LOCUS       sequence-354474-       10224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A version of the CROP-seq plasmid as presented in Datlinger et al. 
            that contains the sgRNA-(F+E)-combined optimized backbone for 
            CRISPRi from Chen et al..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10224)
  AUTHORS   Hill AJ, McFaline-Figueroa JL, Starita LM, Gasperini MJ, Matreyek 
            KA, Packer J, Jackson D, Shendure J, Trapnell C
  TITLE     On the design of CRISPR-based single-cell molecular screens.
  JOURNAL   Nat Methods. 2018 Feb 19. pii: nmeth.4604. doi: 10.1038/nmeth.4604.
  PUBMED    29457792
REFERENCE   2  (bases 1 to 10224)
  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 
            Methods. 2018 Feb 19. pii: nmeth.4604. doi: 10.1038/nmeth.4604."
COMMENT     Sequence Label: CROP-seq-opti
FEATURES             Location/Qualifiers
     source          1..10224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     652..672
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        654..672
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        700..926
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             927..1107
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1154..1279
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1772..2005
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2190..2234
                     /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    2415..2532
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2671..3849
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2902..3840
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3797..3817
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3862..4461
                     /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(3862..3881)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4358..4378
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4477..5065
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4530..4550)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4948..4959)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5221..5469
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5221..5241
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5392..5411
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     LTR             7489..7669
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    7747..7868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7784..7803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7838..7857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7908..8043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8063..8082)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8064..8082)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8092..8109)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8092..8108)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8101..8123)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8250..8705
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8337..8356)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8547..8568
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8731..8835
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8836..8904,8905..9696)
                     /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(9054..9073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(9867..10224,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      661 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
      721 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
      781 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
      841 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
      901 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
      961 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     1021 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     1081 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     1141 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     1201 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     1261 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     1321 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1381 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1441 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1501 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1561 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1621 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1681 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1741 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1801 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1861 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1921 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1981 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     2041 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     2101 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     2161 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     2221 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     2281 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     2341 tgaatagagt taggcaggga tattcaccat tatcgtttca gaatcttgag acaaatggca
     2401 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     2461 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     2521 attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag
     2581 ggggcccggg tgcaaagatg gataaagttt taaacagaga ggaatctttg cagctaatgg
     2641 accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc
     2701 acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgatc cggtgcctag
     2761 agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc
     2821 gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac
     2881 gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt
     2941 acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta cgtgattctt
     3001 gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc
     3061 ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg cgtgcgaatc
     3121 tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat ttaaaatttt
     3181 tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc gggccaagat
     3241 ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg tgcgtcccag
     3301 cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg acgggggtag
     3361 tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc
     3421 tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg gccgcttccc
     3481 ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg ggcgggtgag
     3541 tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg tgactccacg
     3601 gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct
     3661 ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact
     3721 gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt
     3781 ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag
     3841 gtgtcgtgac gtacggccac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3901 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3961 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     4021 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgccgtg
     4081 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     4141 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4201 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac
     4261 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     4321 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4381 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4441 acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt
     4501 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4561 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     4621 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     4681 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     4741 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     4801 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     4861 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     4921 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     4981 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5041 atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg
     5101 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     5161 gaagacaaga tttcaccatt atcgtttcag acccacctcc caaccccgag gggacccaga
     5221 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     5281 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     5341 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     5401 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     5461 cgaaacaccg gagacggttg taaatgagca cacaaaatac acatgctaaa atattatatt
     5521 ctatgacctt tataaaatca accaaaatct tctttttaat aactttagta tcaataatta
     5581 gaatttttat gttccttttt gcaaactttt aataaaaatg agcaaaataa aaaaacgcta
     5641 gttttagtaa ctcgcgttgt tttcttcacc tttaataata gctactccac cacttgttcc
     5701 taagcggtca gctcctgctt caatcatttt ttgagcatct tcaaatgttc taactccacc
     5761 agctgcttta actaaagcat tgtctttaac aactgacttc attagtttaa catcttcaaa
     5821 tgttgcacct gattttgaaa atcctgttga tgttttaaca aattctaatc cagcttcaac
     5881 agctatttca caagctttca tgatttcttc ttttgttaat aaacaatttt ccataataca
     5941 tttaacaaca tgtgatccag ctgctttttt tacagctttc atgtcttcta aaactaattc
     6001 ataatttttg tcttttaatg caccaatatt taataccata tcaatttctg ttgcaccatc
     6061 tttaattgct tcagaaactt cgaatgcttt tgtagctgtt gtgcatgcac ctagaggaaa
     6121 acctacaaca tttgttattc ctacatttgt gccttttaat aattctttac aatagcttgt
     6181 tcaatatgaa ttaacacaaa ctgttgcaaa atcaaattca attgcttcat cacataattg
     6241 tttaatttca gctttcgtag catcttgttt taataatgtg tgatctatat atttgtttag
     6301 tttcattttt tctcctatat attcattttt aattttaatt ctttaataat ttcgtctact
     6361 ttaactttag cgttttgaac agattcacca acacctataa aataaatttt tagtttaggt
     6421 tcagttccac ttgggcgaac agcaaatcat gacttatctt ctaaataaaa ttttagtaag
     6481 tcttgtcctg gcatattata cattccatcg atgtagtctt caacattaac aactttaagt
     6541 ccagcaattt gagttaaggg tgttgctctc aatgatttca ttaatggttc aatttttaat
     6601 ttcttttctt ctggtttaaa attcaagttt aaagtgaaag tgtaatatgc acccatttct
     6661 ttaaataaat cttctaaata gtctactaat gttttatttt gttttttata aaatcaagca
     6721 gcctctgcta ttaatataga agcttgtatt ccatctttat ctctagctga gtcatcaatt
     6781 acatatccat aactttcttc ataagcaaaa acaaaattta atccgttatc ttcttcttta
     6841 gcaatttctc tacccattca tttaaatcca gttaaagttt ttacaatatt aactccatat
     6901 ttttcatgag cgattctatc acccaaatca cttgttacaa aacttgaata tagagccgga
     6961 ttttttggaa tgctatttaa gcgttttaga tttgataatt ttcaatcaat taaaattggt
     7021 cctgtttgat ttccatctaa tcttacaaaa tgaccatcat gttttattgc cattccaaat
     7081 ctgtcagcat ctgggtcatt cataataata atatctgcat catgtttaat accatattca
     7141 agcggtattt ttcatgcagg atcaaattct ggatttggat ttacaacatt tttaaatgtt
     7201 tcatcttcaa atgcatgctc ttcaacctca ataacgttat atcctgattc acgtaatatt
     7261 tttggggtaa atttagttcc tgttccatta actgcgctaa aaataatttt taaatctttt
     7321 ttagcttctt gctctttttt gtacgtctct gtttaagagc tatgctggaa acagcatagc
     7381 aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     7441 ttaagcttgg cgtaactaga tcttgagaca ctgctttttg cttgtactgg gtctctctgg
     7501 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     7561 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     7621 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tacgtatagt
     7681 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     7741 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7801 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7861 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
     7921 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     7981 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     8041 gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
     8101 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     8161 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     8221 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     8281 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     8341 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     8401 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     8461 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     8521 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     8581 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     8641 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     8701 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     8761 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     8821 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     8881 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     8941 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     9001 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     9061 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     9121 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     9181 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     9241 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     9301 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     9361 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     9421 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     9481 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     9541 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     9601 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     9661 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     9721 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     9781 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     9841 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     9901 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     9961 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
    10021 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
    10081 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
    10141 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
    10201 acacagccca gcttggagcg aacg
//
