LOCUS       sequence-226380-       10040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for CRISPR mediated gene editing in cells while 
            expressing Firefly Luciferase for in vivo imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10040)
  AUTHORS   Chow RD, Guzman CD, Wang G, Schmidt F, Youngblood MW, Ye L, Errami 
            Y, Dong MB, Martinez MA, Zhang S, Renauer P, Bilguvar K, Gunel M, 
            Sharp PA, Zhang F, Platt RJ, Chen S
  TITLE     AAV-mediated direct in vivo CRISPR screen identifies functional 
            suppressors in glioblastoma.
  JOURNAL   Nat Neurosci. 2017 Aug 14. doi: 10.1038/nn.4620.
  PUBMED    28805815
REFERENCE   2  (bases 1 to 10040)
  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: "Nat 
            Neurosci. 2017 Aug 14. doi: 10.1038/nn.4620."
COMMENT     Sequence Label: pLenti-U6sgbbBsmbI-puro-2A-Fluc
FEATURES             Location/Qualifiers
     source          1..10040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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        1717..1965
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1717..1737
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1888..1907
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1982..2057
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2113..2230
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2369..3547
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2600..3538
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3495..3515
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3560..4156
                     /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(3560..3579)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4056..4076
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4166..4222
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4223..5875
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(4289..4307)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    5897..6485
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5950..5970)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6368..6379)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6557..6790
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6868..6989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6905..6924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6959..6978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7029..7164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7184..7203)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7185..7203)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7213..7230)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7213..7229)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7222..7244)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7371..7826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7458..7477)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7668..7689
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7852..7956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7957..8025,8026..8817)
                     /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(8175..8194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8988..9576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9477..9496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9730..9747
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9864..9885
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9900..9905,9906..9923,9924..9930)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9938..9954
                     /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     9943..9965
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9962..9978
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9962..9978
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9997..10017
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9999..10017
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct
     1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga cggacgtctc
     1981 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
     2041 tggcaccgag tcggtgcttt tttaagcttg gcgtaactag atcttgagac aaatggcagt
     2101 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     2161 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     2221 tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggcgcc ggctcgaggg
     2281 ggcccgggtg caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac
     2341 cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     2401 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     2461 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2521 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2581 gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac
     2641 gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg tgattcttga
     2701 tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc
     2761 ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg
     2821 gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg
     2881 atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct
     2941 gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg
     3001 cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc
     3061 tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg
     3121 ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg
     3181 ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc
     3241 acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga
     3301 gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt
     3361 aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga
     3421 agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg
     3481 atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt
     3541 gtcgtgacgt acggccacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     3601 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     3661 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     3721 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgccgtggc
     3781 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     3841 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     3901 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca
     3961 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     4021 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     4081 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     4141 ccgcaagccc ggtgccggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga
     4201 tgtcgaagag aatcctggac cgatggaaga cgccaaaaac ataaagaaag gcccggcgcc
     4261 attctatccg ctggaagatg gaaccgctgg agagcaactg cataaggcta tgaagagata
     4321 cgccctggtt cctggaacaa ttgcttttac agatgcacat atcgaggtgg acatcactta
     4381 cgctgagtac ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa
     4441 tacaaatcac agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt
     4501 gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac gacatttata atgaacgtga
     4561 attgctcaac agtatgggca tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt
     4621 gcaaaaaatt ttgaacgtgc aaaaaaagct cccaatcatc caaaaaatta ttatcatgga
     4681 ttctaaaacg gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc
     4741 tcccggtttt aatgaatacg attttgtgcc agagtccttc gatagggaca agacaattgc
     4801 actgatcatg aactcctctg gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag
     4861 aactgcctgc gtgagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc
     4921 ggatactgcg attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact
     4981 cggatatttg atatgtggat ttcgagtcgt cttaatgtat agatttgaag aggagctgtt
     5041 tctgaggagc cttcaggatt acaagattca aagtgcgctg ctggtgccaa ccctattctc
     5101 cttcttcgcc aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc
     5161 ttctggtggc gctcccctct ctaaggaagt cggggaagcg gttgccaaga ggttccatct
     5221 gccaggtatc aggcaaggat atgggctcac tgagactaca tcagctattc tgattacacc
     5281 cgagggggat gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt
     5341 tgtggatctg gataccggga aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag
     5401 aggtcctatg attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga
     5461 caaggatgga tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt
     5521 catcgttgac cgcctgaagt ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga
     5581 attggaatcc atcttgctcc aacaccccaa catcttcgac gcaggtgtcg caggtcttcc
     5641 cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat
     5701 gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg
     5761 cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag
     5821 aaaaatcaga gagatcctca taaaggccaa gaagggcgga aagatcgccg tgtaagaatt
     5881 cacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5941 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     6001 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6061 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6121 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6181 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6241 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     6301 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6361 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6421 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6481 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6541 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     6601 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6661 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6721 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6781 atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca
     6841 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6901 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6961 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     7021 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     7081 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     7141 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     7201 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     7261 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     7321 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     7381 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     7441 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     7501 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     7561 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     7621 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     7681 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     7741 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     7801 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     7861 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7921 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7981 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     8041 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     8101 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     8161 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     8221 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     8281 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     8341 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     8401 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     8461 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     8521 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     8581 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     8641 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     8701 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8761 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8821 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8881 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8941 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     9001 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     9061 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     9121 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     9181 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     9241 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     9301 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     9361 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     9421 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     9481 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     9541 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     9601 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     9661 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     9721 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     9781 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     9841 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     9901 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     9961 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
    10021 caaaagctgg agctgcaagc
//
