LOCUS       sequence-173968-        8431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral CRISPR system backbone bearing BsmBI site for S. pyogenes
            new guide RNAs and TdTomato..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8431)
  AUTHORS   Singer M, Wang C, Cong L, Marjanovic ND, Kowalczyk MS, Zhang H, 
            Nyman J, Sakuishi K, Kurtulus S, Gennert D, Xia J, Kwon JY, Nevin J,
            Herbst RH, Yanai I, Rozenblatt-Rosen O, Kuchroo VK, Regev A, 
            Anderson AC
  TITLE     A Distinct Gene Module for Dysfunction Uncoupled from Activation in 
            Tumor-Infiltrating T Cells.
  JOURNAL   Cell. 2016 Sep 8;166(6):1500-1511.e9. doi: 
            10.1016/j.cell.2016.08.052.
  PUBMED    27610572
REFERENCE   2  (bases 1 to 8431)
  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"; doi: 
            "10.1016/j.cell.2016.08"; journalName: "Cell"; date: "2016-09-8- 8";
            volume: "166"; issue: "6"; pages: "1500-1511.e9"
COMMENT     Sequence Label: pCK005_U6-Sp-sgRNA_UP-TdTomato_WPRE
FEATURES             Location/Qualifiers
     source          1..8431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1132..1249
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(1305..1380)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(1397..1645)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(1455..1474)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1625..1645)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(1823..1867)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2052..2285
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2778..2903
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2950..3130
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(3131..3357)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(3385..3405)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3385..3403)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3424..3440)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3424..3440)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3437..3459)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3448..3464
                     /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)."
     promoter        complement(join(3472..3478,3479..3496,3497..3502))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3517..3538
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3655..3672)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3826..4414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3906..3925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4585..5376,5377..5445))
                     /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     5208..5227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5446..5550)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5576..6031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5713..5734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5925..5944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6158..6180
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6172..6189
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6173..6189
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6199..6218
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6199..6217
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      6238..6373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6413..6534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6424..6443)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6478..6497
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             6612..6845
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    6917..7505
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7023..7034
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7432..7452
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7530..8231)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(7613..7636)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7811..7830)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8080..8098
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      8228..8237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(join(8273..8431,1..1053))
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(join(8417..8431,1..5))
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 cttcagctac ttaagaagat aggtacataa aaccgaccaa agaaactgac gcctcactta
       61 tccctcccct caccagaggt ccggcgcctg tcgattcagg agagcctacc ctaggcccga
      121 accctgcgtc ctgcgacgga gaaaagccta ccgcacacct accggcaggt ggccccaccc
      181 tgcattataa gccaacagaa cgggtgacgt cacgacacga cgagggcgcg cgctcccaaa
      241 ggtacgggtg cactgcccaa cggcaccgcc ataactgccg cccccgcaac agacgacaaa
      301 ccgagttctc cagtcagtga caaacttcac gtcagggtcc ccagatggtg ccccagccca
      361 tctcacccga ataagagctt tcccgcatta gcgaaggcct caagaccttg ggttcttgcc
      421 gcccaccatg ccccccacct tgtttcaacg acctcacagc ccgcctcaca agcgtcttcc
      481 attcaagact cgggaacagc cgccattttg ctgcgctccc cccaaccccc agttcagggc
      541 aaccttgctc gcggacccag actacagccc ttggcggtct ctccacacgc ttccgtccca
      601 ccgagcggcc cggcggccac gaaagccccg gccagcccag cagcccgcta ctcaccaagt
      661 gacgatcaca gcgatccaca aacaagaacc gcgacccaaa tcccggctgc gacggaacta
      721 gctgtgccac acccggcgcg tccttatata atcatcggcg ttcaccgccc cacggagatc
      781 cctccgcaga atcgccgaga agggactact tttcctcgcc tgttccgctc tctggaaaga
      841 aaaccagtgc cctagagtca cccaagtccc gtcctaaaat gtccttctgc tgatactggg
      901 gttctaaggc cgagtcttat gagcagcggg ccgctgtcct gagcgtccgg gcggaaggat
      961 caggacgctc gctgcgccct tcgtctgacg tggcagcgct cgccgtgagg aggggggcgc
     1021 ccgcgggagg cgccaaaacc cggcgcggag gccagatctg gagccgacac gggttagggc
     1081 cccctcgagc cggcgccaaa gtggatctct gctgtccctg taataaaccc gaaaattttg
     1141 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
     1201 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa
     1261 tactgccatt tgtctcaaga tctagttacg ccaagcttaa aaaagcaccg actcggtgcc
     1321 actttttcaa gttgataacg gactagcctt attttaactt gctatttcta gctctaaaac
     1381 agagacgtcc gtctccggtg tttcgtcctt tccacaagat atataaagcc aagaaatcga
     1441 aatactttca agttacggta agcatatgat agtccatttt aaaacataat tttaaaactg
     1501 caaactaccc aagaaattat tactttctac gtcacgtatt ttgtactaat atctttgtgt
     1561 ttacagtcaa attaattcta attatctctc taacagcctt gtatcgtata tgcaaatatg
     1621 aaggaatcat gggaaatagg ccctctctgg gtcccctcgg ggttgggagg tgggtctgaa
     1681 acgataatgg tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta
     1741 ttcttaaacc taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat
     1801 ttgttatgtt aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt
     1861 cattcttttc ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat
     1921 tgttaatttc tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt
     1981 tattactcca actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac
     2041 cccaaatccc caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct
     2101 ggagctgctt gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc
     2161 tcagcaaatt gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca
     2221 gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag
     2281 ctcctattcc cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg
     2341 ctactcctaa tggttcaatt tttactactt tatatttata taattcactt ctccaattgt
     2401 ccctcatatc tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta
     2461 cttttgtttt gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc
     2521 ctttgatgca cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat
     2581 cctgtctgaa gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct
     2641 aacaggccag gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt
     2701 aatttatatt ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt
     2761 ctcccccgct taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc
     2821 aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca
     2881 agccgagtcc tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc
     2941 gggcgccact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     3001 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     3061 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     3121 agagagaccc gtttatgtat cgagctaggc acttaaatac aatatctctg caatgcggca
     3181 attcagtggt tcgtccaatc catgtcagac ccgtctgttg ccttcctaat aaggcacgat
     3241 cgtaccacct tacttccacc aatcggcatg cacggtgctt tttctctcct tgtaaggcat
     3301 gttgctaact catcgttacc atgttgcaag actacaagag tattgcataa gactacatta
     3361 agcttgcagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     3421 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3481 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3541 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3601 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3661 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3721 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3781 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3841 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3901 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3961 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4021 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4081 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4141 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4201 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4261 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4321 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4381 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4441 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4501 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4561 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4621 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4681 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4741 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4801 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4861 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4921 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4981 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5041 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5101 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5161 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5221 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5281 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5341 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5401 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5461 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5521 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     5581 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5641 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5701 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5761 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5821 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5881 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5941 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6001 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     6061 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     6121 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     6181 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac gtccctaggc
     6241 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     6301 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     6361 ttaggggcgg gatgggcgga gttaggggcg ggatagctag agccagacat gataagatac
     6421 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     6481 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttcctctc
     6541 actctctgat attcatttct ttgcaagtta taaatactga ataataagat gacatgaact
     6601 actatacgta ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt
     6661 accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg
     6721 aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa
     6781 ccagagagac ccagtacaag caaaaagcag atcttgtctt cgttgggagt gaattagccc
     6841 ttccagtccc cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat
     6901 tggtcttaaa gtcgacgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga
     6961 aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct
     7021 ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca
     7081 caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg
     7141 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     7201 gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc
     7261 caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg
     7321 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa
     7381 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
     7441 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
     7501 tgattgtcga cttaacgcgt gaattcttac ttgtacagct cgtccatgcc gtacaggaac
     7561 aggtggtggc ggccctcgga gcgctcgtac tgttccacga tggtgtagtc ctcgttgtgg
     7621 gaggtgatgt ccagcttggt gtccacgtag tagtagccgg gcagttgcac gggcttcttg
     7681 gccatgtaga tggtcttgaa ctccaccagg tagtggccgc cgtccttcag cttcagggcc
     7741 tggtggatct cgcccttcag cacgccgtcg cgggggtaca ggcgctcggt ggaggcctcc
     7801 cagcccatgg tcttcttctg cattacgggg ccgtcggggg ggaagttggt gccgcgcatc
     7861 ttcaccttgt agatcagcgt gccgtcctgc agggaggagt cctgggtcac ggtcaccaga
     7921 ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa ggacagcttc
     7981 ttgtaatcgg ggatgtcggc ggggtgcttc acgtacgcct tggagccgta catgaactgg
     8041 ggggacagga tgtcccaggc gaagggcagg gggccgccct tggtcacctt cagcttggcg
     8101 gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg
     8161 ttcatggagc cctccatgcg caccttgaag cgcatgaact ctttgatgac ctcctcgccc
     8221 ttgctcacca tggtggcggg atcctctaga gtcgacctgc agcccaagct tcgtctaaca
     8281 aaaaagccaa aaacggccag aatttagcgg acaatttact agtctaacac tgaaaattac
     8341 atattgaccc aaatgattac atttcaaaag gtgcctaaaa aacttcacaa aacacactcg
     8401 ccaaccccga gcgcatagtt caaaaccgga g
//
