LOCUS       sequence-437718-       10271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To inducibly knockdown CCND1 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10271)
  AUTHORS   Magrath JW, Sampath SS, Flinchum DA, Hartono AB, Goldberg IN, 
            Boehling JR, Savkovic SD, Lee SB
  TITLE     Comprehensive Transcriptomic Analysis of EWSR1::WT1 Targets 
            Identifies CDK4/6 Inhibitors as an Effective Therapy for 
            Desmoplastic Small Round Cell Tumors.
  JOURNAL   Cancer Res. 2024 May 2;84(9):1426-1442. doi: 
            10.1158/0008-5472.CAN-23-3334.
  PUBMED    38588409
REFERENCE   2  (bases 1 to 10271)
  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: "10.1158/0008-5472";
            journalName: "Cancer Res"; date: "2024-05-2- 2"; volume: "84"; 
            issue: "9"; pages: "1426-1442"
COMMENT     Sequence Label: LT3GEPIR-CCND1shRNA3
FEATURES             Location/Qualifiers
     source          1..10271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..31
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    148..169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(184..189,190..207,208..214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    222..238
                     /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     227..249
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     246..262
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     246..262
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     281..301
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        283..301
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        329..555
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             556..736
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    783..908
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1401..1634
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1819..1863
                     /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    2130..2247
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2263..2641
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2271..2289
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2307..2325
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2343..2361
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2379..2397
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2415..2433
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2451..2469
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2487..2505
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2550..2574
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2665..2667,2668..2670,2671..3384)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2710..2731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2971..2990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3318..3339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           3577..3591
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        3770..4269
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3805..3828)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4202..4221
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4290..4889
                     /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(4290..4309)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4786..4806
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4950..5517
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4967..5519
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5111..5128)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5338..5357
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5531..6148,6149..6154,6155..6190,6191..6193,6194..6229,
                     6230..6232,6233..6268,6269..6277)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    6323..6911
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6376..6396)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6794..6805)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7118..7351
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7423..7544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7460..7479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7514..7533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7584..7719
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7739..7758)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7740..7758)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7768..7785)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7768..7784)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7777..7799)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7926..8381
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8013..8032)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8223..8244
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8407..8511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8512..8580,8581..9372)
                     /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(8730..8749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9543..10131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10032..10051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
       61 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      121 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      181 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      241 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
      301 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
      361 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
      421 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
      481 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
      541 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      601 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      661 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      721 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
      781 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
      841 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
      901 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
      961 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     1021 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     1081 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     1141 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     1201 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     1261 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     1321 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     1381 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     1441 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     1501 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     1561 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     1621 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     1681 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     1741 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     1801 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     1861 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     1921 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     1981 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
     2041 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
     2101 gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac
     2161 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     2221 aaacaaatta caaaaattca aaattttatc gattctagat tcgagtttac tccctatcag
     2281 tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta tgcagacttt
     2341 actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg atagagaacg
     2401 tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac tccctatcag
     2461 tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta taagctttag
     2521 gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg
     2581 agcaattcca caacactttt gtcttatacc aactttccgt accacttcct accctcgtaa
     2641 agtcgagctt gcgttggatc caccatggtg agcaagggcg aggagctgtt caccggggtg
     2701 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
     2761 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2821 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2881 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2941 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3001 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3061 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3121 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     3181 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3241 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3301 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3361 ggcatggacg agctgtacaa gtaatgacgt acggcggccg cctcgactag ggataacagg
     3421 gtaattgttt gaatgaggct tcagtacttt acagaatcgt tgcctgcaca tcttggaaac
     3481 acttgctggg attacttcga cttcttaacc caacagaagg ctcgagaagg tatattgctg
     3541 ttgacagtga gcgaattgga atagcttctg gaattagtga agccacagat gtaattccag
     3601 aagctattcc aatgtgccta ctgcctcgga cttcaagggg tctagaattc gagcaattat
     3661 cttgtttact aaaactgaat accttgctat ctctttgata catttttaca aagctgaatt
     3721 aaaatggtat aaattaaatc acttttttca attgacgcgt aattctaccg ggtaggggag
     3781 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     3841 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     3901 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     3961 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     4021 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     4081 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     4141 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     4201 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     4261 ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg
     4321 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     4381 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     4441 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     4501 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     4561 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     4621 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     4681 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     4741 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     4801 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     4861 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
     4921 aaaggagcgc acgaccccat gcatccaatt ccgccccccc cccctaacgt tactggccga
     4981 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     5041 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     5101 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     5161 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     5221 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     5281 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     5341 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     5401 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     5461 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     5521 ggccacaacc atgagtagac tggacaagag caaagtcata aacggagctc tggaattact
     5581 caatggtgtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc tgggagttga
     5641 gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg ccctgccaat
     5701 cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt catggcaaga
     5761 ctttctgcgg aacaacgcca agtcataccg ctgtgctctc ctctcacatc gcgacggggc
     5821 taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg aaaatcagct
     5881 cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc tgtccgccgt
     5941 gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag caaaagagga
     6001 aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa ttgagctgtt
     6061 cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca tatgtggcct
     6121 ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg attttgactt
     6181 agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc ctgctgacgc
     6241 tcttgacgat tttgaccttg acatgctccc cgggtaatcc ggactcgacc tgcagccaag
     6301 cttatcgaaa cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac
     6361 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     6421 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     6481 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     6541 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     6601 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     6661 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     6721 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     6781 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     6841 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     6901 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca
     6961 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     7021 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     7081 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     7141 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     7201 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     7261 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     7321 cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag
     7381 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     7441 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     7501 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     7561 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     7621 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     7681 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     7741 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     7801 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     7861 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     7921 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     7981 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     8041 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     8101 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     8161 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     8221 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     8281 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     8341 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     8401 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     8461 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     8521 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     8581 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     8641 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     8701 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     8761 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8821 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8881 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8941 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     9001 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     9061 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     9121 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     9181 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     9241 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     9301 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     9361 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     9421 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     9481 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     9541 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     9601 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     9661 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     9721 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     9781 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     9841 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     9901 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     9961 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    10021 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    10081 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
    10141 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
    10201 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
    10261 cgccgcagcc g
//
