LOCUS       sequence-229486-       12299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a puromycin resistance cassette with Tet-on 3G from the 
            EF1Alpha promoter and Ascl1 from the TRE3G promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12299)
  AUTHORS   Liu Y, Yu C, Daley TP, Wang F, Cao WS, Bhate S, Lin X, Still C 2nd, 
            Liu H, Zhao D, Wang H, Xie XS, Ding S, Wong WH, Wernig M, Qi LS
  TITLE     CRISPR Activation Screens Systematically Identify Factors that Drive
            Neuronal Fate and Reprogramming.
  JOURNAL   Cell Stem Cell. 2018 Sep 28. pii: S1934-5909(18)30440-5. doi: 
            10.1016/j.stem.2018.09.003.
  PUBMED    30318302
REFERENCE   2  (bases 1 to 12299)
  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: "Cell Stem 
            Cell. 2018 Sep 28. pii: S1934-5909(18)30440-5. doi: 
            10.1016/j.stem.2018.09.003."
COMMENT     Sequence Label: pHR-EF1Alpha-puro-T2A-Tet-on 3G-TRE3G-Ascl1
FEATURES             Location/Qualifiers
     source          1..12299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             174..807
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    854..979
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1476..1709
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1894..1938
                     /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    2223..2340
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     intron          2568..3490
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3447..3467
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3578..4174
                     /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(3578..3597)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4074..4094
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4184..4237
                     /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             4250..4996
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    5034..5622
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5087..5107)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5505..5516)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    5667..5685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5703..5721
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5739..5757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5775..5793
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5811..5829
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5847..5865
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5883..5901
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5946..5970
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      6054..6063
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    6796..6917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6927..6943)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6928..6944)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7034..7267
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(7292..7310)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7293..7310)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7600..7619)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7833..7852)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7952..7974
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8012..8030)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8098..8202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8203..8271,8272..9063)
                     /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(8421..8440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9234..9822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9723..9742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9976..9993
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(10063..10083)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10068..10397
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10248..10383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          11531..11596
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             11726..11746
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    join(12171..12299,1..6)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
       61 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
      121 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg
      181 gctaattcac tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg
      241 ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt
      301 tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg
      361 agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
      421 agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca
      481 tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga
      541 ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat
      601 ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
      661 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
      721 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
      781 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
      841 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
      901 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
      961 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     1021 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1081 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1141 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1201 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1261 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1321 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     1381 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     1441 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     1501 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     1561 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     1621 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     1681 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     1741 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     1801 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     1861 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     1921 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     1981 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     2041 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     2101 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     2161 agatccattc gattagtgaa cggatctcga cggtatcgcc aaatggcagt attcatccac
     2221 aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     2281 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     2341 cgggtttatt acagggacag cagagatcca gtttggatcg ataagcttga tatcgaattc
     2401 ctgcagccaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacctggct ccagtacgtg attcttgatc ccgagctgga gccaggggcg ggccttgcgc
     2701 tttaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     2761 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     2821 ttaaaatttt tgatgacgtg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     2881 gggccaggat ctgcacactg gtatttcggt ttttgggccc gcggccggcg acggggcccg
     2941 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     3001 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     3061 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     3121 gccgcttccc ggccctgctc cagggggctc aaaatggagg acgcggcgct cgggagagcg
     3181 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     3241 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctgga gcttttggag
     3301 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     3361 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttggcct
     3421 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     3481 tccatttcag gtgtcgtgaa caggggatct ggagctctcg agaattctca cgcgtctgca
     3541 ggatatcaag cttgcggtac cgcgggcccg gcccaccatg accgagtaca agcccacggt
     3601 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     3661 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     3721 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     3781 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     3841 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     3901 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     3961 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4021 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     4081 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     4141 gcgcacctgg tgcatgaccc gcaagcccgg tgccggatcg ggagagggca gaggaagtct
     4201 gctaacatgc ggtgacgtcg aggagaatcc tggcccaccg gtcgccacca tgagccggct
     4261 ggacaagagc aaagtgatca acagcgccct ggaactgctg aacggcgtgg gcatcgaggg
     4321 cctgaccacc cggaagctgg cccagaaact gggcgtggaa cagcccaccc tgtactggca
     4381 cgtgaagaac aagcgggccc tgctggacgc cctgcccatc gagatgctgg accggcacca
     4441 cacccacagc tgccctctgg aaggcgagag ctggcaggac ttcctgcgga acaacgccaa
     4501 gagctacaga tgcgccctgc tgagccaccg ggacggcgcc aaagtgcacc tgggcaccag
     4561 acccaccgag aagcagtacg agacactgga aaaccagctg gccttcctgt gccagcaggg
     4621 cttcagcctg gaaaacgccc tgtacgccct gagcgccgtg ggccacttca ccctgggctg
     4681 cgtgctggaa gaacaggaac accaggtcgc caaagaggaa agagagacac ccaccaccga
     4741 cagcatgccc cccctgctga agcaggccat cgagctgttc gacagacagg gcgccgagcc
     4801 cgccttcctg ttcggcctgg aactgatcat ctgcggcctc gagaagcagc tgaagtgcga
     4861 gagcggcgga cccaccgacg ccctggacga cttcgacctg gacatgctgc ccgccgatgc
     4921 cctggatgat tttgatctgg atatgctccc tgctgacgct ctcgatgact tcgatctcga
     4981 tatgctgcca ggctaacctg caggcatgca agcttgatat caagcttatc gataatcaac
     5041 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5101 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5161 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5221 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5281 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5341 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5401 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     5461 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5521 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     5581 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cggtaaagct
     5641 gatatctcca ccccacagtg gtttactccc tatcagtgat agagaacgta tgaagagttt
     5701 actccctatc agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg
     5761 tataaggagt ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag
     5821 tgatagagaa cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt
     5881 actccctatc agtgatagag aacgtataag ctttaggcgt gtacggtggg cgcctataaa
     5941 agcagagctc gtttagtgaa ccgtcagatc gcctggagca attccacaac acttttgtct
     6001 tataccaact ttccgtacca cttcctaccc tcgtaaactt aaggttaatt aacgccacca
     6061 tggagagctc tggcaagatg gagagtggag ccggccagca gccgcagccc ccgcagccct
     6121 tcctgcctcc cgcagcctgc ttctttgcga ccgcggcggc ggcggcagcg gcggcggccg
     6181 cggcagctca gagcgcgcag cagcaacagc cgcaggcgcc gccgcagcag gcgccgcagc
     6241 tgagcccggt ggccgacagc cagccctcag ggggcggtca caagtcagcg gccaagcagg
     6301 tcaagcgcca gcgctcgtcc tctccggaac tgatgcgctg caaacgccgg ctcaacttca
     6361 gcggcttcgg ctacagcctg ccacagcagc agccggccgc cgtggcgcgc cgcaacgagc
     6421 gcgagcgcaa ccgggtcaag ttggtcaacc tgggctttgc caccctccgg gagcatgtcc
     6481 ccaacggcgc ggccaacaag aagatgagca aggtggagac gctgcgctcg gcggtcgagt
     6541 acatccgcgc gctgcagcag ctgctggacg agcacgacgc ggtgagcgct gcctttcagg
     6601 cgggcgtcct gtcgcccacc atctccccca actactccaa cgacttgaac tctatggcgg
     6661 gttctccggt ctcgtcctac tcctccgacg agggatccta cgaccctctt agcccagagg
     6721 aacaagagct gctggacttt accaactggt tctgataaat gcataccggt gcggccgcaa
     6781 tcgatcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6841 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6901 ctcatcaatg tatcttatca tgtctgcgat accgtcgacc tcgagggaat taattcgagc
     6961 tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     7021 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     7081 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7141 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7201 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7261 tctagcagca tctagaatta attccgtgta ttctatagtg tcacctaaat cgtatgtgta
     7321 tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg tacaagccta
     7381 attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa actgccgtca
     7441 gagtcggttt ggttggacga accttctgag tttctggtaa cgccgtcccg cacccggaaa
     7501 tggtcagcga accaatcagc agggtcatcg ctagccagat cctctacgcc ggacgcatcg
     7561 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     7621 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
     7681 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
     7741 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
     7801 ataagggaga gcgtcgaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     7861 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     7921 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     7981 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     8041 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     8101 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     8161 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     8221 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     8281 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     8341 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     8401 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     8461 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     8521 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     8581 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     8641 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     8701 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     8761 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     8821 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     8881 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     8941 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     9001 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     9061 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     9121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     9181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     9241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     9301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     9361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     9421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     9481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     9541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     9601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     9661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     9721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     9781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     9841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     9901 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     9961 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
    10021 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct gtggaatgtg tgtcagttag
    10081 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
    10141 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
    10201 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
    10261 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
    10321 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
    10381 gcctaggctt ttgcaaaaag cttggacaca agacaggctt gcgagatatg tttgagaata
    10441 ccactttatc ccgcgtcagg gagaggcagt gcgtaaaaag acgcggactc atgtgaaata
    10501 ctggttttta gtgcgccaga tctctataat ctcgcgcaac ctattttccc ctcgaacact
    10561 ttttaagccg tagataaaca ggctgggaca cttcacatga gcgaaaaata catcgtcacc
    10621 tgggacatgt tgcagatcca tgcacgtaaa ctcgcaagcc gactgatgcc ttctgaacaa
    10681 tggaaaggca ttattgccgt aagccgtggc ggtctgtacc gggtgcgtta ctggcgcgtg
    10741 aactgggtat tcgtcatgtc gataccgttt gtatttccag ctacgatcac gacaaccagc
    10801 gcgagcttaa agtgctgaaa cgcgcagaag gcgatggcga aggcttcatc gttattgatg
    10861 acctggtgga taccggtggt actgcggttg cgattcgtga aatgtatcca aaagcgcact
    10921 ttgtcaccat cttcgcaaaa ccggctggtc gtccgctggt tgatgactat gttgttgata
    10981 tcccgcaaga tacctggatt gaacagccgt gggatatggg cgtcgtattc gtcccgccaa
    11041 tctccggtcg ctaatctttt caacgcctgg cactgccggg cgttgttctt tttaacttca
    11101 ggcgggttac aatagtttcc agtaagtatt ctggaggctg catccatgac acaggcaaac
    11161 ctgagcgaaa ccctgttcaa accccgcttt aaacatcctg aaacctcgac gctagtccgc
    11221 cgctttaatc acggcgcaca accgcctgtg cagtcggccc ttgatggtaa aaccatccct
    11281 cactggtatc gcatgattaa ccgtctgatg tggatctggc gcggcattga cccacgcgaa
    11341 atcctcgacg tccaggcacg tattgtgatg agcgatgccg aacgtaccga cgatgattta
    11401 tacgatacgg tgattggcta ccgtggcggc aactggattt atgagtgggc cccggatctt
    11461 tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta
    11521 aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat
    11581 tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc
    11641 ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac
    11701 tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga
    11761 ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc
    11821 ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga
    11881 aaaatattct gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt
    11941 tcttactcca cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac
    12001 ctttagcttt ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac
    12061 tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
    12121 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
    12181 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
    12241 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtc
//
