LOCUS       sequence-456740-        8445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible expression of RquA-FLAG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8445)
  AUTHORS   Valeros J, Jerome M, Tseyang T, Vo P, Do T, Fajardo Palomino D, 
            Grotehans N, Kunala M, Jerrett AE, Hathiramani NR, Mireku M, Magesh 
            RY, Yenilmez B, Rosen PC, Mann JL, Myers JW, Kunchok T, Manning TL, 
            Boercker LN, Carr PE, Munim MB, Lewis CA, Sabatini DM, Kelly M, Xie 
            J, Czech MP, Gao G, Shepherd JN, Walker AK, Kim H, Watson EV, 
            Spinelli JB
  TITLE     Rhodoquinone carries electrons in the mammalian electron transport 
            chain.
  JOURNAL   Cell. 2025 Jan 10:S0092-8674(24)01420-X. doi: 
            10.1016/j.cell.2024.12.007.
  PUBMED    39909039
REFERENCE   2  (bases 1 to 8445)
  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. 2025 
            Jan 10:S0092-8674(24)01420-X. doi: 10.1016/j.cell.2024.12.007."
COMMENT     Sequence Label: pcw57.1-RquA-FLAG
FEATURES             Location/Qualifiers
     source          1..8445
                     /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"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1348..1371
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1387..1897
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1816..1834
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1907..2503
                     /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(1907..1926)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2403..2423
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2504..2557
                     /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(2558..3175,3176..3181,3182..3217,3218..3220,3221..3256,
                     3257..3259,3260..3295,3296..3304)
                     /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(3344..3364)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3762..3773)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3978..4211
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4283..4404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4320..4339)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4374..4393
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4444..4579
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4579..4650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4651..4719,4720..5511)
                     /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(4869..4888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5682..6270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6171..6190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6424..6441
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6558..6579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6594..6599,6600..6617,6618..6624)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6632..6648
                     /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     6637..6659
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6656..6672
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6656..6672
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6691..6711
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6693..6711
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6739..6965
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6966..7146
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7193..7318
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7811..8044
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8229..8273
                     /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 gcattcgcca ccatgggtat gggtatgctt cggttaccca cagtctttcg ccagatgaga
      601 ccggtgtcca gggtactggc tcctcatctc actcgggctt atgccaaaat gagcggcaat
      661 cccccggagt ttcggcctgc accgtactat gatggggtcc ctgactatat ggttgatgtc
      721 tatgattggg cgtacgtcaa ccctgccaaa gctcggctgt tggatcgcaa tattgttgtt
      781 agcactctgc ttttcctgaa tgatcagcga ctgatgcggc gctatctcgc caggattaga
      841 ccgggctcaa gagtctggca ggttgcacac gtctatggtg acctggtgtc tcgagctgcc
      901 caagcggtag gcacggatgg taggtttgtc ttgacagata ttactcccgt gcaggtcgag
      961 catgcgctta aaaaactggc gccttaccca caagccgaag ttgtcagagc cgatgccagt
     1021 tcctggcaag ctgatgcgcc cttcgacctg gtgtgctctt tcttccttct gcacgaagta
     1081 ccagacgaac tcaaggcagc aatcgtcgat aatatgcttg ctcaagtcgc ccctggcggg
     1141 gagttggtgt tcgttgacta tcacagacct cgaccctggc agcccatagg ctggctgctg
     1201 aagtatgtga atagggttct cgagcctttc gcagaagcgc tttggaacaa ggatatagcg
     1261 gattacgcca cgcgcaaaga cgatttcgtg tggcaaaagg agaccgtgtt cggaggtgta
     1321 taccaaatcg tgcgagtcag agcccgggat tacaaggatg acgatgacaa gtaggactaa
     1381 ggatccgggg ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg
     1441 ctgctctggg cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt
     1501 cacgtccgtt cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac
     1561 gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg
     1621 acaaacggaa gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat
     1681 ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga
     1741 gagcagcggc cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct
     1801 gttcctgccc gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg
     1861 ctccctcgtt gaccgaatca ccgacctctc tccccagcaa ttcaccatga ccgagtacaa
     1921 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     1981 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     2041 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     2101 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     2161 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     2221 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     2281 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     2341 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     2401 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     2461 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccgaaggta gaggttctct
     2521 cctcacttgt ggtgatgttg aagaaaaccc tggtccaatg tctagactgg acaagagcaa
     2581 agtcataaac ggagctctgg aattactcaa tggtgtcggt atcgaaggcc tgacgacaag
     2641 gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa
     2701 gcgggccctg ctcgatgccc tgccaatcga gatgctggac aggcatcata cccacttctg
     2761 ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cataccgctg
     2821 tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa
     2881 acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga
     2941 gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga
     3001 acaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc
     3061 acttctgaga caagcaattg agctgttcga ccggcaggga gccgaacctg ccttcctttt
     3121 cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc
     3181 gaccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt
     3241 tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg
     3301 gtaactaagt aaggatcgat ccaagatatc gtattcttaa ctatgttgct ccttttacgc
     3361 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3421 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3481 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3541 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3601 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3661 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     3721 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3781 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3841 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tgtttcgcct cggcgtccgg
     3901 actagaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     3961 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt
     4021 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     4081 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     4141 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     4201 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     4261 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
     4321 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4381 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
     4441 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4501 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4561 ggaggctttt ttggaggcct ttcaaatatg tatccgctca tgagacaata accctgataa
     4621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4861 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5161 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5341 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5821 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6301 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6361 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6421 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6481 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6541 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6601 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6661 aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag
     6721 ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa
     6781 cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg
     6841 aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg
     6901 gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca
     6961 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     7021 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     7081 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     7141 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     7201 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     7261 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     7321 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     7381 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     7441 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     7501 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     7561 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     7621 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     7681 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     7741 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     7801 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     7861 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     7921 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     7981 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     8041 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     8101 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     8161 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     8221 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     8281 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     8341 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     8401 attcaccatt atcgtttcag acccacctcc caaccccgag gggac
//
