LOCUS       sequence-210611-        8996 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Tunable system for controlled gene expression and protein stability 
            in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8996)
  AUTHORS   Pedone E, Postiglione L, Aulicino F, Rocca DL, Montes-Olivas S, 
            Khazim M, di Bernardo D, Pia Cosma M, Marucci L
  TITLE     A tunable dual-input system for on-demand dynamic gene expression 
            regulation.
  JOURNAL   Nat Commun. 2019 Oct 2;10(1):4481. doi: 10.1038/s41467-019-12329-9.
  PUBMED    31578371
REFERENCE   2  (bases 1 to 8996)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-10-2"; volume: "10(1):4481. doi"; pages: " 
            10.1038/s41467-019-12329-9"
COMMENT     Sequence Label: pLVX_TRE3G_DD::mCherry
FEATURES             Location/Qualifiers
     source          1..8996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..350
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    81..99
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    117..135
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    153..171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    189..207
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    225..243
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     289..313
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             871..1578
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1013..1031)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1281..1300
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        1624..2123
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1659..1682)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2056..2075
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2144..2743
                     /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(2144..2163)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2640..2660
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2760..3348
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2813..2833)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3231..3242)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3555..4188
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4316..4332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4316..4332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4329..4351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4340..4356
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4364..4370,4371..4388,4389..4394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4409..4430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4547..4564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4718..5306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4798..4817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5477..6268,6269..6337))
                     /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     6100..6119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6338..6442)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6490..6624
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6527..6546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6581..6600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6792..7425
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7472..7597
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8094..8327
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8512..8556
                     /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    8819..8934
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
ORIGIN
        1 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag
       61 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc
      121 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta
      181 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag
      241 agaacgtatg tcgaggtagg cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt
      301 gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatactt ggatccgggc
      361 ccgcggccgc cccccgcgga tcccccgggc cacatgatca gcctgatcgc cgccctggcc
      421 gtggactacg tgatcggcat ggagaacgcc atgccctgga acctgcccgc cgacctggcc
      481 tggttcaagc gcaacaccct gaacaagccc gtgatcatgg gccgccacac ctgggagagc
      541 atcggccgcc ccctgcccgg ccgcaagaac atcatcctga gcagccagcc cggcaccgac
      601 gaccgcgtga cctgggtgaa gagcgtggac gaggccatcg ccgcctgcgg cgacgtgccc
      661 gagatcatgg tgatcggcgg cggccgcgtg atcgagcagt tcctgcccaa ggcccagaag
      721 ctgtacctga cccacatcga cgccgaggtg gagggcgaca cccacttccc cgactacgag
      781 cccgacgact gggagagcgt gttcagcgag ttccacgacg ccgacgccca gaacagccac
      841 agctactgct tcgagatcct ggagcgccgc gtgagcaagg gcgaggagga taacatggcc
      901 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
      961 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1021 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     1081 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     1141 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     1201 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1261 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1321 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     1381 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     1441 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     1501 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     1561 gacgagctgt acaagtaacc cggggaattc ctcgagaagg gacgcgtcat atggaattct
     1621 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     1681 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     1741 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     1801 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     1861 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     1921 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     1981 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     2041 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     2101 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc
     2161 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     2221 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     2281 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     2341 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     2401 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     2461 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     2521 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     2581 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     2641 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     2701 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgggcgc gtctggaaca
     2761 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2821 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2881 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2941 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3001 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3061 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3121 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     3181 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3241 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3301 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt
     3361 ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     3421 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     3481 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     3541 agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     3601 gtggatctac cacacacaag gctacttccc tgattagcag aactacacac cagggccagg
     3661 ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa
     3721 ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg
     3781 gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca
     3841 tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta
     3901 caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg
     3961 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
     4021 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     4081 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     4141 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     4201 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     4261 cttgacattg ctagcgttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4321 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6481 cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6541 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6601 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
     6661 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
     6721 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
     6781 aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg
     6841 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt
     6901 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt
     6961 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat
     7021 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca
     7081 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa
     7141 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg
     7201 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag
     7261 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     7321 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     7381 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     7441 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     7501 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     7561 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     7621 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     7681 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     7741 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     7801 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     7861 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     7921 caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     7981 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     8041 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     8101 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     8161 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     8221 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     8281 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     8341 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     8401 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     8461 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     8521 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     8581 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     8641 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     8701 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     8761 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgcctt
     8821 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     8881 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     8941 ttattacagg gacagcagag atccagttta tcgatgaggc cctttcgtct tcactc
//
