LOCUS       sequence-330062-       11028 bp    DNA     circular SYN 14-APR-2025
DEFINITION  3rd generation lentiviral expression of C-myc, CDK4(R24C), and GFP 
            separated by 2A self-cleaving peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11028)
  TITLE     Brooks lab unpublished viral plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11028)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLenti-PGK-c-myc-2A-CDK4(R24C)-GFP-Puro
FEATURES             Location/Qualifiers
     source          1..11028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    226..351
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    844..1077
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1262..1306
                     /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    1573..1690
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1736..2246
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2165..2183
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    2301..2325
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2326..3642
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     CDS             3643..3696
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4618..4671
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4684..4686,4687..4689,4690..5403)
                     /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             join(4684..4686,4687..4689,4690..5400)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4729..4750)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4990..5009)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5337..5358
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5404..5428)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5459..6047
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5512..5532)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5930..5941)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        6061..6560
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6096..6119)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6493..6512
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6581..7180
                     /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(6581..6600)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7077..7097
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7318..7551
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7623..7744
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7660..7679)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7714..7733
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7784..7919
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7939..7958)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7940..7958)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7968..7985)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7968..7984)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7977..7999)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8126..8581
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8213..8232)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8423..8444
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8607..8711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8712..8780,8781..9572)
                     /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     complement(8930..8949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9743..10331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10232..10251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10485..10502
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10619..10640
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10655..10660,10661..10678,10679..10685)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10693..10709
                     /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     10698..10720
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10717..10733
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10717..10733
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10752..10772
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10754..10772
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10800..11026
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             join(11027..11028,1..179)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
       61 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
      121 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
      181 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
      241 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
      301 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
      361 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
      421 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
      481 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
      541 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
      601 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
      661 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
      721 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
      781 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
      841 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
      901 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
      961 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1021 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1081 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1141 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     1201 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     1261 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     1321 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     1381 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     1441 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     1501 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg
     1561 gtatcggtta acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     1621 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     1681 tcaaaatttt atcgatcacg agactagcct cgagaagctt gatatcgaat tccacggggt
     1741 tggggttgcg ccttttccaa ggcagccctg ggtttgcgca gggacgcggc tgctctgggc
     1801 gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc acgtccgttc
     1861 gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc cccggcgacg cttcctgctc
     1921 cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga caaacggaag
     1981 ccgcacgtct cactagtacc ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc
     2041 gaccgcgatg ggctgtggcc aatagcggct gctcagcagg gcgcgccgag agcagcggcc
     2101 gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg
     2161 cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg
     2221 accgaatcac cgacctctct ccccaggggg atccaccggt tcgtcgacta gtccagtgtg
     2281 gtggaattct gcagatatca acaagtttgt acaaaaaagc aggctatgcc cctcaacgtt
     2341 agcttcacca acaggaacta tgacctcgac tacgactcgg tgcagccgta tttctactgc
     2401 gacgaggagg agaacttcta ccagcagcag cagcagagcg agctgcagcc cccggcgccc
     2461 agcgaggata tctggaagaa attcgagctg ctgcccaccc cgcccctgtc ccctagccgc
     2521 cgctccgggc tctgctcgcc ctcctacgtt gcggtcacac ccttctccct tcggggagac
     2581 aacgacggcg gtggcgggag cttctccacg gccgaccagc tggagatggt gaccgagctg
     2641 ctgggaggag acatggtgaa ccagagtttc atctgcgacc cggacgacga gaccttcatc
     2701 aaaaacatca tcatccagga ctgtatgtgg agcggcttct cggccgccgc caagctcgtc
     2761 tcagagaagc tggcctccta ccaggctgcg cgcaaagaca gcggcagccc gaaccccgcc
     2821 cgcggccaca gcgtctgctc cacctccagc ttgtacctgc aggatctgag cgccgccgcc
     2881 tcagagtgca tcgacccctc ggtggtcttc ccctaccctc tcaacgacag cagctcgccc
     2941 aagtcctgcg cctcgcaaga ctccagcgcc ttctctccgt cctcggattc tctgctctcc
     3001 tcgacggagt cctccccgca gggcagcccc gagcccctgg tgctccatga ggagacaccg
     3061 cccaccacca gcagcgactc tgaggaggaa caagaagatg aggaagaaat cgatgttgtt
     3121 tctgtggaaa agaggcaggc tcctggcaaa aggtcagagt ctggatcacc ttctgctgga
     3181 ggccacagca aacctcctca cagcccactg gtcctcaaga ggtgccacgt ctccacacat
     3241 cagcacaact acgcagcgcc tccctccact cggaaggact atcctgctgc caagagggtc
     3301 aagttggaca gtgtcagagt cctgagacag atcagcaaca accgaaaatg caccagcccc
     3361 aggtcctcgg acaccgagga gaatgtcaag aggcgaacac acaacgtctt ggagcgccag
     3421 aggaggaacg agctaaaacg gagctttttt gccctgcgtg accagatccc ggagttggaa
     3481 aacaatgaaa aggcccccaa ggtagttatc cttaaaaaag ccacagcata catcctgtcc
     3541 gtccaagcag aggagcaaaa gctcatttct gaagaggact tgttgcggaa acgacgagaa
     3601 cagttgaaac acaaacttga acagctacgg aactcttgtg cggagggcag gggaagcctg
     3661 ctcacctgcg gcgacgtgga ggaaaaccct ggccccgccc caggatccat ggctacctct
     3721 cgatatgagc cagtggctga aattggtgtc ggtgcctatg ggacagtgta caaagcttgt
     3781 gatccccaca gtggccactt tgtggccctc aagagtgtga gagtccccaa tggaggagga
     3841 ggtggaggag gccttcccat cagcacagtt cgtgaggtgg ctttactgag gcgactggag
     3901 gcttttgagc atcccaatgt tgtccggctg atggacgtct gtgccacatc ccgaactgac
     3961 cgggagatca aggtaaccct ggtgtttgag catgtagacc aggacctaag gacatatctg
     4021 gacaaggcac ccccaccagg cttgccagcc gaaacgatca aggatctgat gcgccagttt
     4081 ctaagaggcc tagatttcct tcatgccaat tgcatcgttc accgagatct gaagccagag
     4141 aacattctgg tgacaagtgg tggaacagtc aagctggctg actttggcct ggccagaatc
     4201 tacagctacc agatggcact tacacccgtg gttgttacac tctggtaccg agctcccgaa
     4261 gttcttctgc agtccacata tgcaacacct gtggacatgt ggagtgttgg ctgtatcttt
     4321 gcagagatgt ttcgtcgaaa gcctctcttc tgtggaaact ctgaagccga ccagttgggc
     4381 aaaatctttg acctgattgg gctgcctcca gaggatgact ggcctcgaga tgtatccctg
     4441 ccccgtggag cctttccccc cagagggccc cgcccagtgc agtcggtggt acctgagatg
     4501 gaggagtcgg gagcacagct gctgctggaa atgctgactt ttaacccaca caagcgaatc
     4561 tctgcctttc gagctctgca gcactcttat ctacataagg atgaaggtaa tccggaggag
     4621 ggcaggggaa gcctcctgac atgtggagat gtcgaagaga atcccggccc tgccccagga
     4681 tccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4741 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4801 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4861 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     4921 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     4981 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5041 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     5101 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     5161 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     5221 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     5281 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     5341 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5401 taaacccagc tttcttgtac aaagtggttg atatccagca cagtggcggc cgctcgacaa
     5461 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     5521 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     5581 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     5641 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5701 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     5761 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5821 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     5881 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5941 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     6001 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattctacc
     6061 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     6121 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     6181 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     6241 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     6301 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     6361 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     6421 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     6481 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     6541 cctcatctcc gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac
     6601 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     6661 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     6721 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     6781 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     6841 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     6901 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     6961 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7021 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     7081 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     7141 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca
     7201 gcgcccgacc gaaaggagcg cacgacccca tgcatctcga gggcccggta cctttaagac
     7261 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     7321 aagggctagc tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     7381 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7441 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7501 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     7561 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     7621 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7681 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7741 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     7801 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     7861 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     7921 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     7981 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     8041 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     8101 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     8161 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     8221 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     8281 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     8341 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     8401 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     8461 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     8521 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     8581 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     8641 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     8701 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     8761 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     8821 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     8881 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     8941 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     9001 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     9061 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     9121 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     9181 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     9241 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     9301 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     9361 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     9421 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     9481 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     9541 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     9601 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     9661 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     9721 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     9781 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     9841 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     9901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     9961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
    10021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
    10081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
    10141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
    10201 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
    10261 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
    10321 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
    10381 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
    10441 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
    10501 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
    10561 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
    10621 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
    10681 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
    10741 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
    10801 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
    10861 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
    10921 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
    10981 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgg
//
