LOCUS       sequence-428727-       10013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Makes CRISPR gRNAs detectable by single-cell RNA-seq. The gRNA is 
            expressed as part of the blasticidin resistance mRNA transcribed by 
            RNA Pol II and in its functional form from the hU6 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10013)
  AUTHORS   Reicher A, Reinis J, Ciobanu M, Ruzicka P, Malik M, Siklos M, 
            Kartysh V, Tomek T, Koren A, Rendeiro AF, Kubicek S
  TITLE     Pooled multicolour tagging for visualizing subcellular protein 
            dynamics.
  JOURNAL   Nat Cell Biol. 2024 May;26(5):745-756. doi: 
            10.1038/s41556-024-01407-w. Epub 2024 Apr 19.
  PUBMED    38641660
REFERENCE   2  (bases 1 to 10013)
  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: 
            "10.1038/s41556-024-01407-w"; journalName: "Nat Cell Biol"; date: 
            "2024-05"; volume: "26"; issue: "5"; pages: "745-756"
COMMENT     Sequence Label: CROPseq-Guide-BSD
FEATURES             Location/Qualifiers
     source          1..10013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             48..92
                     /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    273..390
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        529..1707
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          760..1698
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1655..1675
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1714..1723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1720..2118
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    2134..2722
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2187..2207)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2605..2616)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        2878..3126
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2878..2898
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3049..3068
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        5008..5083
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     LTR             5136..5316
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5394..5515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5431..5450)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5485..5504
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5555..5690
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5710..5729)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5711..5729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5739..5756)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5739..5755)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5748..5770)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5897..6352
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5984..6003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6194..6215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6378..6482
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6483..6551,6552..7343)
                     /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(6701..6720)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7514..8102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8003..8022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8256..8273
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8390..8411
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8426..8431,8432..8449,8450..8456)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8464..8480
                     /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     8469..8491
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8488..8504
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8488..8504
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8523..8543
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8525..8543
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8571..8797
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8798..8978
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9025..9150
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9643..9876
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
       61 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
      121 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
      181 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
      241 atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
      301 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
      361 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
      421 ctttggcgcc ggctcgaggg ggcccgggtg caaagatgga taaagtttta aacagagagg
      481 aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaattgg ctccggtgcc
      541 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
      601 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
      661 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
      721 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc
      781 cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg
      841 ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg
      901 ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
      961 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     1021 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     1081 gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     1141 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     1201 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     1261 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     1321 gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg
     1381 ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc
     1441 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc
     1501 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     1561 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     1621 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     1681 ttttttcttc catttcaggt gtcgtgacgt acggccacca tggccaagcc tttgtctcaa
     1741 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
     1801 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     1861 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     1921 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     1981 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     2041 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     2101 tatgtgtggg agggctaaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     2161 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2221 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2281 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2341 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2401 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2461 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2521 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2581 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2641 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2701 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     2761 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     2821 gacaagattt caccattatc gtttcagacc cacctcccaa ccccgagggg acccagagag
     2881 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     2941 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     3001 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     3061 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     3121 aacaccggag acggttgtaa atgagcacac aaaatacaca tgctaaaata ttatattcta
     3181 tgacctttat aaaatcaacc aaaatcttct ttttaataac tttagtatca ataattagaa
     3241 tttttatgtt cctttttgca aacttttaat aaaaatgagc aaaataaaaa aacgctagtt
     3301 ttagtaactc gcgttgtttt cttcaccttt aataatagct actccaccac ttgttcctaa
     3361 gcggtcagct cctgcttcaa tcattttttg agcatcttca aatgttctaa ctccaccagc
     3421 tgctttaact aaagcattgt ctttaacaac tgacttcatt agtttaacat cttcaaatgt
     3481 tgcacctgat tttgaaaatc ctgttgatgt tttaacaaat tctaatccag cttcaacagc
     3541 tatttcacaa gctttcatga tttcttcttt tgttaataaa caattttcca taatacattt
     3601 aacaacatgt gatccagctg ctttttttac agctttcatg tcttctaaaa ctaattcata
     3661 atttttgtct tttaatgcac caatatttaa taccatatca atttctgttg caccatcttt
     3721 aattgcttca gaaacttcga atgcttttgt agctgttgtg catgcaccta gaggaaaacc
     3781 tacaacattt gttattccta catttgtgcc ttttaataat tctttacaat agcttgttca
     3841 atatgaatta acacaaactg ttgcaaaatc aaattcaatt gcttcatcac ataattgttt
     3901 aatttcagct ttcgtagcat cttgttttaa taatgtgtga tctatatatt tgtttagttt
     3961 cattttttct cctatatatt catttttaat tttaattctt taataatttc gtctacttta
     4021 actttagcgt tttgaacaga ttcaccaaca cctataaaat aaatttttag tttaggttca
     4081 gttccacttg ggcgaacagc aaatcatgac ttatcttcta aataaaattt tagtaagtct
     4141 tgtcctggca tattatacat tccatcgatg tagtcttcaa cattaacaac tttaagtcca
     4201 gcaatttgag ttaagggtgt tgctctcaat gatttcatta atggttcaat ttttaatttc
     4261 ttttcttctg gtttaaaatt caagtttaaa gtgaaagtgt aatatgcacc catttcttta
     4321 aataaatctt ctaaatagtc tactaatgtt ttattttgtt ttttataaaa tcaagcagcc
     4381 tctgctatta atatagaagc ttgtattcca tctttatctc tagctgagtc atcaattaca
     4441 tatccataac tttcttcata agcaaaaaca aaatttaatc cgttatcttc ttctttagca
     4501 atttctctac ccattcattt aaatccagtt aaagttttta caatattaac tccatatttt
     4561 tcatgagcga ttctatcacc caaatcactt gttacaaaac ttgaatatag agccggattt
     4621 tttggaatgc tatttaagcg ttttagattt gataattttc aatcaattaa aattggtcct
     4681 gtttgatttc catctaatct tacaaaatga ccatcatgtt ttattgccat tccaaatctg
     4741 tcagcatctg ggtcattcat aataataata tctgcatcat gtttaatacc atattcaagc
     4801 ggtatttttc atgcaggatc aaattctgga tttggattta caacattttt aaatgtttca
     4861 tcttcaaatg catgctcttc aacctcaata acgttatatc ctgattcacg taatattttt
     4921 ggggtaaatt tagttcctgt tccattaact gcgctaaaaa taatttttaa atctttttta
     4981 gcttcttgct cttttttgta cgtctctgtt ttagagctag aaatagcaag ttaaaataag
     5041 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttta agcttggcgt
     5101 aactagatct tgagacactg ctttttgctt gtactgggtc tctctggtta gaccagatct
     5161 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5221 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5281 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     5341 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     5401 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5461 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5521 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5581 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5641 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     5701 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     5761 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5821 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5881 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5941 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6001 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6061 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6121 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6181 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6241 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6301 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     6361 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6421 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6481 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6541 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6601 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6661 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6721 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6781 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6841 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     6901 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6961 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7021 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7081 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7141 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7201 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7261 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7321 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7381 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7441 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7501 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7561 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7621 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7681 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7741 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7801 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7861 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     7921 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     7981 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8041 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8101 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8161 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     8221 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     8281 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     8341 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     8401 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     8461 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     8521 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
     8581 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
     8641 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
     8701 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
     8761 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
     8821 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     8881 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     8941 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     9001 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     9061 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     9121 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     9181 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     9241 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     9301 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     9361 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     9421 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     9481 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     9541 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     9601 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     9661 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     9721 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     9781 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     9841 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     9901 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     9961 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt aca
//
