LOCUS       sequence-420490-       10471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express tagged hRb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10471)
  AUTHORS   Konagaya Y, Rosenthal D, Ratnayeke N, Fan Y, Meyer T
  TITLE     An intermediate Rb-E2F activity state safeguards proliferation 
            commitment.
  JOURNAL   Nature. 2024 Jul;631(8020):424-431. doi: 10.1038/s41586-024-07554-2.
            Epub 2024 Jun 26.
  PUBMED    38926571
REFERENCE   2  (bases 1 to 10471)
  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: 
            "10.1038/s41586-024-07554-2"; journalName: "Nature"; date: 
            "2024-07"; volume: "631"; issue: "8020"; pages: "424-431"
COMMENT     Sequence Label: pCW-HA-hRb-WT-puro
FEATURES             Location/Qualifiers
     source          1..10471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        212..526
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    220..238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    256..274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    291..309
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    327..345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    363..381
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    398..416
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    434..452
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     499..523
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             552..578
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        3413..3923
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3842..3860
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3933..4529
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3933..3952)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4429..4449
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4530..4583
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4584..5201,5202..5207,5208..5243,5244..5246,5247..5282,
                     5283..5285,5286..5321,5322..5330)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(5370..5390)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5788..5799)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6004..6237
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6309..6430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6346..6365)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6400..6419
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6470..6605
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6605..6676
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6677..6745,6746..7537)
                     /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(6895..6914)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7708..8296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8197..8216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8450..8467
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8584..8605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8620..8625,8626..8643,8644..8650)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8658..8674
                     /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     8663..8685
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8682..8698
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8682..8698
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8717..8737
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8719..8737
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8765..8991
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8992..9172
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9219..9344
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9837..10070
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10255..10299
                     /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"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      541 gcgccaccat gtacccatac gatgttccag attacgctct tatggccatg gaggcccgaa
      601 ttcggtcgac cccgcccaaa accccccgaa aaacggccgc caccgccgcc gctgccgccg
      661 cggaaccccc ggcaccgccg ccgccgcccc ctcctgagga ggacccagag caggacagcg
      721 gcccggagga cctgcctctc gtcaggcttg agtttgaaga aacagaagaa cctgatttta
      781 ctgcattatg tcagaaatta aagataccag atcatgtcag agagagagct tggttaactt
      841 gggagaaagt ttcatctgtg gatggagtat tgggaggtta tattcaaaag aaaaaggaac
      901 tgtggggaat ctgtatcttt attgcagcag ttgacctaga tgagatgtcg ttcactttta
      961 ctgagctaca gaaaaacata gaaatcagtg tccataaatt ctttaactta ctaaaagaaa
     1021 ttgataccag taccaaagtt gataatgcta tgtcaagact gttgaagaag tatgatgtat
     1081 tgtttgcact cttcagcaaa ttggaaagga catgtgaact tatatatttg acacaaccca
     1141 gcagttcgat atctactgaa ataaattctg cattggtgct aaaagtttct tggatcacat
     1201 ttttattagc taaaggggaa gtattacaaa tggaagatga tctggtgatt tcatttcagt
     1261 taatgctatg tgtccttgac tattttatta aactctcacc tcccatgttg ctcaaagaac
     1321 catataaaac agctgttata cccattaatg gttcacctcg aacacccagg cgaggtcaga
     1381 acaggagtgc acggatagca aaacaactag aaaatgatac aagaattatt gaagttctct
     1441 gtaaagaaca tgaatgtaat atagatgagg tgaaaaatgt ttatttcaaa aattttatac
     1501 cttttatgaa ttctcttgga cttgtaacat ctaatggact tccagaggtt gaaaatcttt
     1561 ctaaacgata cgaagaaatt tatcttaaaa ataaagatct agatgcaaga ttatttttgg
     1621 atcatgataa aactcttcag actgattcta tagacagttt tgaaacacag agaacaccac
     1681 gaaaaagtaa ccttgatgaa gaggtgaatg taattcctcc acacactcca gttaggactg
     1741 ttatgaacac tatccaacaa ttaatgatga ttttaaattc agcaagtgat caaccttcag
     1801 aaaatctgat ttcctatttt aacaactgca cagtgaatcc aaaagaaagt atactgaaaa
     1861 gagtgaagga tataggatac atctttaaag agaaatttgc taaagctgtg ggacagggtt
     1921 gtgtcgaaat tggatcacag cgatacaaac ttggagttcg cttgtattac cgagtaatgg
     1981 aatccatgct taaatcagaa gaagaacgat tatccattca aaattttagc aaacttctga
     2041 atgacaacat ttttcatatg tctttattgg cgtgcgctct tgaggttgta atggccacat
     2101 atagcagaag tacatctcag aatcttgatt ctggaacaga tttgtctttc ccatggattc
     2161 tgaatgtgct taatttaaaa gcctttgatt tttacaaagt gatcgaaagt tttatcaaag
     2221 cagaaggcaa cttgacaaga gaaatgataa aacatttaga acgatgtgaa catcgaatca
     2281 tggaatccct tgcatggctc tcagattcac ctttatttga tcttattaaa caatcaaagg
     2341 accgagaagg accaactgat caccttgaat ctgcttgtcc tcttaatctt cctctccaga
     2401 ataatcacac tgcagcagat atgtatcttt ctcctgtaag atctccaaag aaaaaaggtt
     2461 caactacgcg tgtaaattct actgcaaatg cagagacaca agcaacctca gccttccaga
     2521 cccagaagcc attgaaatct acctctcttt cactgtttta taaaaaagtg tatcggctag
     2581 cctatctccg gctaaataca ctttgtgaac gccttctgtc tgagcaccca gaattagaac
     2641 atatcatctg gacccttttc cagcacaccc tgcagaatga gtatgaactc atgagagaca
     2701 ggcatttgga ccaaattatg atgtgttcca tgtatggcat atgcaaagtg aagaatatag
     2761 accttaaatt caaaatcatt gtaacagcat acaaggatct tcctcatgct gttcaggaga
     2821 cattcaaacg tgttttgatc aaagaagagg agtatgattc tattatagta ttctataact
     2881 cggtcttcat gcagagactg aaaacaaata ttttgcagta tgcttccacc aggcccccta
     2941 ccttgtcacc aatacctcac attcctcgaa gcccttacaa gtttcctagt tcacccttac
     3001 ggattcctgg agggaacatc tatatttcac ccctgaagag tccatataaa atttcagaag
     3061 gtctgccaac accaacaaaa atgactccaa gatcaagaat cttagtatca attggtgaat
     3121 cattcgggac ttctgagaag ttccagaaaa taaatcagat ggtatgtaac agcgaccgtg
     3181 tgctcaaaag aagtgctgaa ggaagcaacc ctcctaaacc actgaaaaaa ctacgctttg
     3241 atattgaagg atcagatgaa gcagatggaa gtaaacatct cccaggagag tccaaatttc
     3301 agcagaaact ggcagaaatg acttctactc gaacacgaat gcaaaagcag aaaatgaatg
     3361 atagcatgga tacctcaaac aaggaagaga aatgaaccgg tgaattcgat ccggggttgg
     3421 ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg
     3481 gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca
     3541 gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc
     3601 ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg
     3661 cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac
     3721 cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg
     3781 aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc
     3841 ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc
     3901 gaatcaccga cctctctccc cagcaattca ccatgaccga gtacaagccc acggtgcgcc
     3961 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     4021 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     4081 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     4141 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     4201 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     4261 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     4321 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     4381 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     4441 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     4501 cctggtgcat gacccgcaag cccggtgccg aaggtagagg ttctctcctc acttgtggtg
     4561 atgttgaaga aaaccctggt ccaatgtcta gactggacaa gagcaaagtc ataaacggag
     4621 ctctggaatt actcaatggt gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa
     4681 agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg gccctgctcg
     4741 atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc ctggaaggcg
     4801 agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct ctcctctcac
     4861 atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag tacgaaaccc
     4921 tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac gcactgtacg
     4981 ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag gagcatcaag
     5041 tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt ctgagacaag
     5101 caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc ctggaactaa
     5161 tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg
     5221 acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac cttgatatgc
     5281 tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa ctaagtaagg
     5341 atcgatccaa gatatcgtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5401 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5461 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5521 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5581 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5641 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5701 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     5761 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5821 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5881 atctcccttt gggccgcctc cccgcctgtt tcgcctcggc gtccggacta gaggtacctt
     5941 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     6001 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     6061 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6121 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6181 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6241 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     6301 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6361 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     6421 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     6481 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6541 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     6601 aggcctttca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6661 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6721 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6781 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6841 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6901 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     6961 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7021 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7081 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7141 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7201 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7261 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7321 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7381 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7441 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7501 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7561 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7621 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7681 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7741 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7801 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7861 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7921 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     7981 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8041 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8101 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     8161 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     8221 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8281 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8341 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     8401 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     8461 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     8521 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     8581 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     8641 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     8701 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
     8761 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
     8821 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
     8881 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
     8941 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
     9001 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     9061 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     9121 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     9181 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     9241 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     9301 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     9361 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     9421 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     9481 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     9541 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     9601 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     9661 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     9721 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     9781 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     9841 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     9901 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     9961 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
    10021 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
    10081 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
    10141 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
    10201 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
    10261 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
    10321 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
    10381 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
    10441 tttcagaccc acctcccaac cccgagggga c
//
