LOCUS       sequence-428730-       13667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FFSS-Cyclin D1 (HP) in mammalian cells from an inducible 
            lentiviral vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13667)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 13667)
  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 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: pTRIPZ-FFSS-Cyclin D1 (HP)
FEATURES             Location/Qualifiers
     source          1..13667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /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"
     regulatory      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2722..2740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2793..2811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2864..2882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2900..2918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2930..2968
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2965..2989
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3001..3024
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3040..3063
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3079..3102
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3139..3162
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     regulatory      3779..3788
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     ncRNA           4123..4137
                     /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"
     ncRNA           4165..4291
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        complement(4386..4404)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4417..5628
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5465..5484
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(5641..5674)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     CDS             5681..6388
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     protein_bind    complement(6389..6422)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     misc_feature    6445..7017
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6612..6629)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6839..6858
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7020..7622
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7519..7539
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7638..8226
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7691..7711)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8109..8120)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8433..8666
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8767..8784)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8773..8997
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9043..9471
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9130..9149)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9340..9361
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9480..9500)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9485..9814
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9665..9800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9727..9746
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    11022..11143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11192..11208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11192..11208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11205..11227)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11216..11232
                     /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(11240..11246,11247..11264,11265..11270))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11285..11306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11423..11440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11594..12182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11674..11693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12353..13144,13145..13213))
                     /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     12976..12995
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13214..13318)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13366..13500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc
     2761 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
     2821 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga
     2881 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt
     2941 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg caccggtatg
     3001 gactacaagg acgacgatga caaggtacca ggatctgggg attataaaga tgatgatgat
     3061 aaagggtcgg ccgccagctg gagccaccct cagttcgaga agggaggagg aagcggcgga
     3121 ggcagcggag gaggaagctg gagccacccg cagttcgaga aaggagctgg atccagcagc
     3181 gaacaccagc tcctgtgctg cgaagtggaa accatccgcc gcgcgtaccc cgatgccaac
     3241 ctcctcaacg accgggtgct gcgggccatg ctgaaggcgg aggagacctg cgcgccctcg
     3301 gtgtcctact tcaaatgtgt gcagaaggag gtcctgccgt ccgcgcggaa gatcgccgcc
     3361 accgcgatgc tggaggtctg cgaggaacag aagtgcgagg aggaggtctt cccgctggcc
     3421 atgaactacc tggaccgctt cctgtcgctg gagcccgtga aaaagagccg cctgcagctg
     3481 ctgggggcca cttgcatgtt cgtggcctct aagatgaagg agaccatccc cctgacggcc
     3541 gagaagctgt gcatctacac cgacaactcc atccggcccg aggagctgct gcaaatggag
     3601 ctgctcctgg tgaacaagct caagtggaac ctggccgcaa tgaccccgca cgatttcatt
     3661 gaacacttcc tctccaaaat gccagaggcg gaggagaaca aacagatcat ccgcaaacac
     3721 gcgcagacct tcgttgccct ctgtgccaca gatgtgaagt tcatttccaa tccgccctcc
     3781 atggtggcag cggggagcgt ggtggccgca gtgcaaggcc tgaacctgag gagccccaac
     3841 aacttcctgt cctactaccg cctcacacgc ttcctctcca gagtgatcaa gtgtgaccca
     3901 gactgcctcc gggcctgcca ggagcagatc gaagccctgc tggagtcaag cctgcgccag
     3961 gcccagcaga acatggaccc caaggccgcc gaggaggagg aagaggagga ggaggaggtg
     4021 gacctggctt gcacacccac cgacgtgcgg gacgtggaca tctgactcga gaaggtatat
     4081 tgctgttgac agtgagcgat ctcgcttggg cgagagtaag tagtgaagcc acagatgtac
     4141 ttactctcgc ccaagcgaga gtgcctactg cctcggaatt caaggggcta ctttaggagc
     4201 aattatcttg tttactaaaa ctgaatacct tgctatctct ttgatacatt tttacaaagc
     4261 tgaattaaaa tggtataaat taaatcactt ttttcaattg gaagactaat gcggccggcc
     4321 attactccgt ctcgtgtctt gttgcatatg tctgctggtt tgtttgatgt tgtttgcggg
     4381 cgggccctat agtgagtcgt attacctagg acgcgtggcc tccgcgccgg gttttggcgc
     4441 ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg
     4501 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     4561 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     4621 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     4681 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     4741 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     4801 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     4861 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     4921 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     4981 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     5041 atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt
     5101 cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     5161 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     5221 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     5281 tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg
     5341 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     5401 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     5461 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     5521 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     5581 agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacgc tagcggatcc
     5641 ataacttcgt atagtataca ttatacgaag ttatgccacc atgtctaggc tggacaagag
     5701 caaagtcata aacggagctc tggaattact caatggtgtc ggtatcgaag gcctgacgac
     5761 aaggaaactc gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa
     5821 caagcgggcc ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt
     5881 ctgccccctg gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcataccg
     5941 ctgtgctctc ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga
     6001 gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct
     6061 ggagaacgca ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga
     6121 ggaacaggag catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc
     6181 cccacttctg agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct
     6241 tttcggcctg gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg
     6301 gccgaccgac gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga
     6361 ttttgacctt gacatgctcc ccgggtaaat aacttcgtat agtatacatt atacgaagtt
     6421 atggatccgc ggccgcaaat tccgcccctc tccctccccc ccccctaacg ttactggccg
     6481 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     6541 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     6601 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     6661 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     6721 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     6781 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     6841 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     6901 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     6961 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gataatacca
     7021 tggccaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     7081 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     7141 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     7201 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     7261 agagcgtcga agcgggggcg gtgttcgccg agatcggctc gcgcatggcc gagttgagcg
     7321 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     7381 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     7441 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgctggggtg cccgccttcc
     7501 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     7561 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     7621 gagttcgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     7681 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     7741 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     7801 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     7861 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     7921 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     7981 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     8041 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     8101 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     8161 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     8221 ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag
     8281 caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt
     8341 gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga
     8401 tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag
     8461 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     8521 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     8581 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     8641 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     8701 ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgtt taaacccgct
     8761 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     8821 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     8881 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     8941 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     9001 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     9061 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     9121 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     9181 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     9241 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     9301 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     9361 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     9421 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     9481 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     9541 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     9601 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     9661 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     9721 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     9781 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     9841 tttcggatct gatcagcacg tgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     9901 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     9961 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
    10021 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
    10081 gattccggaa gtgcttgaca ttggggaatt aattcagcga gagcctgacc tattgcatct
    10141 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
    10201 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
    10261 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
    10321 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
    10381 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
    10441 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
    10501 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
    10561 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
    10621 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
    10681 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
    10741 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
    10801 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
    10861 gccccagcac tcgtccgagg gcaaaggaat agcacgtgct acgagatttc gattccaccg
    10921 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
    10981 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
    11041 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    11101 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
    11161 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
    11221 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
    11281 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
    11341 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
    11401 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    11461 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    11521 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    11581 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    11641 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    11701 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    11761 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    11821 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    11881 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    11941 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    12001 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    12061 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    12121 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    12181 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    12241 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    12301 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    12361 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    12421 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    12481 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    12541 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    12601 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    12661 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    12721 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    12781 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    12841 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    12901 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    12961 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    13021 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    13081 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    13141 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    13201 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    13261 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    13321 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt gtttattgca
    13381 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    13441 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
    13501 aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata ccctattacc
    13561 actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga attattttca
    13621 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
