LOCUS       sequence-175021-        8323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR KO of Trp53.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8323)
  AUTHORS   Wang Q, Zou Y, Nowotschin S, Kim SY, Li QV, Soh CL, Su J, Zhang C, 
            Shu W, Xi Q, Huangfu D, Hadjantonakis AK, Massague J
  TITLE     The p53 Family Coordinates Wnt and Nodal Inputs in Mesendodermal 
            Differentiation of Embryonic Stem Cells.
  JOURNAL   Cell Stem Cell. 2017 Jan 5;20(1):70-86. doi: 
            10.1016/j.stem.2016.10.002. Epub 2016 Nov 23.
  PUBMED    27889317
REFERENCE   2  (bases 1 to 8323)
  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.stem.2016.10.002"; journalName: "Cell Stem Cell"; date: 
            "2017-01-5- 5"; volume: "20"; issue: "1"; pages: "70-86"
COMMENT     Sequence Label: LentiGuidPuro-hTP53_sgRNA-1
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      9..597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     498..517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     751..768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    885..906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(921..926,927..944,945..951)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    959..975
                     /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     964..986
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     983..999
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     983..999
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1018..1038
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1020..1038
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1066..1292
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1293..1473
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1520..1645
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2138..2371
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2556..2600
                     /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        2778..3026
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2778..2798
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2949..2968
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3048..3123
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3179..3296
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3435..4613
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3666..4604
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4561..4581
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4626..5225
                     /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(4626..4645)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5122..5142
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5241..5829
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5294..5314)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5712..5723)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5901..6134
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6212..6333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6249..6268)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6303..6322
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6373..6508
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6528..6547)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6529..6547)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6557..6574)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6557..6573)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6566..6588)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6715..7170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6802..6821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7012..7033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7196..7300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7301..7369,7370..8161)
                     /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(7519..7538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
       61 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      601 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      661 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      721 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      781 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      841 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      901 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      961 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     1021 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     1081 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     1141 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     1201 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     1261 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
     1321 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     1381 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     1441 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     1501 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     1561 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     1621 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     1681 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     1741 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     1801 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1861 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1921 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1981 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     2041 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     2101 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     2161 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     2221 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     2281 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     2341 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     2401 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     2461 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     2521 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     2581 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     2641 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     2701 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     2761 ccccgagggg acccagagag ggcctatttc ccatgattcc ttcatatttg catatacgat
     2821 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
     2881 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     2941 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     3001 atatcttgtg gaaaggacga aacaccggag cgctgctcag atagcgagtt ttagagctag
     3061 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     3121 tgctttttta agcttggcgt aactagatct tgagacaaat ggcagtattc atccacaatt
     3181 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     3241 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     3301 tttattacag ggacagcaga gatccacttt ggcgccggct cgagggggcc cgggtgcaaa
     3361 gatggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga
     3421 aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     3481 ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg
     3541 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     3601 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     3661 cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt
     3721 gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg
     3781 ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt
     3841 gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg
     3901 cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga
     3961 cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt
     4021 cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga
     4081 ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc
     4141 ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg
     4201 cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga
     4261 gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga
     4321 aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt
     4381 ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg
     4441 ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt
     4501 ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc
     4561 tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg
     4621 ccaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     4681 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     4741 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4801 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc cgtggcggtc tggaccacgc
     4861 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4921 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     4981 aggagcccgc gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc
     5041 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     5101 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5161 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5221 cctgaacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     5281 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     5341 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     5401 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     5461 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     5521 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     5581 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     5641 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     5701 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     5761 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     5821 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     5881 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     5941 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6001 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6061 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6121 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
     6181 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     6241 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6301 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     6361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     6421 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     6481 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     6541 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     6601 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     6661 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     6721 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6781 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6841 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6901 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     6961 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     7021 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     7081 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7141 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     7201 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7261 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7321 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7381 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     7441 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     7501 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     7561 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     7621 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     7681 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     7741 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     7801 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7861 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7921 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7981 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     8041 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     8101 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     8161 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     8221 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     8281 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaa
//
