LOCUS       sequence-373040-       10966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone to clone single or  paired sgRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10966)
  AUTHORS   Pulido-Quetglas C, Aparicio-Prat E, Arnan C, Polidori T, Hermoso T, 
            Palumbo E, Ponomarenko J, Guigo R, Johnson R
  TITLE     Scalable Design of Paired CRISPR Guide RNAs for Genomic Deletion.
  JOURNAL   PLoS Comput Biol. 2017 Mar 2;13(3):e1005341. doi: 
            10.1371/journal.pcbi.1005341. eCollection 2017 Mar.
  PUBMED    28253259
REFERENCE   2  (bases 1 to 10966)
  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: "PLoS Comput
            Biol."; date: "2017-03-2"; volume: "13(3):e1005341. doi"; pages: " 
            10.1371/journal.pcbi.1005341. eCollection 2017 Mar"
COMMENT     Sequence Label: pDECKO_mCherry
FEATURES             Location/Qualifiers
     source          1..10966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    39..60
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(75..80,81..98,99..105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    113..129
                     /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     118..140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     137..153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     137..153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     172..192
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        174..192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        220..446
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             447..627
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    674..799
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1292..1525
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1710..1754
                     /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        1932..2180
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1932..1952
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2103..2122
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4062..4137
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    4193..4310
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4449..5627
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4680..5618
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5575..5595
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5640..6236
                     /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(5640..5659)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6136..6156
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             6246..6299
                     /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      6306..6315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6312..7022
                     /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(6457..6475)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6725..6744
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    7038..7626
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7091..7111)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7509..7520)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7698..7931
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8009..8130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8046..8065)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8100..8119
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8170..8305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8325..8344)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8326..8344)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8354..8371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8354..8370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8363..8385)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8512..8967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8599..8618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8809..8830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8993..9097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9098..9166,9167..9958)
                     /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(9316..9335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10129..10717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10618..10637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10871..10888
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
       61 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      121 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      181 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
      241 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
      301 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
      361 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
      421 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
      481 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
      541 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
      601 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
      661 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
      721 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
      781 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
      841 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
      901 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
      961 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1021 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1081 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1141 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1201 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1261 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1321 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1381 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1441 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1501 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     1561 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     1621 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1681 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     1741 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     1801 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     1861 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     1921 ggggacccag agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     1981 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata
     2041 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     2101 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     2161 tgtggaaagg acgaaacacc ggagacggtt gtaaatgagc acacaaaata cacatgctaa
     2221 aatattatat tctatgacct ttataaaatc aaccaaaatc ttctttttaa taactttagt
     2281 atcaataatt agaattttta tgttcctttt tgcaaacttt taataaaaat gagcaaaata
     2341 aaaaaacgct agttttagta actcgcgttg ttttcttcac ctttaataat agctactcca
     2401 ccacttgttc ctaagcggtc agctcctgct tcaatcattt tttgagcatc ttcaaatgtt
     2461 ctaactccac cagctgcttt aactaaagca ttgtctttaa caactgactt cattagttta
     2521 acatcttcaa atgttgcacc tgattttgaa aatcctgttg atgttttaac aaattctaat
     2581 ccagcttcaa cagctatttc acaagctttc atgatttctt cttttgttaa taaacaattt
     2641 tccataatac atttaacaac atgtgatcca gctgcttttt ttacagcttt catgtcttct
     2701 aaaactaatt cataattttt gtcttttaat gcaccaatat ttaataccat atcaatttct
     2761 gttgcaccat ctttaattgc ttcagaaact tcgaatgctt ttgtagctgt tgtgcatgca
     2821 cctagaggaa aacctacaac atttgttatt cctacatttg tgccttttaa taattcttta
     2881 caatagcttg ttcaatatga attaacacaa actgttgcaa aatcaaattc aattgcttca
     2941 tcacataatt gtttaatttc agctttcgta gcatcttgtt ttaataatgt gtgatctata
     3001 tatttgttta gtttcatttt ttctcctata tattcatttt taattttaat tctttaataa
     3061 tttcgtctac tttaacttta gcgttttgaa cagattcacc aacacctata aaataaattt
     3121 ttagtttagg ttcagttcca cttgggcgaa cagcaaatca tgacttatct tctaaataaa
     3181 attttagtaa gtcttgtcct ggcatattat acattccatc gatgtagtct tcaacattaa
     3241 caactttaag tccagcaatt tgagttaagg gtgttgctct caatgatttc attaatggtt
     3301 caatttttaa tttcttttct tctggtttaa aattcaagtt taaagtgaaa gtgtaatatg
     3361 cacccatttc tttaaataaa tcttctaaat agtctactaa tgttttattt tgttttttat
     3421 aaaatcaagc agcctctgct attaatatag aagcttgtat tccatcttta tctctagctg
     3481 agtcatcaat tacatatcca taactttctt cataagcaaa aacaaaattt aatccgttat
     3541 cttcttcttt agcaatttct ctacccattc atttaaatcc agttaaagtt tttacaatat
     3601 taactccata tttttcatga gcgattctat cacccaaatc acttgttaca aaacttgaat
     3661 atagagccgg attttttgga atgctattta agcgttttag atttgataat tttcaatcaa
     3721 ttaaaattgg tcctgtttga tttccatcta atcttacaaa atgaccatca tgttttattg
     3781 ccattccaaa tctgtcagca tctgggtcat tcataataat aatatctgca tcatgtttaa
     3841 taccatattc aagcggtatt tttcatgcag gatcaaattc tggatttgga tttacaacat
     3901 ttttaaatgt ttcatcttca aatgcatgct cttcaacctc aataacgtta tatcctgatt
     3961 cacgtaatat ttttggggta aatttagttc ctgttccatt aactgcgcta aaaataattt
     4021 ttaaatcttt tttagcttct tgctcttttt tgtacgtctc tgttttagag ctagaaatag
     4081 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     4141 tttaagcttg gcgtaactag atcttgagac aaatggcagt attcatccac aattttaaaa
     4201 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     4261 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     4321 acagggacag cagagatcca ctttggcgcc ggctcgaggg ggcccgggtg caaagatgga
     4381 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
     4441 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     4501 agttgggggg aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact
     4561 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     4621 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     4681 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
     4741 cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
     4801 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
     4861 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
     4921 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
     4981 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
     5041 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
     5101 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
     5161 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     5221 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     5281 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     5341 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     5401 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     5461 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     5521 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     5581 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgacgt acggccacca
     5641 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     5701 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     5761 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     5821 tcggcaaggt gtgggtcgcg gacgacggcg ccgccgtggc ggtctggacc acgccggaga
     5881 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5941 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     6001 ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag ggtctgggca
     6061 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     6121 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     6181 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccggat
     6241 cgggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccac
     6301 cggtcgccac catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     6361 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     6421 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     6481 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     6541 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     6601 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     6661 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     6721 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     6781 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     6841 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     6901 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     6961 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     7021 aaacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     7081 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     7141 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     7201 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     7261 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     7321 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     7381 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     7441 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     7501 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     7561 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     7621 ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta gccacttttt
     7681 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     7741 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     7801 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     7861 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     7921 aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc
     7981 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa
     8041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     8101 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta
     8161 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     8221 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     8281 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt
     8341 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     8401 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     8461 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     8521 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     8581 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     8641 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     8701 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     8761 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     8821 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     8881 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     8941 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     9001 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     9061 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     9121 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     9181 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     9241 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     9301 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     9361 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     9421 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     9481 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     9541 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     9601 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     9661 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     9721 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     9781 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     9841 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     9901 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     9961 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
    10021 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
    10081 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
    10141 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    10201 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    10261 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
    10321 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
    10381 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
    10441 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
    10501 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
    10561 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
    10621 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
    10681 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
    10741 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
    10801 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
    10861 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
    10921 tctccccgcg cgttggccga ttcattaatg cagctggcac gacagg
//
