LOCUS       sequence-494453-        8261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for inducible expression of EGFP (all-in-one 
            Tet-On system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8261)
  AUTHORS   Heiden R, Hannig L, Kuo CJ, Ergun S, Braunger BM, Vallon M
  TITLE     WNT7A/B assemble a GPR124-RECK-LRP5/6 co-receptor complex to 
            activate beta-catenin signaling in brain endothelial cells.
  JOURNAL   J Biol Chem. 2025 Sep 4:110682. doi: 10.1016/j.jbc.2025.110682.
  PUBMED    40914247
REFERENCE   2  (bases 1 to 8261)
  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: "J Biol 
            Chem. 2025 Sep 4:110682. doi: 10.1016/j.jbc.2025.110682."
COMMENT     Sequence Label: pCW-EGFP-Blast
FEATURES             Location/Qualifiers
     source          1..8261
                     /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    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      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(549..551,552..554,555..1265)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(549..551,552..554,555..1265)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(594..615)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(855..874)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1202..1223
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1266..1331
                     /label=MCS
                     /note="multiple cloning site"
     promoter        1404..1914
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1833..1851
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1924..2319
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             2320..2373
                     /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(2374..2991,2992..2997,2998..3033,3034..3036,3037..3072,
                     3073..3075,3076..3111,3112..3120)
                     /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(3160..3180)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3578..3589)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3794..4027
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4099..4220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4136..4155)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4190..4209
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4260..4395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4395..4466
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4467..4535,4536..5327)
                     /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(4685..4704)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5498..6086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5987..6006
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6240..6257
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6374..6395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6410..6415,6416..6433,6434..6440)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6448..6464
                     /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     6453..6475
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6472..6488
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6472..6488
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6507..6527
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6509..6527
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6555..6781
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6782..6962
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7009..7134
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7627..7860
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8045..8089
                     /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 tttacttcct 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 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      601 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      661 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      721 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      781 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      841 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      901 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      961 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1021 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1081 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1141 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     1201 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1261 acaagtccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
     1321 gcccgggatc caccggatct agataactga tcataatcag ccataccacg gccggccagg
     1381 caaaaaagaa aaagtaagga tccggggttg gggttgcgcc ttttccaagg cagccctggg
     1441 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
     1501 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
     1561 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
     1621 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
     1681 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
     1741 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
     1801 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
     1861 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagcaattc
     1921 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     1981 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     2041 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     2101 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     2161 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     2221 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     2281 attcgtgaat tgctgccctc tggttatgtg tgggagggcg aaggtagagg ttctctcctc
     2341 acttgtggtg atgttgaaga aaaccctggt ccaatgtcta gactggacaa gagcaaagtc
     2401 ataaacggag ctctggaatt actcaatggt gtcggtatcg aaggcctgac gacaaggaaa
     2461 ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg
     2521 gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc
     2581 ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct
     2641 ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag
     2701 tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac
     2761 gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag
     2821 gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt
     2881 ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc
     2941 ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc
     3001 gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac
     3061 cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa
     3121 ctaagtaagg atcgatccaa gatatcgtat tcttaactat gttgctcctt ttacgctatg
     3181 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3241 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3301 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3361 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3421 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3481 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3541 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3601 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3661 gacgagtcgg atctcccttt gggccgcctc cccgcctgtt tcgcctcggc gtccggacta
     3721 gaggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     3781 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     3841 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     3901 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     3961 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     4021 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     4081 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4141 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4201 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     4261 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4321 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4381 gcttttttgg aggcctttca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4441 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     4501 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4561 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4621 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4681 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4741 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4801 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4861 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4921 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4981 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     5041 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     5101 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5161 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     5221 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     5281 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     5341 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5401 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5461 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5521 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5581 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5641 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5701 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5761 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5821 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5881 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5941 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6001 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6061 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     6121 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     6181 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     6241 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     6301 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     6361 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     6421 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     6481 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     6541 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     6601 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     6661 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     6721 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     6781 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6841 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6901 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6961 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     7021 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     7081 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     7141 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     7201 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     7261 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     7321 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     7381 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     7441 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     7501 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     7561 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     7621 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     7681 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     7741 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     7801 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     7861 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     7921 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     7981 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     8041 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     8101 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     8161 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     8221 accattatcg tttcagaccc acctcccaac cccgagggga c
//
