LOCUS       sequence-237089-       10538 bp    DNA     circular SYN 14-APR-2025
DEFINITION  2nd generation lentiviral transfer plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10538)
  TITLE     Trono Lab Constitutive Lentiviral Plasmids
REFERENCE   2  (bases 1 to 10538)
  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"
COMMENT     Sequence Label: pWPXL
FEATURES             Location/Qualifiers
     source          1..10538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             2..635
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    682..807
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1300..1533
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1718..1762
                     /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"
     promoter        2145..3323
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2376..3314
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3271..3291
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    3371..3488
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      3565..3574
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3571..3573,3574..3576,3577..4287)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3571..3573,3574..3576,3577..4287)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3616..3637)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3877..3896)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4224..4245
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4369..4957
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4422..4442)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4840..4851)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(5101..5134)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5154..5334
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(5359..5377)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5360..5377)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5667..5686)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(5900..5919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6019..6041
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6079..6097)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6165..6269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6270..6338,6339..7130)
                     /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(6488..6507)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7301..7889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7790..7809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8043..8060
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8130..8150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8135..8464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8315..8450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          9598..9663
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9793..9813
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10238..10372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10275..10294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10329..10348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg gatctaccac
       61 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
      121 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
      181 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
      241 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
      301 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
      361 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
      421 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
      481 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      541 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      601 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      661 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      721 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      781 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      841 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      901 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
      961 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1021 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1081 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1141 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1201 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1261 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1321 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1381 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1441 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1501 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1561 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1621 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1681 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1741 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1801 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1861 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1921 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     1981 agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg
     2041 accatcgatg tcgacgataa gctttgcaaa gatggataaa gttttaaaca gagaggaatc
     2101 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
     2161 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     2221 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     2281 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     2341 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     2401 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
     2461 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
     2521 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
     2581 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     2641 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     2701 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     2761 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     2821 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     2881 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     2941 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     3001 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     3061 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     3121 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     3181 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     3241 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     3301 ttcttccatt tcaggtgtcg tgaggaattt cgacatttaa atttaattaa tctcgacggt
     3361 atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     3421 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     3481 aaaattttat cgatcacgag actagcctcg aggtttaaac tacgggatcc aggcctaagc
     3541 ttacgcgtcc tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
     3601 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3661 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3721 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     3781 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3841 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3901 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3961 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     4021 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     4081 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     4141 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     4201 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4261 actctcggca tggacgagct gtacaagtcc ggactcagat ctcgactagc tagtagctag
     4321 ctagctagtc gagctcaagc ttcgaattcc cgggactagt catatgataa tcaacctctg
     4381 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4441 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     4501 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     4561 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     4621 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     4681 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     4741 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     4801 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     4861 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     4921 cagacgagtc ggatctccct ttgggccgcc tccccgcatc ggtacgtatg gccaggtacc
     4981 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     5041 gggactggaa gggctaattc actcccaacg aagacaagat gggatcaatt caccatggga
     5101 ataacttcgt atagcataca ttatacgaag ttatgctgct ttttgcttgt actgggtctc
     5161 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     5221 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     5281 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct
     5341 agaattaatt ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt
     5401 tatgtattaa ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc
     5461 tggcttactg aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt
     5521 tggacgaacc ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc
     5581 aatcagcagg gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac
     5641 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     5701 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     5761 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     5821 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     5881 tcgaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     5941 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     6001 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     6061 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     6121 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6181 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6241 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6301 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6361 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6421 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6481 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6541 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     6601 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6661 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     6721 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6781 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6841 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6901 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6961 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7021 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7081 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7141 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7201 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7261 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7321 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7381 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7441 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7501 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7561 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7621 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7681 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7741 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7801 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7861 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7921 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7981 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     8041 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     8101 cgcgttggcc gattcattaa tgcagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     8161 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     8221 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     8281 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     8341 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     8401 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     8461 caaaaagctt ggacacaaga caggcttgcg agatatgttt gagaatacca ctttatcccg
     8521 cgtcagggag aggcagtgcg taaaaagacg cggactcatg tgaaatactg gtttttagtg
     8581 cgccagatct ctataatctc gcgcaaccta ttttcccctc gaacactttt taagccgtag
     8641 ataaacaggc tgggacactt cacatgagcg aaaaatacat cgtcacctgg gacatgttgc
     8701 agatccatgc acgtaaactc gcaagccgac tgatgccttc tgaacaatgg aaaggcatta
     8761 ttgccgtaag ccgtggcggt ctgtaccggg tgcgttactg gcgcgtgaac tgggtattcg
     8821 tcatgtcgat accgtttgta tttccagcta cgatcacgac aaccagcgcg agcttaaagt
     8881 gctgaaacgc gcagaaggcg atggcgaagg cttcatcgtt attgatgacc tggtggatac
     8941 cggtggtact gcggttgcga ttcgtgaaat gtatccaaaa gcgcactttg tcaccatctt
     9001 cgcaaaaccg gctggtcgtc cgctggttga tgactatgtt gttgatatcc cgcaagatac
     9061 ctggattgaa cagccgtggg atatgggcgt cgtattcgtc ccgccaatct ccggtcgcta
     9121 atcttttcaa cgcctggcac tgccgggcgt tgttcttttt aacttcaggc gggttacaat
     9181 agtttccagt aagtattctg gaggctgcat ccatgacaca ggcaaacctg agcgaaaccc
     9241 tgttcaaacc ccgctttaaa catcctgaaa cctcgacgct agtccgccgc tttaatcacg
     9301 gcgcacaacc gcctgtgcag tcggcccttg atggtaaaac catccctcac tggtatcgca
     9361 tgattaaccg tctgatgtgg atctggcgcg gcattgaccc acgcgaaatc ctcgacgtcc
     9421 aggcacgtat tgtgatgagc gatgccgaac gtaccgacga tgatttatac gatacggtga
     9481 ttggctaccg tggcggcaac tggatttatg agtgggcccc ggatctttgt gaaggaacct
     9541 tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta
     9601 aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt
     9661 tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa
     9721 aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa
     9781 cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa
     9841 ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt
     9901 tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta
     9961 acctttataa gtaggcataa cagttataat cataacatac tgttttttct tactccacac
    10021 aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta
    10081 atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag agatcataat
    10141 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
    10201 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
    10261 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
    10321 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
    10381 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
    10441 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
    10501 aaattgtatt tgttaaatat gtactacaaa cttagtag
//
