LOCUS       sequence-431942-        9059 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Proximity biotinylation control of OSBP(PH domain).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9059)
  AUTHORS   Hollingsworth LR, Veeraraghavan P, Paulo JA, Harper JW
  TITLE     Spatiotemporal proteomic profiling of cellular responses to NLRP3 
            agonists
  JOURNAL   bioRxiv 2024.04.19.590338
REFERENCE   2  (bases 1 to 9059)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.04.19.590338"
COMMENT     Sequence Label: pLV UBC OSBP(PH)-APEX2-3xFLAG-IRES-eGFP
FEATURES             Location/Qualifiers
     source          1..9059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    217..333
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        336..734
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      1143..1152
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1149..1898
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="MGKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAG
                     TFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVA
                     VEVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGH
                     TIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVD
                     KYAADEDAFFADYAEAHQKLSELGFADA"
     CDS             1917..1982
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2019..2571
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2163..2180)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2390..2409
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2583..2585,2586..2588,2589..3302)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2583..2585,2586..2588,2589..3299)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2628..2649)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2889..2908)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3236..3257
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3318..3906
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3371..3391)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3789..3800)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3993..4226
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4298..4419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4335..4354)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4389..4408
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4459..4594
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4614..4633)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4615..4633)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4643..4660)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4643..4659)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4652..4674)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4801..5256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4888..4907)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5098..5119
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5282..5386
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5387..5455,5456..6247)
                     /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(5605..5624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6418..7006
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6907..6926
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7160..7177
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7294..7315
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7330..7335,7336..7353,7354..7360)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7368..7384
                     /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     7373..7395
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7392..7408
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7392..7408
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7427..7447
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7429..7447
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7475..7701
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7702..7882
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7929..8054
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8547..8780
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8965..9009
                     /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 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
       61 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
      121 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
      181 tcgattagtg aacggatctc gacggtatcg gttaactttt aaaagaaaag gggggattgg
      241 ggggtatagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
      301 attacaaaaa caaattacaa aattcaaaat tttatggcct ccgcgccggg ttttggcgcc
      361 tcccgcgggc gcccccctcc tcacggcgag cgctgccacg tcagacgaag ggcgcagcga
      421 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
      481 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
      541 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
      601 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
      661 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
      721 tgatcgtcac ttggggagac gccatccacg ctgttttgac ctccatagaa gacaccgacg
      781 ccaccatgtc aggatcggct cgagagggct ggctcttcaa atggaccaat tatatcaaag
      841 gctaccagcg gcgatggttc gtgctgagca acgggctcct gagctactac agatcaaagg
      901 cagaaatgag acatacctgc cgtggtacca tcaacctcgc cacagccaac atcaccgtgg
      961 aggactcctg caacttcatc atttccaatg ggggtgctca gacctaccat ctgaaagcta
     1021 gttcagaagt tgagcggcag cgctgggtga cggccctgga actggccaag gccaaagctg
     1081 tgaagggcgg atccggggga agcggcggat ccatcgaaga cggaagagcg gatccaccgg
     1141 tcgccaccat gggaaagtct tacccaactg tgagtgctga ttaccaggac gccgttgaga
     1201 aggcgaagaa gaagctcaga ggcttcatcg ctgagaagag atgcgctcct ctaatgctcc
     1261 gtttggcatt ccactctgct ggaacctttg acaagggcac gaagaccggt ggacccttcg
     1321 gaaccatcaa gcaccctgcc gaactggctc acagcgctaa caacggtctt gacatcgctg
     1381 ttaggctttt ggagccactc aaggcggagt tccctatttt gagctacgcc gatttctacc
     1441 agttggctgg cgttgttgcc gttgaggtca cgggtggacc taaggttcca ttccaccctg
     1501 gaagagagga caagcctgag ccaccaccag agggtcgctt gcccgatccc actaagggtt
     1561 ctgaccattt gagagatgtg tttggcaaag ctatggggct tactgaccaa gatatcgttg
     1621 ctctatctgg gggtcacact attggagctg cacacaagga gcgttctgga tttgagggtc
     1681 cctggacctc taatcctctt attttcgaca actcatactt cacggagttg ttgagtggtg
     1741 agaaggaagg tctccttcag ctaccttctg acaaggctct tttgtctgac cctgtattcc
     1801 gccctctcgt tgacaaatat gcagcggacg aagatgcctt ctttgctgat tacgctgagg
     1861 ctcaccaaaa gctttccgag cttgggtttg ctgatgccgg cggatccggg ggaagcgact
     1921 acaaggacca cgacggtgac tacaaggacc acgacatcga ctacaaggac gacgacgaca
     1981 agtaatgagt cgacattaat taacgccccc cccccctaac gttactggcc gaagccgctt
     2041 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     2101 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     2161 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagtag ttcctctgga
     2221 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     2281 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     2341 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     2401 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     2461 tctggggcct cggtgcacat gctttacatg tatttagtcg aggttaaaaa aacgtctagg
     2521 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
     2581 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2641 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2701 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2761 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     2821 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2881 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2941 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3001 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3061 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3121 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3181 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3241 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3301 aaagcggccg cgtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3361 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3421 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3481 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3541 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3601 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3661 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     3721 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     3781 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3841 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3901 ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga
     3961 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     4021 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     4081 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     4141 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     4201 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     4261 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     4321 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4381 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
     4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4561 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
     4621 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4681 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4741 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     4801 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4861 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4921 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4981 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     5041 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     5101 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     5161 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     5221 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     5281 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5341 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5401 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5461 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5521 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5581 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5641 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5701 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5761 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5821 gctaaccgct tttttgcaca acatgggaga tcatgtaact cgccttgatc gttgggaacc
     5881 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5941 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6001 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6061 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6121 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6181 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6241 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6301 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6361 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6421 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6481 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6541 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6601 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6661 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6721 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6781 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6841 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6901 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6961 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7021 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7081 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7141 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7201 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     7261 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     7321 accccaggct ttccacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     7381 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
     7441 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
     7501 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
     7561 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
     7621 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
     7681 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
     7741 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     7801 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7861 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
     7921 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
     7981 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
     8041 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     8101 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     8161 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     8221 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     8281 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     8341 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     8401 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     8461 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     8521 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     8581 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     8641 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     8701 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     8761 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     8821 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     8881 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     8941 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     9001 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcat
//
