LOCUS       sequence-462291-       11490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the gRNAs for the human FOXA1/A2/A3-CRISPRi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11490)
  AUTHORS   Matsui S, Granitto M, Buckley M, Ludwig K, Koigi S, Shiley J, 
            Zacharias WJ, Mayhew CN, Lim HW, Iwafuchi M
  TITLE     Pioneer and PRDM transcription factors coordinate bivalent 
            epigenetic states to safeguard cell fate.
  JOURNAL   Mol Cell. 2024 Jan 4:S1097-2765(23)01026-2. doi: 
            10.1016/j.molcel.2023.12.007.
  PUBMED    38211589
REFERENCE   2  (bases 1 to 11490)
  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: "Mol Cell. 
            2024 Jan 4:S1097-2765(23)01026-2. doi: 
            10.1016/j.molcel.2023.12.007."
COMMENT     Sequence Label: pLV-hUbC-EGFP-sgRNA.FOXA1/A2/A3_CRISPRi
FEATURES             Location/Qualifiers
     source          1..11490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2602..2635)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2640..2953
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2872..2892
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        2975..3050
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(3080..3155)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(3176..3424)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(3234..3253)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(3404..3424)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        3454..3668
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     3589..3608
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        3697..3772
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(3802..3877)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4173..5384
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5221..5240
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      5432..5441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5438..5440,5441..5443,5444..6157)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5438..5440,5441..5443,5444..6154)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5483..5504)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5744..5763)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6091..6112
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6182..6770
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6235..6255)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6653..6664)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6773..6789)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6774..6790)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7295..7475
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7501..7518)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7507..7731
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7777..8205
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7864..7883)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8074..8095
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8214..8234)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8219..8548
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8399..8534
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8461..8480
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8596..8643
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8662..9036
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    9166..9287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9203..9222)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9257..9276
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9336..9352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9336..9352)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9349..9371)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9360..9376
                     /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(9384..9390,9391..9408,9409..9414))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9429..9450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9567..9584)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9738..10326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9818..9837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10497..11288,11289..11357))
                     /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     11120..11139
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11358..11462)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aataacttcg tatagcatac attatacgaa gttatgatag
     2641 atccgacgcc gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc
     2701 tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct
     2761 taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat
     2821 aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa acagcacaaa
     2881 aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga
     2941 gaaaagcctt gtttgagcgc tctggatctc tcaggtttta gagctagaaa tagcaagtta
     3001 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgta
     3061 cgctcagcgg agaaaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta
     3121 gccttatttt aacttgctat ttctagctct aaaaccggga agtgagcggg ctgccggtgt
     3181 ttcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa gttacggtaa
     3241 gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca agaaattatt
     3301 actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa ttaattccaa
     3361 ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg ggaaataggc
     3421 cctcttcctg cccgaccttg gatcgatccg cctgaacgct gacgtcatca acccgctcca
     3481 aggaatcgcg ggcccagtgt cactaggcgg gaacacccag cgcgcgtgcg ccctggcagg
     3541 aagatggctg tgagggacag gggagtggcg ccctgcaata tttgcatgtc gctatgtgtt
     3601 ctgggaaatc accataaacg tgaaatgtct ttggatttgg gaatcttata agttctgtat
     3661 gagaccactc tttcccaagt agctgctcca gtcggagttt tagagctaga aatagcaagt
     3721 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttg
     3781 gtcatcagtc cgagaaaaaa agcaccgact cggtgccact ttttcaagtt gataacggac
     3841 tagccttatt ttaacttgct atttctagct ctaaaacgag gtaagcgctc gtctttcgag
     3901 gtacccaagc ggcgcacaag ctatataaac ctgaaggaag tctcaacttt acacttaggt
     3961 caagttgctt atcgtactag agcttcagca ggaaatttaa ctaaaatcta atttaaccag
     4021 catagcaaat atcatttatt cccaaaatgc taaagtttga gataaacgga cttgatttcc
     4081 ggctgttttg acactatcca gaatgccttg cagatgggtg gggcatgcta aatactgcag
     4141 acaattaatt aacccgtgtc ggctccagat ctggcctccg cgccgggttt tggcgcctcc
     4201 cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg
     4261 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
     4321 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
     4381 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
     4441 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
     4501 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
     4561 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
     4621 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
     4681 aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct
     4741 tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg
     4801 tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg cgggaaagct cttattcggg
     4861 tgagatgggc tggggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     4921 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     4981 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     5041 atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
     5101 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     5161 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     5221 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     5281 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagactagta
     5341 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgaagct tgggctgcag
     5401 gtcgactcta gaggatcccc gggtaccggt cgccaccatg gtgagcaagg gcgaggagct
     5461 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     5521 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     5581 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     5641 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     5701 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     5761 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     5821 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     5881 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     5941 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     6001 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     6061 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     6121 cgggatcact ctcggcatgg acgagctgta caagtaagaa ttcgatatca agcttatcga
     6181 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     6241 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6301 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6361 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6421 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6481 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6541 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     6601 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6661 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6721 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     6781 tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     6841 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     6901 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6961 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     7021 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg
     7081 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa
     7141 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg
     7201 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca
     7261 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg
     7321 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     7381 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     7441 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
     7501 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     7561 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     7621 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     7681 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     7741 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     7801 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     7861 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     7921 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     7981 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     8041 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     8101 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     8161 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     8221 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     8281 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     8341 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     8401 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     8461 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     8521 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     8581 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     8641 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     8701 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     8761 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     8821 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     8881 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     8941 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     9001 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     9061 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     9121 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     9181 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     9241 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     9301 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     9361 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     9421 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     9481 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     9541 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     9601 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     9661 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     9721 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     9781 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     9841 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     9901 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     9961 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    10021 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    10081 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    10141 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    10201 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    10261 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    10321 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    10381 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    10441 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    10501 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    10561 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    10621 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    10681 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    10741 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    10801 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    10861 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    10921 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    10981 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    11041 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    11101 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    11161 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    11221 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    11281 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    11341 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    11401 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    11461 cgcacatttc cccgaaaagt gccacctgac
//
