LOCUS       sequence-272431-       10560 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Flag and GFP-tagged OSBP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10560)
  AUTHORS   Lim CY, Davis OB, Shin HR, Zhang J, Berdan CA, Jiang X, Counihan JL,
            Ory DS, Nomura DK, Zoncu R
  TITLE     ER-lysosome contacts enable cholesterol sensing by mTORC1 and drive 
            aberrant growth signalling in Niemann-Pick type C.
  JOURNAL   Nat Cell Biol. 2019 Oct;21(10):1206-1218. doi: 
            10.1038/s41556-019-0391-5. Epub 2019 Sep 23.
  PUBMED    31548609
REFERENCE   2  (bases 1 to 10560)
  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"; doi: 
            "10.1038/s41556-019-0391-5"; journalName: "Nat Cell Biol"; date: 
            "2019-10"; volume: "21"; issue: "10"; pages: "1206-1218"
COMMENT     Sequence Label: pLJM1-FLAG-GFP-OSBP
FEATURES             Location/Qualifiers
     source          1..10560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..172
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2727..2844
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2893..3403
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3322..3340
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3425..4024
                     /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(3425..3444)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3921..3941
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4152..4385
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4457..4578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4494..4513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4548..4567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4618..4753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4773..4792)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4774..4792)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4802..4819)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4802..4818)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4811..4833)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4960..5415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5047..5066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5257..5278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5441..5545
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5546..5614,5615..6406)
                     /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(5764..5783)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6577..7165
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7066..7085
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7319..7336
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7453..7474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7489..7494,7495..7512,7513..7519)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7527..7543
                     /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     7532..7554
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7551..7567
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7551..7567
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7586..7606
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7588..7606
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7634..7860
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7861..8041
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8088..8213
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8706..8939
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9124..9168
                     /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"
     primer_bind     9466..9486
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        9775..9978
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9929..9949
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             10022..10045
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(10061..10063,10064..10560,1..214)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(10061..10063,10064..10560,1..214)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(10103..10124)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(10364..10383)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
       61 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
      121 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      181 cgggatcact ctcggcatgg acgagctgta caagggaggc ggtggatcgg gcgggggtgg
      241 atcaaagtcg acgatggctg ctacggagct gagaggagtg gtgggtccag gaccagcagc
      301 tattgcagca cttggaggtg gaggagccgg tcctccagtt gtaggaggag gtggaggacg
      361 cggagatgcg ggaccaggct ccggtgccgc gtcagggacg gtggtcgcag cagcagctgg
      421 aggcccggga cctggtgcag gtggagttgc agctgcaggt cctgcacctg ctccgcctac
      481 tggaggttca ggaggttcag gagctggagg ttcaggatcg gctcgagagg gctggctctt
      541 caaatggacc aattatatca aaggctacca gcggcgatgg ttcgtgctga gcaacgggct
      601 cctgagctac tacagatcaa aggcagaaat gagacatacc tgccgtggta ccatcaacct
      661 cgccacagcc aacatcaccg tggaggactc ctgcaacttc atcatttcca atgggggtgc
      721 tcagacctac catctgaaag ctagttcaga agttgagcgg cagcgctggg tgacggccct
      781 ggaactggcc aaggccaaag ctgtgaagat gctggcagag tcagatgaat caggagatga
      841 agagtctgtc tcacaaactg acaagactga gctgcagaat acccttcgga ccctctctag
      901 caaagtagag gacttgagca cgtgcaatga cttgatagct aagcatggca cagctctgca
      961 gcgttctctc agtgagctgg agtccctgaa gttgcctgct gagagcaatg aaaagatcaa
     1021 acaggtcaac gaacgagcca cactctttag gataacatcc aatgccatga tcaacgcctg
     1081 cagagatttc ctcatgttag cccagaccca tagtaaaaaa tggcaaaagt cactacagta
     1141 tgaaagagac cagcgtatcc gactggaaga aaccctcgag cagctggcga agcagcataa
     1201 tcacctggag agggccttcc gaggagccac ggtgctgccg gcaaacactc ctggcaatgt
     1261 gggttctggt aaagatcagt gctgctctgg caaaggggac atgagcgatg aagatgatga
     1321 gaatgaattt tttgatgcac ctgagatcat caccatgcct gaaaatttgg gccacaaacg
     1381 tactggcagc aatatcagtg gagccagcag tgacatcagc cttgatgaac agtacaagca
     1441 tcagctggag gagaccaaaa aggaaaagag aaccagaata ccatacaagc caaactatag
     1501 cctcaattta tggagcatca tgaagaactg cattggaaaa gaactctcta agatccccat
     1561 gccggtaaac tttaatgagc ccttgtccat gcttcagcgc cttactgaag atctggaata
     1621 ccatgagctg ttagaccgag ctgcaaaatg tgagaattct ctagaacagc tctgttatgt
     1681 tgcagctttc accgtgtcct cctactccac tactgtcttc cgcaccagta agccattcaa
     1741 cccactgctt ggggagacct ttgagctgga ccgattagag gagaatgggt accgatccct
     1801 ctgtgaacag gtgagtcatc atccccctgc tgctgcgcac catgctgagt ccaaaaatgg
     1861 ctggacattg cgtcaggaaa tcaaaatcac cagcaagttt cgaggcaaat acctctccat
     1921 tatgcccctc ggtaccattc attgtatttt ccatgcaact gggcaccact acacttggaa
     1981 gaaagttacc acaactgtac acaacattat tgtgggcaag ttgtggatag atcagtctgg
     2041 cgaaattgat attgtgaatc acaagacagg agacaagtgt aatcttaaat ttgttcctta
     2101 tagctacttc tctcgggatg tagcaagaaa ggtgacgggg gaagtgacag atccatcagg
     2161 aaaagtccac tttgctcttc tggggacgtg ggatgagaaa atggaatgtt tcaaagtaca
     2221 gccagtcatt ggggaaaatg ggggtgatgc tcgacagaga ggccatgaag cagaggaaag
     2281 cagggtcatg ctgtggaaaa ggaatccttt accgaagaat gcagaaaaca tgtactactt
     2341 ctcagagctt gctctgactc tcaatgcttg ggaaagtggc actgccccca cagacagccg
     2401 gttacgacct gaccagagac tgatggaaaa tggacgctgg gatgaagcaa atgcggagaa
     2461 gcagcgcctg gaggaaaaac aaagactttc cagaaagaag agagaagcgg aagctatgaa
     2521 agccacagag gatggcacac catatgatcc ctataaggca ctgtggtttg agcggaagaa
     2581 ggaccctgtt accaaggagt taacccatat ttatagggga gaatactggg agtgtaaaga
     2641 aaaacaggac tggagctcat gcccggacat tttctgagcg cggatcaatt ctcgacctcg
     2701 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2761 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2821 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
     2881 ccgcggctcg agggggttgg ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg
     2941 acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg ggtctcgcac
     3001 attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg tgggcccccc
     3061 ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg
     3121 gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac agcgccaggg
     3181 agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct cagcagggcg
     3241 cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg
     3301 ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc gcacgtcggc
     3361 agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc caccggagct
     3421 taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     3481 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     3541 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     3601 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     3661 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     3721 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     3781 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     3841 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     3901 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     3961 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     4021 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc
     4081 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     4141 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     4201 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     4261 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     4321 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     4381 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     4441 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4501 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4561 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     4621 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4681 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4741 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     4801 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     4861 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     4921 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     4981 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5041 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5101 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5161 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5221 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5281 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5341 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5401 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5461 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5521 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5581 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5641 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5701 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5761 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5821 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5881 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5941 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6001 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6061 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6121 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6181 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6241 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6301 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6361 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6421 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6481 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6541 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6601 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6661 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6721 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6781 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6841 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6901 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6961 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7021 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7081 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7141 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7201 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7261 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7321 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7381 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     7441 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     7501 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     7561 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     7621 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg
     7681 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
     7741 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga
     7801 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac
     7861 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7921 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7981 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     8041 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     8101 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     8161 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     8221 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     8281 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     8341 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     8401 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     8461 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     8521 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     8581 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     8641 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     8701 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     8761 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     8821 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     8881 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     8941 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     9001 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     9061 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     9121 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     9181 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     9241 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     9301 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     9361 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     9421 cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg cctatttccc
     9481 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     9541 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     9601 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgcc aagtacgccc
     9661 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     9721 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     9781 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     9841 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     9901 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     9961 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtgccat
    10021 ggactacaag gatgacgatg acaagggcgg aggtggatct gtgagcaagg gcgaggagct
    10081 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
    10141 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
    10201 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
    10261 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
    10321 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
    10381 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
    10441 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
    10501 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
//
