pLVX-MPro-eGFP-2

LOCUS       pLVX-MPro-eGFP-2              10493 bp ds-DNA     circular SYN 17-MAY-2022
DEFINITION  Expresses Mpro-eGFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pLVX-MPro-eGFP-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10493)
  AUTHORS   Cao W, Cho CD, Geng ZZ, Shaabani N, Ma XR, Vatansever EC, Alugubelli
            YR, Ma Y, Chaki SP, Ellenburg WH, Yang KS, Qiao Y, Allen R, Neuman 
            BW, Ji H, Xu S, Liu WR
  TITLE     Evaluation of SARS-CoV-2 Main Protease Inhibitors Using a Novel 
            Cell-Based Assay.
  JOURNAL   ACS Cent Sci. 2022 Feb 23;8(2):192-204. doi: 
            10.1021/acscentsci.1c00910. Epub 2022 Feb 2.
  PUBMED    35229034
REFERENCE   2  (bases 1 to 10493)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 17, 2022 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10493
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             32..665
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    712..837
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1334..1567
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1752..1796
                     /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    2059..2174
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2369..3550
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2599..3541
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3498..3518
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4538..5257
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             4538..5254
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4583..4604)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4844..4863)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5191..5212
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5274..5847
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5441..5458)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5668..5687
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5880..6479
                     /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(5880..5899)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6376..6396
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6493..7081
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6546..6566)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6964..6975)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7288..7921
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8049..8065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8049..8065)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8062..8084)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8073..8089
                     /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(8097..8127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8142..8163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8280..8297)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8451..9039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8531..8550)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9210..10070)
                     /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     9833..9852
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10071..10175)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    10223..10357
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10260..10279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10314..10333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga
       61 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac
      121 tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta
      181 ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac
      241 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac
      301 agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc
      361 tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
      421 ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg
      481 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
      541 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
      601 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
      661 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
      721 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
      781 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
      841 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
      901 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
      961 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     1021 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     1081 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     1141 gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg
     1201 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     1261 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     1321 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     1381 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     1441 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     1501 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     1561 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     1621 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     1681 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     1741 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     1801 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     1861 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     1921 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     1981 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     2041 gatctcgacg gtatcgcctt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagttta tcgatgagta
     2221 attcatacaa aaggactcgc ccctgccttg gggaatccca gggaccgtcg ttaaactccc
     2281 actaacgtag aacccagaga tcgctgcgtt cccgccccct cacccgcccg ctctcgtcat
     2341 cactgaggtg gagaagagca tgcgtgaggc tccggtgccc gtcagtgggc agagcgcaca
     2401 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     2461 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2521 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2581 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg
     2641 ggttatggcc cttgcgtgcc ttgaattact tccacgcccc tggctgcagt acgtgattct
     2701 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
     2761 cccttcgcct cgtgcttgag ttgaggcctg gcttgggcgc tggggccgcc gcgtgcgaat
     2821 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
     2881 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
     2941 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
     3001 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
     3061 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
     3121 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
     3181 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     3241 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     3301 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     3361 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     3421 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     3481 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     3541 ggtgtcgtga ggatctattt ccggtgaatt cctcgagact agttctagaa tgaaaacaag
     3601 cgcggtgctc cagtcagggt ttcgcaagat ggcctttcca agcggtaagg ttgaaggttg
     3661 catggtacaa gtaacgtgcg gtacgactac gttgaacggc ctgtggttgg acgatgtggt
     3721 atattgcccc cgccacgtca tctgcacctc cgaagacatg ttaaacccca attacgagga
     3781 tttgcttatc cgtaaaagca accataactt ccttgtgcaa gcaggcaatg tgcaattacg
     3841 tgttattggc cactccatgc agaattgcgt acttaaatta aaggtggata cggccaaccc
     3901 taaaacgccc aaatataaat ttgtccgcat tcaaccgggt caaaccttct ctgtattggc
     3961 ctgttacaac ggttccccca gtggtgttta tcagtgtgca atgcgcccga acttcacaat
     4021 caagggatcg ttcctgaatg ggagttgtgg ttcggttgga ttcaatatcg actacgactg
     4081 cgtctctttt tgctatatgc accatatgga attaccgacc ggcgtacatg ccgggaccga
     4141 tcttgaagga aatttctacg gcccgtttgt cgatcgtcag actgctcaag cagcaggtac
     4201 ggatacgaca attaccgtga acgtccttgc atggctgtat gccgcggtca ttaacggaga
     4261 tcgttggttt cttaaccgtt ttactaccac attaaacgat ttcaacttgg tggccatgaa
     4321 gtacaattat gagcctctta cgcaagatca tgtagatatt ctgggacctt tatcggccca
     4381 aacaggaatc gctgtgctgg acatgtgcgc atcattgaaa gagttgctgc agaatggtat
     4441 gaacggacgc acaattctgg ggtcggcgtt gctggaggat gaattcaccc cgtttgatgt
     4501 tgtgcgtcag tgttccggcg taacttttgg caagcttatg gtgagcaagg gcgaggagct
     4561 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     4621 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     4681 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     4741 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     4801 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     4861 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     4921 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     4981 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     5041 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     5101 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     5161 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     5221 cgggatcact ctcggcatgg acgagctgta caagtaagcg gccgcggatc ccgcccctct
     5281 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     5341 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     5401 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     5461 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     5521 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     5581 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     5641 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     5701 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     5761 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     5821 gttttccttt gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca
     5881 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     5941 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     6001 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     6061 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     6121 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     6181 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     6241 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     6301 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     6361 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     6421 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctaga
     6481 cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     6541 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6601 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6661 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6721 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6781 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6841 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     6901 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6961 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     7021 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     7081 cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata
     7141 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     7201 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     7261 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
     7321 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     7381 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     7441 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     7501 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     7561 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     7621 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     7681 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     7741 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7801 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7861 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     7921 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     7981 gagagtgaga ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg
     8041 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     8101 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     8161 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     8221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     8281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     8341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     8401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     8461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     8521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     8581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     8641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     8821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     8881 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     8941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     9001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     9061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     9121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     9181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     9241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     9301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     9361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     9421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     9481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     9541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     9601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     9841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     9901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     9961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    10021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    10081 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    10141 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    10201 gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag
    10261 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    10321 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
    10381 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
    10441 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gta
//