LOCUS       sequence-473634-       14080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  doxycycline inducible expression of E2F1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14080)
  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 14080)
  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-E2F1
FEATURES             Location/Qualifiers
     source          1..14080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(88..132)
                     /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    317..550
                     /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    1047..1172
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1219..1852
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    2020..2154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2202..2306
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2307..2375,2376..3167)
                     /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(2525..2544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3338..3926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3827..3846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4080..4097
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4214..4235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4250..4255,4256..4273,4274..4280)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4288..4304
                     /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     4293..4315
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4312..4328
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4312..4328
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4377..4498
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(5706..6035)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5720..5855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5774..5793)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6020..6040
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6049..6477)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6159..6180)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6371..6390
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6523..6747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6736..6753
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6854..7087
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    7294..7882
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7400..7411
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7809..7829
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7898..8500)
                     /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(7981..8001)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8503..9075
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8662..8681)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     8891..8908
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    9098..9131
                     /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(9132..9839)
                     /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    9846..9879
                     /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(9892..11103)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(10036..10055)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     promoter        11116..11134
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     ncRNA           complement(11229..11355)
                     /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(11383..11397)
                     /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(12773..12796)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(12833..12856)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(12872..12895)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(12911..12934)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(12944..12968)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(12965..13003)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    13015..13033
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13051..13069
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13086..13104
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13122..13140
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13158..13176
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13193..13211
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        13247..13294
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        13286..13304
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      13308..13317
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             13314..13688
                     /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    13753..13870
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 aactattctt aaacctacca agcctcctac tatcattatg aataatttta tataccacag
       61 ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc
      121 ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt
      181 gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag
      241 agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga
      301 gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct
      361 tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat
      421 agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac
      481 cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa
      541 caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt
      601 gggtgctact cctaatggtt caatttttac tactttatat ttatataatt cacttctcca
      661 attgtccctc atatctcctc ctccaggtct gaagatcagc ggccggccgc ttgctgtgcg
      721 gtggtcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
      781 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
      841 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
      901 gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct tgcccatact
      961 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
     1021 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
     1081 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
     1141 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
     1201 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     1261 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     1321 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     1381 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
     1441 gctcgccact ccccagtccc gcccaggcca cgcctccctg gaaagtcccc agcggaaagt
     1501 cccttgtagc aagctcgata tcagcagttc ttgaagtact ccggatgcag ctctcgggcc
     1561 acgtgatgaa atgctaggcg gctgtcaaac ctccactcta acacttctct ctccgggtca
     1621 tccatcccat gcaggctcac agggtgtaac aagctggtgt tctctccttt attggcctct
     1681 tctaccttat ctggctcaac tggtactagc ttgtagcacc atccaaaggt cagtggatat
     1741 ctgacccctg gccctggtgt gtagttctgc taatcaggga agtagccttg tgtgtggtag
     1801 atccacagat caaggatatc ttgtcttctt tgggagtgaa ttagcccttc caactactaa
     1861 gtttgtagta catatttaac aaatacaatt tctttaaaat gaaaataatt cagaggaatc
     1921 acaggtttag agtaaatgaa accacaggta attggcagtg gtaatagggt atggggtggg
     1981 aagtttggga tgattttggt tagcttgagt tatccagttg atccagacat gataagatac
     2041 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2101 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttgatctc
     2161 ccgatccgtc gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2221 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2281 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2341 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2401 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2461 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2521 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2581 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2641 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2701 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2761 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2821 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2881 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2941 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3001 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3061 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3121 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3181 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3241 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3301 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3421 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3481 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3541 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3601 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3661 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3721 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3781 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3841 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3901 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3961 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4021 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4081 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     4141 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     4201 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     4261 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     4321 gctatgacca tgattacgcc aagctctagc tagaggtcga cggtatacag acatgataag
     4381 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     4441 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg
     4501 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     4561 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgta
     4621 gcacgtgcta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca ctatcggcga
     4681 gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc
     4741 cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc caagctgcat
     4801 catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg cggagcatat
     4861 acgcccggag ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct
     4921 gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc tcgtattggg
     4981 aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca
     5041 ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg
     5101 cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg tcgtccatca
     5161 cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca cgccatgtag
     5221 tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg ctaagatcgg
     5281 ccgcagcgat cgcatccatg gcctccgcga ccggctgcag aacagcgggc agttcggttt
     5341 caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag gtcaggctct
     5401 cgctgaatta attccccaat gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag
     5461 tgccgataaa cataacgatc tttgtagaaa ccatcggcgc agctatttac ccgcaggaca
     5521 tatccacgcc ctcctacatc gaagctgaaa gcacgagatt cttcgccctc cgagagctgc
     5581 atcaggtcgg agacgctgtc gaacttttcg atcagaaact tctcgacaga cgtcgcggtg
     5641 agttcaggct ttttcatcac gtgctgatca gatccgaaaa tggatataca agctcccggg
     5701 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     5761 gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     5821 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     5881 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     5941 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     6001 gagcctgggg actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa
     6061 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     6121 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     6181 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     6241 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     6301 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     6361 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     6421 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtggg
     6481 gataccccct agagccccag ctggttcttt ccgcctcaga agccatagag cccaccgcat
     6541 ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
     6601 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     6661 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     6721 cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttaa acaatgtcaa
     6781 ggcctctcac tctctgatat tcatttcttt gcaagttata aatactgaat aataagatga
     6841 catgaactac tactgctaga gattttccac actgactaaa agggtctgag ggatctctag
     6901 ttaccagagt cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat
     6961 tgaggcttaa gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct
     7021 aaccagagag acccagtaca agcaaaaagc agatcttgtc ttcgttggga gtgaattagc
     7081 ccttccagtc ccctcttttc ttttaaaaag tggctaagat ctacagctgc cttgtaagtc
     7141 attggtctta aaggtacctg aggtgtgact ggaaaaccca cctcctcctc ctcttgtgct
     7201 tctagccagg cacaatcagc attggtagct gctgtattgc tacttgtgat tgctccatgt
     7261 ttttctaggt ctcgactgca gaattaattc caggcgggga ggcggcccaa agggagatcc
     7321 gactcgtctg agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc
     7381 gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca
     7441 gaatccaggt ggcaacacag gcgagcagcc atggaaagga cgtcagcttc cccgacaaca
     7501 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     7561 cgatgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac
     7621 aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc
     7681 cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca
     7741 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     7801 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     7861 ttttgtaatc cagaggttga ttgttccaga cgcgaactca ggcaccgggc ttgcgggtca
     7921 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
     7981 gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccagcgc
     8041 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
     8101 cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     8161 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     8221 agccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     8281 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     8341 agagttcttg cagctcggtg acccgctcga tgtggcggtc cgggtcgacg gtgtggcgcg
     8401 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggcccggggg acgtcgtcgc
     8461 gggtggcgag gcgcaccgtg ggcttgtact cggtggccat ggtattatcg tgtttttcaa
     8521 aggaaaacca cgtccccgtg gttcgggggg cctagacgtt tttttaacct cgactaaaca
     8581 catgtaaagc atgtgcaccg aggccccaga tcagatccca tacaatgggg taccttctgg
     8641 gcatccttca gccccttgtt gaatacgctt gaggagagcc atttgactct ttccacaact
     8701 atccaactca caacgtggca ctggggttgt gccgcctttg caggtgtatc ttatacacgt
     8761 ggcttttggc cgcagaggca cctgtcgcca ggtggggggt tccgctgcct gcaaagggtc
     8821 gctacagacg ttgtttgtct tcaagaagct tccagaggaa ctgcttcctt cacgacattc
     8881 aacagacctt gcattccttt ggcgagaggg gaaagacccc taggaatgct cgtcaagaag
     8941 acagggccag gtttccgggc cctcacattg ccaaaagacg gcaatatggt ggaaaataac
     9001 atatagacaa acgcacaccg gccttattcc aagcggcttc ggccagtaac gttagggggg
     9061 ggggagggag aggggcggaa tttgcggccg cggatccata acttcgtata atgtatacta
     9121 tacgaagtta tttacccggg gagcatgtca aggtcaaaat cgtcaagggc atcggctggg
     9181 agcatgtcta agtcaaaatc gtcaagggcg tcggtcggcc cgccgctttc gcactttagc
     9241 tgtttctcca ggccacatat gattagttcc aggccgaaaa ggaaggcagg ttcggctccc
     9301 tgccggtcga acagctcaat tgcttgtctc agaagtgggg gcatagaatc ggtggtaggt
     9361 gtctctcttt cctcttttgc tacttgatgc tcctgttcct ccaatacgca gcccagtgta
     9421 aagtggccca cggcggacag agcgtacagt gcgttctcca gggagaagcc ttgctgacac
     9481 aggaacgcga gctgattttc cagggtttcg tactgtttct ctgttgggcg ggtgccgaga
     9541 tgcactttag ccccgtcgcg atgtgagagg agagcacagc ggtatgactt ggcgttgttc
     9601 cgcagaaagt cttgccatga ctcgccttcc agggggcaga agtgggtatg atgcctgtcc
     9661 agcatctcga ttggcagggc atcgagcagg gcccgcttgt tcttcacgtg ccagtacagg
     9721 gtaggctgct caactcccag cttttgagcg agtttccttg tcgtcaggcc ttcgataccg
     9781 acaccattga gtaattccag agctccgttt atgactttgc tcttgtccag cctagacatg
     9841 gtggcataac ttcgtataat gtatactata cgaagttatg gatccgctag cgtctaacaa
     9901 aaaagccaaa aacggccaga atttagcgga caatttacta gtctaacact gaaaattaca
     9961 tattgaccca aatgattaca tttcaaaagg tgcctaaaaa acttcacaaa acacactcgc
    10021 caaccccgag cgcatagttc aaaaccggag cttcagctac ttaagaagat aggtacataa
    10081 aaccgaccaa agaaactgac gcctcactta tccctcccct caccagaggt ccggcgcctg
    10141 tcgattcagg agagcctacc ctaggcccga accctgcgtc ctgcgacgga gaaaagccta
    10201 ccgcacacct accggcaggt ggccccaccc tgcattataa gccaacagaa cgggtgacgt
    10261 cacgacacga cgagggcgcg cgctcccaaa ggtacgggtg cactgcccaa cggcaccgcc
    10321 ataactgccg cccccgcaac agacgacaaa ccgagttctc cagtcagtga caaacttcac
    10381 gtcagggtcc ccagatggtg ccccagccca tctcacccga ataagagctt tcccgcatta
    10441 gcgaaggcct caagaccttg ggttcttgcc gcccaccatg ccccccacct tgtttcaacg
    10501 acctcacagc ccgcctcaca agcgtcttcc attcaagact cgggaacagc cgccattttg
    10561 ctgcgctccc cccaaccccc agttcagggc aaccttgctc gcggacccag actacagccc
    10621 ttggcggtct ctccacacgc ttccgtccca ccgagcggcc cggcggccac gaaagccccg
    10681 gccagcccag cagcccgcta ctcaccaagt gacgatcaca gcgatccaca aacaagaacc
    10741 gcgacccaaa tcccggctgc gacggaacta gctgtgccac acccggcgcg tccttatata
    10801 atcatcggcg ttcaccgccc cacggagatc cctccgcaga atcgccgaga agggactact
    10861 tttcctcgcc tgttccgctc tctggaaaga aaaccagtgc cctagagtca cccaagtccc
    10921 gtcctaaaat gtccttctgc tgatactggg gttctaaggc cgagtcttat gagcagcggg
    10981 ccgctgtcct gagcgtccgg gcggaaggat caggacgctc gctgcgccct tcgtctgacg
    11041 tggcagcgct cgccgtgagg aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag
    11101 gccacgcgtc ctaggtaata cgactcacta tagggcccgc ccgcaaacaa catcaaacaa
    11161 accagcagac atatgcaaca agacacgaga cggagtaatg gccggccgca ttagtcttcc
    11221 aattgaaaaa agtgatttaa tttataccat tttaattcag ctttgtaaaa atgtatcaaa
    11281 gagatagcaa ggtattcagt tttagtaaac aagataattg ctcctaaagt agccccttga
    11341 attccgaggc agtaggcact ctcgcttggg cgagagtaag tacatctgtg gcttcactac
    11401 ttactctcgc ccaagcgaga tcgctcactg tcaacagcaa tataccttct cagaaatcca
    11461 ggggggtgag gtccccaaag tcacagtcga agaggtctct gatgccctcg ccctcctcga
    11521 ggccgaagtg gtagtcgagg gcctcgtggg gtggggaaag gctgatgaac tcctcaggga
    11581 ggaggccgga gaagtcctcc cgcacatgct ccaggagcga gtcggccgcc accagcgggg
    11641 acaggcggtc ctcgtccacg ggagcccgca ggctgcccat ccgggacaac agcggttttt
    11701 gctccaggct gagtagagac tggctgggat ctgtggtgag ggatgagggg ggagatgatg
    11761 gtggtggtga cactatggtg gcagagtcag tggccctgtt ctcctcctca gaagtgacct
    11821 cctgggatgg ggtcttccca gggctgatcc cacctacggt ctcctcaggg cacaggaaaa
    11881 catcgatcgg gccttgtttg ctcttaaggg agatctgaaa gttctccgaa gagtccacgg
    11941 cttggagctg ggtctcagga ggggctttga tcaccataac catctgctct gcagggtctg
    12001 caatgctacg aaggtcctga cacgtcacgt aggccaggcg ctggctgtca gtgtcctcgg
    12061 agagcaggcg cagctgcgta gtacagatat tcatcaggtg gtccagctgc tgctcgctct
    12121 cctgcagctg tcggaggtcc tgggtcaacc cctcaagccg tccgccgacg cccactgtgg
    12181 tgtggctgcc cagccactgg atgtggttct tggacttctt ggcaatgagc tggatgccct
    12241 caaggacgtt ggtgatgtca tagatgcgcc gcttctgcac cttcagcacc tcggcagccc
    12301 agttcaggtc gacgacaccg tcagccgagt ggctcagcag ctccaggaag cgcttggtgg
    12361 tcagattcag tgaggtctca tagcgtgact tctcccccgg ggatttcaca ccttttcctg
    12421 gatggcggcc tctgccccga gctggcccac tgctctcggc caggtactga tggtcagttt
    12481 ccaggtccag cctccgcttc accggcgggc ggccgagcgc gggccgcggc gcactgggtg
    12541 tgggccgggg cgcctgcggt gtggcgaaga gcagcaggtc agggtcgcag gggccggcgg
    12601 cgggcgccgc ggggccggtg ggagccggcg gggcgctggc gtcctgcgcg gcggagatga
    12661 tgacgatctg cgaggagtcg agcagccgca gcgcgccggc cccgagcagg gcctccagcg
    12721 ccggcgcgca tgggccgccc gcaggggccc cggccaaggc ggatccagct cctttctcga
    12781 actgcgggtg gctccagctt cctcctccgc tgcctccgcc gcttcctcct cccttctcga
    12841 actgagggtg gctccagctg gcggccgacc ctttatcatc atcatcttta taatccatag
    12901 atcctggtac cttgtcatcg tcgtccttgt agtccatggt tgcgatctga cggttcacta
    12961 aacgagctct gcttatatag gcctcccacc gtacacgcct acctcgacat acgttctcta
    13021 tcactgatag ggagtaaact cgacatacgt tctctatcac tgatagggat aaactcgaca
    13081 tacgttctct atcactgata gggagtaaac tcgacatacg ttctctatca ctgataggga
    13141 gtaaactcga catacgttct ctatcactga tagggagtaa actcgacatc gttctctatc
    13201 actgataggg agtaaactcg tctagaacct cggacctgca gcacgtgttg acaattaatc
    13261 atcggcatag tatatcggca tagtataata cgactcacta taggagggcc accatggcca
    13321 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
    13381 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
    13441 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
    13501 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
    13561 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
    13621 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
    13681 aggactgatt ccggactaga gcgggcccgg tactaaccaa actggatctc tgctgtccct
    13741 gtaataaacc cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat
    13801 gtctgttgct attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt
    13861 tcttttaaaa ttgtggatga atactgccat ttgtctgcag aattggcgca cgcagtgccg
    13921 atccgttcac taatcgaatg gatctgtctc tgtctctctc tccaccttct tcttctattc
    13981 cttcgggcct gtcgggtccc ctcggggttg ggaggtgggt ctgaaacgat aatggtgaat
    14041 atccctgcct aactctattc actatagaaa gtacagcaaa
//
