LOCUS       sequence-271751-        8630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of gRNAs targeting intron 13 of murine Bcan 
            and intron 10 of murine Ntrk1. Also constitutively expresses 
            Puromycin linked to TagBFP. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8630)
  AUTHORS   Oldrini B, Curiel-Garcia A, Marques C, Matia V, Uluckan O, 
            Grana-Castro O, Torres-Ruiz R, Rodriguez-Perales S, Huse JT, 
            Squatrito M
  TITLE     Somatic genome editing with the RCAS-TVA-CRISPR-Cas9 system for 
            precision tumor modeling.
  JOURNAL   Nat Commun. 2018 Apr 13;9(1):1466. doi: 10.1038/s41467-018-03731-w.
  PUBMED    29654229
REFERENCE   2  (bases 1 to 8630)
  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: "Nat 
            Commun."; date: "2018-04-13"; volume: "9(1):1466. doi"; pages: " 
            10.1038/s41467-018-03731-w"
COMMENT     Sequence Label: pKLV-gRNA-Bcan-Ntrk1_4
FEATURES             Location/Qualifiers
     source          1..8630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        715..1018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1020..1218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1173..1193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1236..1416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1463..1588
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2085..2318
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2503..2547
                     /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    2845..2962
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3045..3293
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3045..3065
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3216..3235
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3315..3390
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3487..3800
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     3719..3739
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        3824..3899
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3922..4421
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3957..3980)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4354..4373
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4461..5057
                     /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(4461..4480)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4957..4977
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5076..5129
                     /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"
     CDS             5130..5831
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(6186..6203)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6192..6416
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6442..6462)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6442..6460)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6481..6497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6481..6497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6494..6516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6505..6521
                     /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(6529..6535,6536..6553,6554..6559))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6574..6595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6712..6729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6883..7471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6963..6982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7642..8433,8434..8502))
                     /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     8265..8284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8503..8607)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgactagtta
      661 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      721 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      781 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      841 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      901 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      961 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     1021 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1081 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1141 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1201 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     1261 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     1321 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     1381 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     1441 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     1501 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     1561 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1621 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1681 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1741 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1801 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1861 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1921 gcaagcggcc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     1981 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     2041 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     2101 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     2161 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     2221 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     2281 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     2341 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     2401 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     2461 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     2521 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     2581 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     2641 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     2701 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     2761 gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc aattctgcag
     2821 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
     2881 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
     2941 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggtt
     3001 agtaccgggc cctacgcgtt actcgagcca aggtcgggca ggaagagggc ctatttccca
     3061 tgattccttc atatttgcat atacgataca aggctgttag agagataatt agaattaatt
     3121 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     3181 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     3241 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgaaccac
     3301 acttgggctc ctaagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     3361 caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga aatagcaagt
     3421 taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa tggctctaga
     3481 tctagagatc cgacgccgcc atctctaggc ccgcgccggc cccctcgcac agacttgtgg
     3541 gagaagctcg gctactcccc tgccccggtt aatttgcata taatatttcc tagtaactat
     3601 agaggcttaa tgtgcgataa aagacagata atctgttctt tttaatacta gctacatttt
     3661 acatgatagg cttggatttc tataagagat acaaatacta aattattatt ttaaaaaaca
     3721 gcacaaaagg aaactcaccc taactgtaaa gtaattgtgt gttttgagac tataaatatg
     3781 catgcgagaa aagccttgtt tggccggcta gctgagattg ctagttttag agctagaaat
     3841 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     3901 ttttttggat ccaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     3961 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     4021 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     4081 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     4141 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     4201 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     4261 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     4321 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     4381 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcatc catctagatc
     4441 tcgagcagct gaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     4501 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     4561 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     4621 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     4681 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     4741 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     4801 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     4861 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     4921 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     4981 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5041 cccgcaagcc cggtgccggc ggcgggtccg gaggagaggg cagaggaagt ctcctaacat
     5101 gcggtgacgt ggaggagaat cctggcccaa tgagcgagct gattaaggag aacatgcaca
     5161 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
     5221 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
     5281 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
     5341 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
     5401 agagagtcac cacatacgag gacgggggcg tgctgaccgc tacccaggac accagcctcc
     5461 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
     5521 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg
     5581 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     5641 tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     5701 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     5761 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     5821 agcttaattg agcggccgct aggtaccttt aagaccaatg acttacaagg cagctgtaga
     5881 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaaagaag
     5941 tcaagatctg ctttttgcct gtactgggtc tctctggtta gaccagagtc tctctggtta
     6001 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6061 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6121 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagttt aaacccgctg
     6181 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     6241 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     6301 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     6361 gggggaggat tgggaagtca atagcaggca tgctggggat gcggtgggct ctatgggcgg
     6421 ccgttaatta agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     6481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6541 cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7321 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
