LOCUS       sequence-419205-        8556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of EGFP-tagged GRAM domain of GRAMD1b carrying
            a G187L mutation (pLJM1-EGFP-GRAM1b G187L).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8556)
  AUTHORS   Koh DHZ, Naito T, Na M, Yeap YJ, Rozario P, Zhong FL, Lim KL, Saheki
            Y
  TITLE     Visualization of accessible cholesterol using a GRAM domain-based 
            biosensor.
  JOURNAL   Nat Commun. 2023 Oct 25;14(1):6773. doi: 10.1038/s41467-023-42498-7.
  PUBMED    37880244
REFERENCE   2  (bases 1 to 8556)
  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: "Nat 
            Commun."; date: "2023-10-25"; volume: "14(1):6773. doi"; pages: " 
            10.1038/s41467-023-42498-7"
COMMENT     Sequence Label: pLJM1-EGFP-GRAM-H
FEATURES             Location/Qualifiers
     source          1..8556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        290..493
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     444..464
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      532..541
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(538..540,541..543,544..1254)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(538..540,541..543,544..1254)
                     /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(583..604)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(844..863)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1191..1212
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1798..1915
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1964..2474
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2393..2411
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2496..3095
                     /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(2496..2515)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2992..3012
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3223..3456
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3528..3649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3565..3584)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3619..3638
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3689..3824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3844..3863)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3845..3863)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3873..3890)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3873..3889)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3882..3904)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4031..4486
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4118..4137)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4328..4349
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4512..4616
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4617..4685,4686..5477)
                     /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(4835..4854)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5648..6236
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6137..6156
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6390..6407
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6524..6545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6560..6565,6566..6583,6584..6590)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6598..6614
                     /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     6603..6625
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6622..6638
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6622..6638
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6657..6677
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6659..6677
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6705..6931
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6932..7112
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7159..7284
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7777..8010
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8195..8239
                     /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     join(8537..8556,1)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
       61 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
      121 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgccaagta cgccccctat
      181 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      241 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      301 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      361 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      421 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      481 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccggt cgccaccatg
      541 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
      601 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      661 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      721 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
      781 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      841 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      901 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      961 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1021 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1081 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1141 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1201 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     1261 ctcagatctc gagaaaagca gagaaatgaa gacttcagaa agctctttaa gcagcttcca
     1321 gacacggagc gcctcattgt tgattactca tgtgcactcc aaagagacat tctccttcag
     1381 ggccgactct acctctctga aaattggatc tgcttctaca gcaacatctt ccgctgggaa
     1441 actctgctga cagtccgttt gaaagacatc tgttccatga ctaaagaaaa aacagctcgc
     1501 ctcattccca atgccatcca agtttgcact gattcagaaa agcacttctt cacttcgttt
     1561 ctcgcccggg ataggacata tatgatgatg ttccggctct ggcagaatgc tctccttgaa
     1621 aagtgaggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     1681 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1741 aaacataaaa tgaatgcaat tctcgacctc gagacaaatg gcagtattca tccacaattt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     1921 ttattacagg gacagcagag atccactttg gccgcggctc gagggggttg gggttgcgcc
     1981 ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt ggttccggga
     2041 aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc agcgtcaccc
     2101 ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg cccctaagtc
     2161 gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca
     2221 ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga ccgcgatggg
     2281 ctgtggccaa tagcggctgc tcagcagggc gcgccgagag cagcggccgg gaaggggcgg
     2341 tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg
     2401 cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg
     2461 acctctctcc ccagggggat ccaccggagc ttaccatgac cgagtacaag cccacggtgc
     2521 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     2581 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     2641 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     2701 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     2761 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     2821 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     2881 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     2941 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     3001 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     3061 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     3121 cgaccgaaag gagcgcacga ccccatgcat cggtaccttt aagaccaatg acttacaagg
     3181 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     3241 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     3301 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     3361 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     3421 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     3481 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     3541 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3601 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     3661 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3721 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3781 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     3841 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     3901 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     3961 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     4021 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4081 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4141 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4201 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4261 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4321 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4381 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     4441 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     4501 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4561 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     4621 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4681 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4741 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4801 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4861 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4921 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4981 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5041 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5101 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5161 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5221 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5281 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5341 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5401 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5461 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5521 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5581 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5641 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5701 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5761 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5821 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5881 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5941 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6001 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6061 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6121 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6181 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6241 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6301 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6361 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     6421 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     6481 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     6541 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     6601 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     6661 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat
     6721 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa
     6781 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
     6841 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
     6901 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
     6961 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     7021 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     7081 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
     7141 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
     7201 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     7261 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
     7321 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
     7381 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
     7441 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
     7501 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
     7561 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
     7621 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     7681 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     7741 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     7801 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     7861 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     7921 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     7981 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     8041 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     8101 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     8161 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     8221 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     8281 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     8341 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     8401 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     8461 cagatccatt cgattagtga acggatctcg acggtatcga tcacgagact agcctcgagc
     8521 ggccgccccc ttcaccgagg gcctatttcc catgat
//
