LOCUS       sequence-459266-        8889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting Cdkn1a, mCherry, puromycin resistance, Mammalian 
            expression, Mouse targeting, lentiviral, CRISPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8889)
  AUTHORS   Boutelle AM, Mabene AR, Yao D, Xu H, Wang M, Tang YJ, Lopez SS, 
            Sinha S, Demeter J, Cheng R, Benard BA, McCrea EM, Valente LJ, 
            Drainas AP, Fischer M, Majeti R, Petrov DA, Jackson PK, Yang F, 
            Winslow MM, Bassik MC, Attardi LD
  TITLE     Integrative multiomic approaches reveal ZMAT3 and p21 as conserved 
            hubs in the p53 tumor suppression network.
  JOURNAL   Cell Death Differ. 2025 Apr 22. doi: 10.1038/s41418-025-01513-8.
  PUBMED    40263541
REFERENCE   2  (bases 1 to 8889)
  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: "Cell Death 
            Differ. 2025 Apr 22. doi: 10.1038/s41418-025-01513-8."
COMMENT     Sequence Label: pMCB320 sgCdkn1a.1
FEATURES             Location/Qualifiers
     source          1..8889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             750..930
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    977..1102
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1599..1832
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2017..2061
                     /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    2359..2476
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2729..2762)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2766..2786
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     promoter        3031..3242
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          3243..4181
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4138..4158
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4202..4798
                     /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(4202..4221)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4698..4718
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4808..4861
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      4868..4877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4874..5584
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5019..5037)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5287..5306
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(5604..5637)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5693..6281
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5746..5766)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6164..6175)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6284..6300)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6285..6301)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6810..6990
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7052..7640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7132..7151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7811..8602,8603..8671))
                     /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     8434..8453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8672..8776)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8851..8870)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
       61 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
      121 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      781 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      841 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      901 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      961 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     1021 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     1081 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     1141 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1201 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1261 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1321 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1381 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1441 ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1501 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1561 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1621 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1681 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1741 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1801 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1861 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1921 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1981 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     2041 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     2101 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     2161 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     2221 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     2281 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     2341 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     2401 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     2461 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
     2521 gggcccgctc tagagatccg acgcgccatc tctaggcccg cgccggcccc ctcgcacgga
     2581 cttgtgggag aagctcggct actcccctgc cccggttaat ttgcatataa tatttcctag
     2641 taactataga ggcttaatgt gcgataaaag acagataatc tgttcttttt aatactagct
     2701 acattttaca tgataggctt ggatttctat aacttcgtat agcatacatt atacgaagtt
     2761 ataaacagca caaaaggaaa ctcaccctaa ctgtaaagta attgtgtgtt ttgagactat
     2821 aagtatccct tggagaacca ccttgttggg aacaggtcgg acatcaccgt ttaagagcta
     2881 agctggaaac agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg
     2941 caccgagtcg gtgctttttt tctcgagtac taggatccat taggcggccg cgtggataac
     3001 cgtattaccg ccatgcatgt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     3061 agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
     3121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     3181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     3241 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
     3301 tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg
     3361 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag
     3421 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct
     3481 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc
     3541 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg
     3601 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc
     3661 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg
     3721 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc
     3781 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct
     3841 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa
     3901 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca
     3961 ggcacctcga ttagttctcg cgcttttgga gtacgtcgtc tttaggttgg ggggaggggt
     4021 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
     4081 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
     4141 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga gctagcccac
     4201 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     4261 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     4321 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     4381 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     4441 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     4501 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     4561 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     4621 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     4681 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     4741 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgg
     4801 atcgggagag ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc
     4861 accggtcgcc accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     4921 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     4981 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     5041 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     5101 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     5161 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     5221 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     5281 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     5341 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     5401 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     5461 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     5521 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     5581 gtaagaattc gtcgagggac ctaataactt cgtatagcat acattatacg aagttataca
     5641 tgtttaaggg ttccggttcc actaggtaca attcgatatc aagcttatcg ataatcaacc
     5701 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     5761 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     5821 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     5881 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     5941 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     6001 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6061 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     6121 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6181 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6241 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     6301 atcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     6361 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     6421 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6481 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
     6541 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
     6601 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
     6661 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
     6721 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
     6781 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
     6841 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6901 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6961 ccttttagtc agtgtggaaa atctctagca gcatgtgagc aaaaggccag caaaaggcca
     7021 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     7081 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     7141 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     7201 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7261 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7321 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7381 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7441 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7501 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7561 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7621 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7681 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7741 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7801 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7861 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7921 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7981 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     8041 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     8101 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     8161 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     8221 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     8281 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8341 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8401 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8461 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8521 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8581 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8641 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8701 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8761 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8821 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     8881 ccagtatct
//
