    11581 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
    11641 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt
    11701 aattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata cgcctatttt
    11761 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
    11821 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
    11881 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
    11941 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
    12001 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
    12061 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
    12121 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
    12181 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
    12241 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
    12301 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
    12361 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
    12421 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
    12481 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
    12541 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
    12601 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
    12661 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
    12721 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
    12781 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
    12841 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
    12901 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    12961 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    13021 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    13081 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
    13141 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    13201 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    13261 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    13321 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    13381 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    13441 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    13501 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    13561 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    13621 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    13681 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
    13741 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag ctcatggctg
    13801 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
    13861 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg tggcacgaca
    13921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
    13981 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
    14041 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc acaaataaag
    14101 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
    14161 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc
    14221 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat
    14281 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
