LentiGuide Cherry

LOCUS       LentiGuide Cherry               10277 bp ds-DNA     circular SYN 27-SEP-2021
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and mCherry marker from EF-1a promoter. 3rd 
            generation lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    LentiGuide Cherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10277)
  AUTHORS   Newman R, Tolar P
  TITLE     Genome-wide screens identify calcium signaling as a key regulator of
            IgE+ plasma cell differentiation and survival
  JOURNAL   biorXiv 2021.03.02.433398
REFERENCE   2  (bases 1 to 10277)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 27, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10277
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        87..1265
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1256
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1213..1233
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(1406..1424)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1674..1693
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1987..2575
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2040..2060)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2458..2469)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2647..2880
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2958..3079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2995..3014)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3049..3068
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3119..3254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3181..3200
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3274..3293)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3275..3293)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3303..3320)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3303..3319)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3312..3334)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3461..3916
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3548..3567)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3758..3779
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3942..4046
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4047..4907
                     /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(4265..4284)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5078..5666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5567..5586
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5820..5837
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5954..5975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5990..6020
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6028..6044
                     /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     6033..6055
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6052..6068
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6052..6068
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6087..6107
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6089..6107
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6135..6361
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6362..6542
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6589..6714
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7207..7440
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7625..7669
                     /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        7847..8087
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     7847..7867
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8018..8037
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9977..10052
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    10108..10225
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
      421 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      481 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      541 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      601 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      661 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      721 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      781 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      841 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      901 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      961 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     1021 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1081 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1141 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1201 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1261 cgtgacgtac gcctcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     1321 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     1381 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     1441 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     1501 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     1561 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     1621 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     1681 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     1741 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     1801 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     1861 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     1921 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta aacgcgttaa
     1981 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     2041 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     2101 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     2161 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     2221 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     2281 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     2341 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     2401 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     2461 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     2521 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
     2581 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     2641 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     2701 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     2761 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     2821 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     2881 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     2941 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3001 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3061 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     3121 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3181 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3241 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     3301 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3361 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3421 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     3481 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3541 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3601 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3661 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3721 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3781 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3841 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3901 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3961 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4021 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4081 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4141 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4201 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4261 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4321 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4381 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4441 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4501 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4561 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4621 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4681 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4741 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4801 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4861 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4921 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4981 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5041 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5101 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5161 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5221 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5281 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5341 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5401 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5461 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5521 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5581 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5641 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5701 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5761 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5821 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5881 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5941 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     6001 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6061 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     6121 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     6181 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     6241 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     6301 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     6361 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6421 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6481 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6541 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     6601 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     6661 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     6721 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     6781 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     6841 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     6901 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     6961 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     7021 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     7081 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     7141 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     7201 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     7261 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     7321 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     7381 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     7441 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     7501 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     7561 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     7621 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     7681 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     7741 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     7801 accattatcg tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc
     7861 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
     7921 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     7981 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     8041 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga
     8101 cggttgtaaa tgagcacaca aaatacacat gctaaaatat tatattctat gacctttata
     8161 aaatcaacca aaatcttctt tttaataact ttagtatcaa taattagaat ttttatgttc
     8221 ctttttgcaa acttttaata aaaatgagca aaataaaaaa acgctagttt tagtaactcg
     8281 cgttgttttc ttcaccttta ataatagcta ctccaccact tgttcctaag cggtcagctc
     8341 ctgcttcaat cattttttga gcatcttcaa atgttctaac tccaccagct gctttaacta
     8401 aagcattgtc tttaacaact gacttcatta gtttaacatc ttcaaatgtt gcacctgatt
     8461 ttgaaaatcc tgttgatgtt ttaacaaatt ctaatccagc ttcaacagct atttcacaag
     8521 ctttcatgat ttcttctttt gttaataaac aattttccat aatacattta acaacatgtg
     8581 atccagctgc tttttttaca gctttcatgt cttctaaaac taattcataa tttttgtctt
     8641 ttaatgcacc aatatttaat accatatcaa tttctgttgc accatcttta attgcttcag
     8701 aaacttcgaa tgcttttgta gctgttgtgc atgcacctag aggaaaacct acaacatttg
     8761 ttattcctac atttgtgcct tttaataatt ctttacaata gcttgttcaa tatgaattaa
     8821 cacaaactgt tgcaaaatca aattcaattg cttcatcaca taattgttta atttcagctt
     8881 tcgtagcatc ttgttttaat aatgtgtgat ctatatattt gtttagtttc attttttctc
     8941 ctatatattc atttttaatt ttaattcttt aataatttcg tctactttaa ctttagcgtt
     9001 ttgaacagat tcaccaacac ctataaaata aatttttagt ttaggttcag ttccacttgg
     9061 gcgaacagca aatcatgact tatcttctaa ataaaatttt agtaagtctt gtcctggcat
     9121 attatacatt ccatcgatgt agtcttcaac attaacaact ttaagtccag caatttgagt
     9181 taagggtgtt gctctcaatg atttcattaa tggttcaatt tttaatttct tttcttctgg
     9241 tttaaaattc aagtttaaag tgaaagtgta atatgcaccc atttctttaa ataaatcttc
     9301 taaatagtct actaatgttt tattttgttt tttataaaat caagcagcct ctgctattaa
     9361 tatagaagct tgtattccat ctttatctct agctgagtca tcaattacat atccataact
     9421 ttcttcataa gcaaaaacaa aatttaatcc gttatcttct tctttagcaa tttctctacc
     9481 cattcattta aatccagtta aagtttttac aatattaact ccatattttt catgagcgat
     9541 tctatcaccc aaatcacttg ttacaaaact tgaatataga gccggatttt ttggaatgct
     9601 atttaagcgt tttagatttg ataattttca atcaattaaa attggtcctg tttgatttcc
     9661 atctaatctt acaaaatgac catcatgttt tattgccatt ccaaatctgt cagcatctgg
     9721 gtcattcata ataataatat ctgcatcatg tttaatacca tattcaagcg gtatttttca
     9781 tgcaggatca aattctggat ttggatttac aacattttta aatgtttcat cttcaaatgc
     9841 atgctcttca acctcaataa cgttatatcc tgattcacgt aatatttttg gggtaaattt
     9901 agttcctgtt ccattaactg cgctaaaaat aatttttaaa tcttttttag cttcttgctc
     9961 ttttttgtac gtctctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
    10021 atcaacttga aaaagtggca ccgagtcggt gcttttttaa gcttggcgta actagatctt
    10081 gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
    10141 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
    10201 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg
    10261 gcgccggctc gaggggg
//