LOCUS       sequence-361388-       10583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses guide RNA, contains intact LTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10583)
  AUTHORS   Iaffaldano BJ, Marino MP, Reiser J
  TITLE     CRISPR library screening to develop HEK293-derived cell lines with 
            improved lentiviral vector titers.
  JOURNAL   Front Genome Ed. 2023 Jul 13;5:1218328. doi: 
            10.3389/fgeed.2023.1218328. eCollection 2023.
  PUBMED    37520398
REFERENCE   2  (bases 1 to 10583)
  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: "Front 
            Genome Ed."; date: "2023-07-13"; volume: "5:1218328. doi"; pages: " 
            10.3389/fgeed.2023.1218328. eCollection 2023"
COMMENT     Sequence Label: lentiguide-puro-wt-ltr
FEATURES             Location/Qualifiers
     source          1..10583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        191..439
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     191..211
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     362..381
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2321..2396
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2452..2569
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2708..3886
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2939..3877
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3834..3854
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3899..4498
                     /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(3899..3918)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4395..4415
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4514..5102
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4567..4587)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4985..4996)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5174..5807
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5885..6006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5922..5941)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5976..5995
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6046..6181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6201..6220)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6202..6220)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6230..6247)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6230..6246)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6239..6261)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6388..6843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6475..6494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6685..6706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6869..6973
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6974..7042,7043..7834)
                     /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(7192..7211)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8005..8593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8494..8513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8747..8764
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8881..8902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8917..8922,8923..8940,8941..8947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8955..8971
                     /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     8960..8982
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8979..8995
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8979..8995
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9014..9034
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9016..9034
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9062..9288
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9289..9469
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9516..9641
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10134..10367
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             join(10552..10583,1..13)
                     /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"
ORIGIN
        1 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
       61 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
      121 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
      181 gggacccaga gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
      241 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
      301 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
      361 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
      421 gtggaaagga cgaaacaccg gagacggttg taaatgagca cacaaaatac acatgctaaa
      481 atattatatt ctatgacctt tataaaatca accaaaatct tctttttaat aactttagta
      541 tcaataatta gaatttttat gttccttttt gcaaactttt aataaaaatg agcaaaataa
      601 aaaaacgcta gttttagtaa ctcgcgttgt tttcttcacc tttaataata gctactccac
      661 cacttgttcc taagcggtca gctcctgctt caatcatttt ttgagcatct tcaaatgttc
      721 taactccacc agctgcttta actaaagcat tgtctttaac aactgacttc attagtttaa
      781 catcttcaaa tgttgcacct gattttgaaa atcctgttga tgttttaaca aattctaatc
      841 cagcttcaac agctatttca caagctttca tgatttcttc ttttgttaat aaacaatttt
      901 ccataataca tttaacaaca tgtgatccag ctgctttttt tacagctttc atgtcttcta
      961 aaactaattc ataatttttg tcttttaatg caccaatatt taataccata tcaatttctg
     1021 ttgcaccatc tttaattgct tcagaaactt cgaatgcttt tgtagctgtt gtgcatgcac
     1081 ctagaggaaa acctacaaca tttgttattc ctacatttgt gccttttaat aattctttac
     1141 aatagcttgt tcaatatgaa ttaacacaaa ctgttgcaaa atcaaattca attgcttcat
     1201 cacataattg tttaatttca gctttcgtag catcttgttt taataatgtg tgatctatat
     1261 atttgtttag tttcattttt tctcctatat attcattttt aattttaatt ctttaataat
     1321 ttcgtctact ttaactttag cgttttgaac agattcacca acacctataa aataaatttt
     1381 tagtttaggt tcagttccac ttgggcgaac agcaaatcat gacttatctt ctaaataaaa
     1441 ttttagtaag tcttgtcctg gcatattata cattccatcg atgtagtctt caacattaac
     1501 aactttaagt ccagcaattt gagttaaggg tgttgctctc aatgatttca ttaatggttc
     1561 aatttttaat ttcttttctt ctggtttaaa attcaagttt aaagtgaaag tgtaatatgc
     1621 acccatttct ttaaataaat cttctaaata gtctactaat gttttatttt gttttttata
     1681 aaatcaagca gcctctgcta ttaatataga agcttgtatt ccatctttat ctctagctga
     1741 gtcatcaatt acatatccat aactttcttc ataagcaaaa acaaaattta atccgttatc
     1801 ttcttcttta gcaatttctc tacccattca tttaaatcca gttaaagttt ttacaatatt
     1861 aactccatat ttttcatgag cgattctatc acccaaatca cttgttacaa aacttgaata
     1921 tagagccgga ttttttggaa tgctatttaa gcgttttaga tttgataatt ttcaatcaat
     1981 taaaattggt cctgtttgat ttccatctaa tcttacaaaa tgaccatcat gttttattgc
     2041 cattccaaat ctgtcagcat ctgggtcatt cataataata atatctgcat catgtttaat
     2101 accatattca agcggtattt ttcatgcagg atcaaattct ggatttggat ttacaacatt
     2161 tttaaatgtt tcatcttcaa atgcatgctc ttcaacctca ataacgttat atcctgattc
     2221 acgtaatatt tttggggtaa atttagttcc tgttccatta actgcgctaa aaataatttt
     2281 taaatctttt ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc
     2341 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     2401 ttaagcttgg cgtaactaga tcttgagaca aatggcagta ttcatccaca attttaaaag
     2461 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2521 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     2581 cagggacagc agagatccac tttggcgccg gctcgagggg gcccgggtgc aaagatggat
     2641 aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt
     2701 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     2761 gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg
     2821 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     2881 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
     2941 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc
     3001 ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
     3061 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
     3121 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
     3181 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
     3241 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
     3301 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
     3361 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
     3421 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     3481 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
     3541 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     3601 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
     3661 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     3721 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     3781 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     3841 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta cggccaccat
     3901 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     3961 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     4021 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     4081 cggcaaggtg tgggtcgcgg acgacggcgc cgccgtggcg gtctggacca cgccggagag
     4141 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     4201 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     4261 cgcgtggttc ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag
     4321 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     4381 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     4441 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac
     4501 gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4561 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4621 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4681 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4741 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4801 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4861 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     4921 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4981 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5041 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5101 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     5161 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     5221 tggatctacc acacacaagg ctacttccct gattggcaga actacacacc agggccaggg
     5281 atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag
     5341 gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg
     5401 atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat
     5461 cacatggccc gagagctgca tccggagtac ttcaagaact gctgacatcg agcttgctac
     5521 aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg
     5581 cgagccctca gatgctgcat ataagcagct gctttttgct tgtactgggt ctctctggtt
     5641 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     5701 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     5761 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag
     5821 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     5881 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5941 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6001 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact
     6061 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6121 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6181 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     6241 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     6301 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     6361 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     6421 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6481 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6541 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6601 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     6661 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6721 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     6781 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     6841 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6901 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6961 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7021 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7081 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7141 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7201 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7261 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7321 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7381 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7441 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7501 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7561 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7621 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7681 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7741 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7801 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7861 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7921 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7981 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8041 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8101 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8161 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8221 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8281 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8341 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8401 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8461 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8521 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8581 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     8641 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     8701 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     8761 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     8821 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     8881 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     8941 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     9001 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta
     9061 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
     9121 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
     9181 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
     9241 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
     9301 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     9361 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     9421 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     9481 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     9541 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     9601 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     9661 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     9721 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     9781 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     9841 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     9901 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     9961 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
    10021 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
    10081 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
    10141 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
    10201 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
    10261 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
    10321 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
    10381 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
    10441 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
    10501 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
    10561 caagaattat tggaattaga taa
//
