LOCUS       sequence-473629-       14155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  doxycycline inducible expression of E2F3A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14155)
  AUTHORS   Clijsters L, Hoencamp C, Calis JJA, Marzio A, Handgraaf SM, Cuitino 
            MC, Rosenberg BR, Leone G, Pagano M
  TITLE     Cyclin F Controls Cell-Cycle Transcriptional Outputs by Directing 
            the Degradation of the Three Activator E2Fs.
  JOURNAL   Mol Cell. 2019 Jun 20;74(6):1264-1277.e7. doi: 
            10.1016/j.molcel.2019.04.010. Epub 2019 May 23.
  PUBMED    31130363
REFERENCE   2  (bases 1 to 14155)
  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: 
            "10.1016/j.molcel.2019.04.010"; journalName: "Mol Cell"; date: 
            "2019-06-20- 20"; volume: "74"; issue: "6"; pages: "1264-1277.e7"
COMMENT     Sequence Label: pTRIPZ-2xFLAG-2xSTREP-E2F3A
FEATURES             Location/Qualifiers
     source          1..14155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(126..170)
                     /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    355..588
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1085..1210
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1257..1890
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    2058..2192
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2240..2344
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2345..2413,2414..3205)
                     /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(2563..2582)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3376..3964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3865..3884
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4118..4135
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4252..4273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4288..4293,4294..4311,4312..4318)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4326..4342
                     /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     4331..4353
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4350..4366
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4350..4366
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4415..4536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5744..6073)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5758..5893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5812..5831)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6058..6078
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6087..6515)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6197..6218)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6409..6428
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6561..6785
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6774..6791
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6892..7125
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    7332..7920
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7438..7449
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7847..7867
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7936..8538)
                     /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     complement(8019..8039)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8541..9113
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8700..8719)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     8929..8946
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    9136..9169
                     /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             complement(9170..9877)
                     /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    9884..9917
                     /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."
     promoter        complement(9930..11141)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(10074..10093)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     promoter        11154..11172
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     ncRNA           complement(11267..11393)
                     /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"
     ncRNA           complement(11421..11435)
                     /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"
     CDS             complement(12886..12909)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(12946..12969)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(12985..13008)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(13024..13047)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(13057..13081)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(13078..13116)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    13128..13146
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13164..13182
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13199..13217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13235..13253
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13271..13289
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13306..13324
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        13360..13407
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        13399..13417
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      13421..13430
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             13427..13801
                     /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"
     misc_feature    13866..13983
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa acctaccaag
       61 cctcctacta tcattatgaa taattttata taccacagcc aatttgttat gttaaaccaa
      121 ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt ttcttgctgg
      181 ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat ttctctgtcc
      241 cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact ccaactagca
      301 ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat ccccaggagc
      361 tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg cttgatgccc
      421 cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa attgttctgc
      481 tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat tgacgctgcg
      541 cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat tcccactgct
      601 cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc taatggttca
      661 atttttacta ctttatattt atataattca cttctccaat tgtccctcat atctcctcct
      721 ccaggtctga agatcagcgg ccggccgctt gctgtgcggt ggtcttactt ttgttttgct
      781 cttcctctat cttgtctaaa gcttccttgg tgtcttttat ctctatcctt tgatgcacac
      841 aatagagggt tgctactgta ttatataatg atctaagttc ttctgatcct gtctgaaggg
      901 atggttgtag ctgtcccagt atttgtctac agccttctga tgtttctaac aggccaggat
      961 taactgcgaa tcgttctagc tccctgcttg cccatactat atgttttaat ttatattttt
     1021 tctttccccc tggccttaac cgaatttttt cccatcgcga tctaattctc ccccgcttaa
     1081 tactgacgct ctcgcaccca tctctctcct tctagcctcc gctagtcaaa atttttggcg
     1141 tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc
     1201 gtcgagagag ctcctctggt ttccctttcg ctttcaagtc cctgttcggg cgccactgct
     1261 agagattttc cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa
     1321 cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg
     1381 gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt
     1441 acaggcaaaa agcagctgct tatatgcagg atctgagggc tcgccactcc ccagtcccgc
     1501 ccaggccacg cctccctgga aagtccccag cggaaagtcc cttgtagcaa gctcgatatc
     1561 agcagttctt gaagtactcc ggatgcagct ctcgggccac gtgatgaaat gctaggcggc
     1621 tgtcaaacct ccactctaac acttctctct ccgggtcatc catcccatgc aggctcacag
     1681 ggtgtaacaa gctggtgttc tctcctttat tggcctcttc taccttatct ggctcaactg
     1741 gtactagctt gtagcaccat ccaaaggtca gtggatatct gacccctggc cctggtgtgt
     1801 agttctgcta atcagggaag tagccttgtg tgtggtagat ccacagatca aggatatctt
     1861 gtcttctttg ggagtgaatt agcccttcca actactaagt ttgtagtaca tatttaacaa
     1921 atacaatttc tttaaaatga aaataattca gaggaatcac aggtttagag taaatgaaac
     1981 cacaggtaat tggcagtggt aatagggtat ggggtgggaa gtttgggatg attttggtta
     2041 gcttgagtta tccagttgat ccagacatga taagatacat tgatgagttt ggacaaacca
     2101 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2161 ttgtaaccat tataagctgc aataaacaag ttgatctccc gatccgtcga cgtcaggtgg
     2221 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2281 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2341 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2401 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2461 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2521 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2581 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2641 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2701 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2761 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2821 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2881 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2941 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3001 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3061 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3121 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3181 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3241 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3301 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3361 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3421 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3481 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3541 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3601 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3661 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3721 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3781 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3841 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3901 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3961 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4021 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4081 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4141 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     4201 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     4261 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     4321 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     4381 gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga gtttggacaa
     4441 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     4501 ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat
     4561 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
     4621 cctttcatag aaggcggcgg tggaatcgaa atctcgtagc acgtgctatt cctttgccct
     4681 cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt
     4741 ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg
     4801 gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa
     4861 gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc
     4921 tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca
     4981 cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct
     5041 ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc
     5101 cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag
     5161 agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg
     5221 gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg
     5281 tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc
     5341 ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac
     5401 accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattaat tccccaatgt
     5461 caagcacttc cggaatcggg agcgcggccg atgcaaagtg ccgataaaca taacgatctt
     5521 tgtagaaacc atcggcgcag ctatttaccc gcaggacata tccacgccct cctacatcga
     5581 agctgaaagc acgagattct tcgccctccg agagctgcat caggtcggag acgctgtcga
     5641 acttttcgat cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt ttcatcacgt
     5701 gctgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa aagcctaggc
     5761 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg gcctcggcct
     5821 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     5881 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     5941 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     6001 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     6061 ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa tcagctcatt
     6121 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     6181 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     6241 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     6301 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     6361 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     6421 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6481 acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga taccccctag agccccagct
     6541 ggttctttcc gcctcagaag ccatagagcc caccgcatcc ccagcatgcc tgctattgtc
     6601 ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct
     6661 actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc
     6721 ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg
     6781 cacagtcgag gctgatcagc gggtttaaac aatgtcaagg cctctcactc tctgatattc
     6841 atttctttgc aagttataaa tactgaataa taagatgaca tgaactacta ctgctagaga
     6901 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     6961 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     7021 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacaag
     7081 caaaaagcag atcttgtctt cgttgggagt gaattagccc ttccagtccc ctcttttctt
     7141 ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa ggtacctgag
     7201 gtgtgactgg aaaacccacc tcctcctcct cttgtgcttc tagccaggca caatcagcat
     7261 tggtagctgc tgtattgcta cttgtgattg ctccatgttt ttctaggtct cgactgcaga
     7321 attaattcca ggcggggagg cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg
     7381 aagacgcgga agaggccgca gagccggcag caggccgcgg gaaggaaggt ccgctggatt
     7441 gagggccgaa gggacgtagc agaaggacgt cccgcgcaga atccaggtgg caacacaggc
     7501 gagcagccat ggaaaggacg tcagcttccc cgacaacacc acggaattgt cagtgcccaa
     7561 cagccgagcc cctgtccagc agcgggcaag gcaggcggcg atgagttccg ccgtggcaat
     7621 agggaggggg aaagcgaaag tcccggaaag gagctgacag gtggtggcaa tgccccaacc
     7681 agtgggggtt gcgtcagcaa acacagtgca caccacgcca cgttgcctga caacgggcca
     7741 caactcctca taaagagaca gcaaccagga tttatacaag gaggagaaaa tgaaagccat
     7801 acgggaagca atagcatgat acaaaggcat taaagcagcg tatccacata gcgtaaaagg
     7861 agcaacatag ttaagaatac cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt
     7921 gttccagacg cgaactcagg caccgggctt gcgggtcatg caccaggtgc gcggtccttc
     7981 gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg
     8041 gggcgcggag gtctccagga aggcgggcac cccagcgcgc tcggccgcct ccactccggg
     8101 gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc
     8161 caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg
     8221 cgcggccagc cgggaaccgc tcaactcggc catgcgcgag ccgatctcgg cgaacaccgc
     8281 ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac
     8341 ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac
     8401 ccgctcgatg tggcggtccg ggtcgacggt gtggcgcgtg gcggggtagt cggcgaacgc
     8461 ggcggcgagg gtgcgtacgg cccgggggac gtcgtcgcgg gtggcgaggc gcaccgtggg
     8521 cttgtactcg gtggccatgg tattatcgtg tttttcaaag gaaaaccacg tccccgtggt
     8581 tcggggggcc tagacgtttt tttaacctcg actaaacaca tgtaaagcat gtgcaccgag
     8641 gccccagatc agatcccata caatggggta ccttctgggc atccttcagc cccttgttga
     8701 atacgcttga ggagagccat ttgactcttt ccacaactat ccaactcaca acgtggcact
     8761 ggggttgtgc cgcctttgca ggtgtatctt atacacgtgg cttttggccg cagaggcacc
     8821 tgtcgccagg tggggggttc cgctgcctgc aaagggtcgc tacagacgtt gtttgtcttc
     8881 aagaagcttc cagaggaact gcttccttca cgacattcaa cagaccttgc attcctttgg
     8941 cgagagggga aagaccccta ggaatgctcg tcaagaagac agggccaggt ttccgggccc
     9001 tcacattgcc aaaagacggc aatatggtgg aaaataacat atagacaaac gcacaccggc
     9061 cttattccaa gcggcttcgg ccagtaacgt tagggggggg ggagggagag gggcggaatt
     9121 tgcggccgcg gatccataac ttcgtataat gtatactata cgaagttatt tacccgggga
     9181 gcatgtcaag gtcaaaatcg tcaagggcat cggctgggag catgtctaag tcaaaatcgt
     9241 caagggcgtc ggtcggcccg ccgctttcgc actttagctg tttctccagg ccacatatga
     9301 ttagttccag gccgaaaagg aaggcaggtt cggctccctg ccggtcgaac agctcaattg
     9361 cttgtctcag aagtgggggc atagaatcgg tggtaggtgt ctctctttcc tcttttgcta
     9421 cttgatgctc ctgttcctcc aatacgcagc ccagtgtaaa gtggcccacg gcggacagag
     9481 cgtacagtgc gttctccagg gagaagcctt gctgacacag gaacgcgagc tgattttcca
     9541 gggtttcgta ctgtttctct gttgggcggg tgccgagatg cactttagcc ccgtcgcgat
     9601 gtgagaggag agcacagcgg tatgacttgg cgttgttccg cagaaagtct tgccatgact
     9661 cgccttccag ggggcagaag tgggtatgat gcctgtccag catctcgatt ggcagggcat
     9721 cgagcagggc ccgcttgttc ttcacgtgcc agtacagggt aggctgctca actcccagct
     9781 tttgagcgag tttccttgtc gtcaggcctt cgataccgac accattgagt aattccagag
     9841 ctccgtttat gactttgctc ttgtccagcc tagacatggt ggcataactt cgtataatgt
     9901 atactatacg aagttatgga tccgctagcg tctaacaaaa aagccaaaaa cggccagaat
     9961 ttagcggaca atttactagt ctaacactga aaattacata ttgacccaaa tgattacatt
    10021 tcaaaaggtg cctaaaaaac ttcacaaaac acactcgcca accccgagcg catagttcaa
    10081 aaccggagct tcagctactt aagaagatag gtacataaaa ccgaccaaag aaactgacgc
    10141 ctcacttatc cctcccctca ccagaggtcc ggcgcctgtc gattcaggag agcctaccct
    10201 aggcccgaac cctgcgtcct gcgacggaga aaagcctacc gcacacctac cggcaggtgg
    10261 ccccaccctg cattataagc caacagaacg ggtgacgtca cgacacgacg agggcgcgcg
    10321 ctcccaaagg tacgggtgca ctgcccaacg gcaccgccat aactgccgcc cccgcaacag
    10381 acgacaaacc gagttctcca gtcagtgaca aacttcacgt cagggtcccc agatggtgcc
    10441 ccagcccatc tcacccgaat aagagctttc ccgcattagc gaaggcctca agaccttggg
    10501 ttcttgccgc ccaccatgcc ccccaccttg tttcaacgac ctcacagccc gcctcacaag
    10561 cgtcttccat tcaagactcg ggaacagccg ccattttgct gcgctccccc caacccccag
    10621 ttcagggcaa ccttgctcgc ggacccagac tacagccctt ggcggtctct ccacacgctt
    10681 ccgtcccacc gagcggcccg gcggccacga aagccccggc cagcccagca gcccgctact
    10741 caccaagtga cgatcacagc gatccacaaa caagaaccgc gacccaaatc ccggctgcga
    10801 cggaactagc tgtgccacac ccggcgcgtc cttatataat catcggcgtt caccgcccca
    10861 cggagatccc tccgcagaat cgccgagaag ggactacttt tcctcgcctg ttccgctctc
    10921 tggaaagaaa accagtgccc tagagtcacc caagtcccgt cctaaaatgt ccttctgctg
    10981 atactggggt tctaaggccg agtcttatga gcagcgggcc gctgtcctga gcgtccgggc
    11041 ggaaggatca ggacgctcgc tgcgcccttc gtctgacgtg gcagcgctcg ccgtgaggag
    11101 gggggcgccc gcgggaggcg ccaaaacccg gcgcggaggc cacgcgtcct aggtaatacg
    11161 actcactata gggcccgccc gcaaacaaca tcaaacaaac cagcagacat atgcaacaag
    11221 acacgagacg gagtaatggc cggccgcatt agtcttccaa ttgaaaaaag tgatttaatt
    11281 tataccattt taattcagct ttgtaaaaat gtatcaaaga gatagcaagg tattcagttt
    11341 tagtaaacaa gataattgct cctaaagtag ccccttgaat tccgaggcag taggcactct
    11401 cgcttgggcg agagtaagta catctgtggc ttcactactt actctcgccc aagcgagatc
    11461 gctcactgtc aacagcaata taccttctca actacacatg aagtcctcca ccagaggcag
    11521 cttttccaaa tcataggcat cgaagaggtc actgatgccc tcttcctccc ccaggctcag
    11581 caggtagtcc tcttggagca ggggaggcag taagttcaca aaaggtcctt caaggttgga
    11641 cgggatttgg tcctcagtct gctgtaaaag gttggctggg gaggccagag gagagaggtt
    11701 tgctgtagaa accgagcagt cactatgtcc tgagttgtta gaagccaagt ctttggaagt
    11761 gggcttgggg atattcccat tgtggtcttg gttatttgtt ttcatgggtc tgtgtgtttc
    11821 cgtctcttct ggacacaagt aaacctcaat gggcccttgg gtacttgcca aatggatttg
    11881 taggctctct attgagtcag gcacttcaag tctcgtttct ggaggggctt tcacaactat
    11941 aacagtttgg tctttaaggc cactaatttt tcgaatatct tgatatgtaa cataagctaa
    12001 cctttgattc tctgaatcct cggttaacag tttgaggtcc agggtacagc tttggatcag
    12061 ctcatctaat ttcttctctt cctgactgag ctcagtcact tctttggaca ggccttgaca
    12121 ctgggccagc atgcccccat cctcagacag actgcagccc atccactgga cgttgttctt
    12181 agacttcttc ttaatgaggt ggatgccttc cagcacgttg gtgatgtcgt aaatcctcct
    12241 cttctgcacc ttgagcacct ctgctgcctt gttcagatcc aggaccccat caggagactg
    12301 gctcaggagc tgaatgaact tcttggtgag cagaccgagg gacgtatcat accgcgtttt
    12361 ttctgaggga gattttggag tttttggact atcgggactc cgtagtgcag ctcttccttt
    12421 gcccttgggg gtctttagac catctgagag gtactgatgg ccactctcgc ccagttccag
    12481 ccttcgcttt gccggtgggc caccgccgcc gccgcgtcct ggtgctggtg gctgctgcag
    12541 cagcccgact ctgctggagg gtccttgtgg cgtggtgtag aggctgctgg cggccggctc
    12601 cgtgccgggg acactgggga gaagggggcc ggcggtcagg gcgccacttt ggagggaggt
    12661 ggcacaggac gtggtggaag ggttcgtagt gaggatctgg atgtacgtgc ccggggcggc
    12721 ggcggcggcg aagccggggc tggctagcag tgcccttttg tccatggagg ctgcagcggc
    12781 ggcggcgacg acagccgccc cctccccacc cccggcggtc accaggtact gctccagggc
    12841 gggctggatc ccctttctga attccaccac actggatcca gctcctttct cgaactgcgg
    12901 gtggctccag cttcctcctc cgctgcctcc gccgcttcct cctcccttct cgaactgagg
    12961 gtggctccag ctggcggccg accctttatc atcatcatct ttataatcca tagatcctgg
    13021 taccttgtca tcgtcgtcct tgtagtccat ggttgcgatc tgacggttca ctaaacgagc
    13081 tctgcttata taggcctccc accgtacacg cctacctcga catacgttct ctatcactga
    13141 tagggagtaa actcgacata cgttctctat cactgatagg gataaactcg acatacgttc
    13201 tctatcactg atagggagta aactcgacat acgttctcta tcactgatag ggagtaaact
    13261 cgacatacgt tctctatcac tgatagggag taaactcgac atcgttctct atcactgata
    13321 gggagtaaac tcgtctagaa cctcggacct gcagcacgtg ttgacaatta atcatcggca
    13381 tagtatatcg gcatagtata atacgactca ctataggagg gccaccatgg ccaagttgac
    13441 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
    13501 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
    13561 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
    13621 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
    13681 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
    13741 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
    13801 attccggact agagcgggcc cggtactaac caaactggat ctctgctgtc cctgtaataa
    13861 acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt
    13921 gctattatgt ctactattct ttcccctgca ctgtaccccc caatcccccc ttttctttta
    13981 aaattgtgga tgaatactgc catttgtctg cagaattggc gcacgcagtg ccgatccgtt
    14041 cactaatcga atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg
    14101 cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatat
//
