pCW-eGFP

LOCUS       pCW-eGFP                8329 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Doxycycline-inducible overexpression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    pCW-eGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8329)
  AUTHORS   Fazeli PK, Zhang Y, O'Keefe J, Pesaresi T, Lun M, Lawney B, 
            Steinhauser ML
  TITLE     Prolonged fasting drives a program of metabolic inflammation in 
            human adipose tissue.
  JOURNAL   Mol Metab. 2020 Sep 28;42:101082. doi: 10.1016/j.molmet.2020.101082.
  PUBMED    32992039
REFERENCE   2  (bases 1 to 8329)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8329
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..118
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        172..486
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    180..198
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    216..234
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    251..269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    287..305
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    323..341
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    358..376
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    394..412
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     459..483
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             505..1224
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(550..571)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(811..830)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1158..1179
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        1231..1741
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1660..1678
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1751..2347
                     /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(1751..1770)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2247..2267
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2348..2401
                     /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             2402..3148
                     /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(3188..3208)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3606..3617)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3822..4055
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4127..4248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4164..4183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4218..4237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4288..4423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4350..4369
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4423..4494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4495..5355
                     /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(4713..4732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5526..6114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6015..6034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6268..6285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6402..6423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6438..6468
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6476..6492
                     /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     6481..6503
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6500..6516
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6500..6516
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6535..6555
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6537..6555
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6583..6809
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6810..6990
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7037..7162
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7655..7888
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8073..8117
                     /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 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
       61 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
      121 ggtttattac agggacagca gagatccact ttggccgcga atcgatatgt cgagtttact
      181 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
      241 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      301 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
      361 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      421 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      481 atcgcctgga gaattggcta gcccatggtg agcaagggcg aggagctgtt caccggggtg
      541 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
      601 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      661 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
      721 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      781 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      841 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      901 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      961 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     1021 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1081 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1141 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1201 ggcatggacg agctgtacaa gtaaggatcc ggggttgggg ttgcgccttt tccaaggcag
     1261 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
     1321 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
     1381 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
     1441 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
     1501 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
     1561 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
     1621 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
     1681 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
     1741 gcaattcacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     1801 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     1861 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     1921 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     1981 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     2041 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     2101 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     2161 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     2221 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     2281 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     2341 cggtgccgaa ggtagaggtt ctctcctcac ttgtggtgat gttgaagaaa accctggtcc
     2401 aatgtctaga ctggacaaga gcaaagtcat aaacggagct ctggaattac tcaatggtgt
     2461 cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac
     2521 cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa tcgagatgct
     2581 ggacaggcat catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg
     2641 gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca
     2701 tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct
     2761 gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt
     2821 tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac
     2881 acctaccacc gattctatgc ccccacttct gagacaagca attgagctgt tcgaccggca
     2941 gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca
     3001 gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact tagacatgct
     3061 cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg ctcttgacga
     3121 ttttgacctt gacatgctcc ccgggtaact aagtaaggat cgatccaaga tatcgtattc
     3181 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3241 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3301 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3361 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3421 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3481 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     3541 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3601 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3661 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3721 cgcctgtttc gcctcggcgt ccggactaga ggtaccttta agaccaatga cttacaaggc
     3781 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     3841 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
     3901 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     3961 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     4021 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
     4081 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     4141 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4201 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     4261 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4321 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4381 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctttcaaa tatgtatccg
     4441 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4681 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4801 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5041 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5101 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5341 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5641 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6181 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6241 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6301 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6361 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6421 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6481 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     6541 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     6601 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     6661 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     6721 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     6781 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     6841 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6901 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6961 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     7021 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     7081 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     7141 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     7201 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     7261 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     7321 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     7381 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     7441 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     7501 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     7561 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     7621 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     7681 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     7741 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     7801 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     7861 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     7921 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     7981 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     8041 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     8101 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     8161 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     8221 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     8281 cgaggggaca attctcgacc tcgagacaaa tggcagtatt catccacaa
//