LOCUS       sequence-375471-       13925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of NLS-NLS-dHgm4Cas13b-3xGFP-NLS in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13925)
  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 13925)
  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-dHgm4Cas13b-3xGFP-NLS
FEATURES             Location/Qualifiers
     source          1..13925
                     /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             6882..7595
                     /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             7614..8327
                     /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             8346..9059
                     /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             9066..9086
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    9103..9676
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9270..9287)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9497..9516
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             9696..10295
                     /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(9696..9715)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10192..10212
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10314..10902
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10367..10387)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10785..10796)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10977..11210
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11329..11347)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11415..11519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11520..11588,11589..12380)
                     /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(11738..11757)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12551..13139
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13040..13059
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13293..13310
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13550..13571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13586..13591,13592..13609,13610..13616)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13624..13640
                     /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     13629..13651
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13648..13664
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13648..13664
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13715..13734
                     /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 gatgaagatc ccccagatca tcgagaacaa caagcacctg ttcggcgcct
     3541 acagcactat ggccctgagc aacattcgca acatcctgga tcacatcgcc accctggctg
     3601 acatcgaaaa tgacttctac agcgatagcg actcttattg gtaccaccca tgcatcaaga
     3661 tcatcgaccc tcggaacaac tgcaccgacg ccgccaaggc tgatttcgtg atcgagaaga
     3721 ttaagtctca tttcccattc gtgccaatcg tggccgaggc ccagaggcag tccgatatcc
     3781 tgagagccaa gaagcagctg gacatcgccg tgagaaatag ggattaccag aagcaggagg
     3841 agctgaagca ccggcagaag tccctgctgt ccgtgaccaa tgccgatatc taccgggtgc
     3901 tgaataacat gctgtgggtg ctggccaact acgccaacca gaccgtggcc tacctgatca
     3961 gcgacatcaa gttcgaagat ggttccgact ttctcaatta taacgagcag cctctgtccc
     4021 ggatgctgaa taactacttc accatcgccc tccgggacac cgctaacaag tacagctata
     4081 gccctgaggc cctgagcttc atccagtcaa gtaggtataa gactgagaat cggcggaaaa
     4141 tcctggacac agactttttc ctgtctatcc agcacaacaa cagcgatact aaccctaaga
     4201 accttcacct gtccggcgtg ggcgtggccc tgctgatttg tctgtttctg gagaaaaaat
     4261 atgtgaatgt gtttctgcag aagctgccca tctacggacc ctacaagaag cagtccatgg
     4321 aggccaacat catcagacag agcttttcca tccacaccgt gaagctgcca aaggagcgga
     4381 ttgtctctga aaagagcgac ttcaccattg cactggatat gctgaacgag ctgaagagat
     4441 gccccaagca gttgtttagc accctgtcct acgctgacca gaacgccttc agaatcgtga
     4501 gcagtgatat gagcgacgtg ctgcaggtga ggcatactga ccggttcgcc cagttaagcc
     4561 tggagtacat cgatcggcgg gaactgttta gcgacattag atttcacctg aacatgggca
     4621 agctgcggta cctgaaaacc gctgacaagc actgcatcga tggtgtgtct agggtgagag
     4681 tgctggaggg taaaattaac gcctttggcc gcattcatga gtttgaggcc aggcggaagg
     4741 agcagggatt cgtggagtgg cacgagcagg cagcccggct gatcaagacc aataagaata
     4801 tcgaaatcag ggactttgag cacgtgaagc gcgacgacgc cacacccgat agctacccct
     4861 acattattga cacttacacc cactatctgc tggagaataa taagatcgag atgaggtttg
     4921 gcgactacct cccaaaactg gtgaaggtgg acgaccagaa gtggaccgtg tacaacgaaa
     4981 tgcccacatg ttgcatgtcc gccctggagc tgcccgccat gatgttccac atgatgctgt
     5041 gcggcagcga agcaacagag catctgatca aggccgaggt ggagaagtat aagaagctct
     5101 ttggcgccat ggctaacggc acactgtcta aggagaatat cggcaacttc gagattgcca
     5161 tggaggacct gccaaagaaa gtgatcgact gcgtgaatag gaagaagtct gggaagtctt
     5221 ttgtcaagca gattaagaag gagatcgccg agatgctggc cgacaccaac ctgaggattg
     5281 tgagactgaa gtccgataag cgcagcgtgg cctccaccca gaataagatg ggcaaacggg
     5341 gctttagaag cattcagccc ggcaagctgg ccgactggct cgccgccgat atcgtgaaac
     5401 accagcctag cctgtttagc ggcgacgact acggtaccga taggattacc ggcatgaatt
     5461 accgggtcat gcagtctact atcgctacct tcaatgctgc cacccccgag cacagcctgg
     5521 acgagctgaa aagagtgttt agcgccgcac agctgattca atgtcaggag agggagcacc
     5581 cttttctcca gaaggccctg aatcgcaatc cccagaacac catcgacttc tacgagttct
     5641 atctgcaggc taagagagct tactatcagt ccctgcggca gtctatcgag aacggcgagg
     5701 acgtgaaact gccatacctg aaccctgatc gcaataagtg ggtgaggaga ggggccagct
     5761 tttacagcac catgggagat atctacctgg aggacatgcc cattgagctg ccaaggcaga
     5821 tgttcgacaa ggagatcaag aaggccctgg cccagctgcc ttccatgaag ggcgtggata
     5881 tgaagcagag caacgtgacc ttcctgatcg ccgagtacct caagagagaa ctgaaggatg
     5941 actttcagcc attttatcag tggaacagaa attacaggtt tacagacatg atgatttgcg
     6001 aggagaactg cgacaccaag gccctggcat ccaaattcat ccctctggag ctccgcgagg
     6061 agatctggga gaagcggggc gagctgaaag atgcctacaa aagatgggct ttcggcagac
     6121 tgaataagaa tcctgatatc aagcggctga gccctgccga gaaggacgaa ctgctggaaa
     6181 agcgcatcgc caagtgtcgc aacgagtacc agaagtccga gaagatcatt cggagataca
     6241 aggtgcagga tgtcctgatg ttcatgatgg tcaagtccat gtttgctcag ggagtgttca
     6301 ccgccgagtc caaggagttt gccctgtcca caatcacccc tgacgccgag cggggcattc
     6361 tgtccgaagt ggtgcccatc gatttcaagt tttccatcga cggaaagacc tacaccatcc
     6421 actccaacgg catgaagatc aagaactacg gcgattttta caagctgatc aacgacaaga
     6481 gaatgaagtc tctgctgaat atcgtgaccc actctactat cgagaaggac ctgctggaga
     6541 aggagtttag cacctacgac gataagagac ctgaggccgt ggagatcatc ttcgagtttg
     6601 agaaggccgc ctacgagaag caccccgagc tgaaggagct gctgactccc gagcgcaagt
     6661 ttgatttcag ggccctgctg gaggagctcc aggccaagtc tgacctgtct cagatcgatg
     6721 ccagatatct gagccagatc gccaacgcct tcaacgccaa cgcctacccc cggaatctga
     6781 agatcacatc caacatccct gagatcgccc aggagatggt gaacatcttc aggatcaccg
     6841 acccctacaa gaaaggcaag aagggaggtg gatccggaag tatgcgtaaa ggcgaagagc
     6901 tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac ggtcataagt
     6961 tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg ctgaagttca
     7021 tctgtactac tggtaaactg ccggtacctt ggccgactct ggtaacgacg ctgacttatg
     7081 gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc ttcaagtccg
     7141 ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac ggcacgtaca
     7201 aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt gagctgaaag
     7261 gcattgactt taaagaagac ggcaatatcc tgggccataa gctggaatac aattttaaca
     7321 gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg aattttaaaa
     7381 ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag caaaacactc
     7441 caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg caaagcgttc
     7501 tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc gtaaccgcag
     7561 cgggcatcac gcatggtatg gatgaactgt acaaatctgg cggccgctct ggcatgcgta
     7621 aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat ggtgatgtca
     7681 acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat ggtaaactga
     7741 cgctgaagtt catctgtact actggtaaac tgccggtacc ttggccgact ctggtaacga
     7801 cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag cagcatgact
     7861 tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc tttaaggatg
     7921 acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg gtaaaccgca
     7981 ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat aagctggaat
     8041 acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat ggcattaaag
     8101 cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct gatcactacc
     8161 agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac tatctgagca
     8221 cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt ctgctggagt
     8281 tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaaggt agcatcgatg
     8341 gtagcatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg
     8401 atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta
     8461 atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta ccttggccga
     8521 ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga
     8581 agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt
     8641 cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc
     8701 tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat atcctgggcc
     8761 ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa
     8821 atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg
     8881 ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc
     8941 actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg
     9001 ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaac
     9061 tcgagccaaa gaaaaagcgg aaagtgtaat ctagaggatc tgcccctctc cctccccccc
     9121 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     9181 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     9241 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     9301 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     9361 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     9421 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     9481 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aatgggctga aggatgccca
     9541 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     9601 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     9661 aaaaacacga tgataaggat ccaccggagc ttaccatgac cgagtacaag cccacggtgc
     9721 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     9781 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     9841 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     9901 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     9961 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
    10021 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
    10081 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
    10141 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
    10201 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
    10261 gcacctggtg catgacccgc aagcccggtg cctgacgccc atcgatagat cctaatcaac
    10321 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
    10381 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
    10441 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
    10501 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
    10561 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
    10621 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
    10681 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
    10741 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
    10801 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
    10861 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctgagatc ctttaagacc
    10921 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
    10981 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
    11041 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
    11101 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
    11161 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
    11221 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
    11281 gcccgggtta attaaggaaa gggctagatc attcttgaag acgaaagggc ctcgtgatac
    11341 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
    11401 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
    11461 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
    11521 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
    11581 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
    11641 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
    11701 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
    11761 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
    11821 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
    11881 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
    11941 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
    12001 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
    12061 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
    12121 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
    12181 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
    12241 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
    12301 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
    12361 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
    12421 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
    12481 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
    12541 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
    12601 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
    12661 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
    12721 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
    12781 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
    12841 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
    12901 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
    12961 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
    13021 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
    13081 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
    13141 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
    13201 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
    13261 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
    13321 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcaagc
    13381 tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
    13441 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccgt
    13501 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    13561 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    13621 gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt acgaatttca
    13681 caaataaagc attttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    13741 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
    13801 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
    13861 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
    13921 gtagt
//
