LOCUS       sequence-440957-        8723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP gene expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8723)
  AUTHORS   Ali RH, Orellana EA, Lee SH, Chae YC, Chen Y, Clauwaert J, Kennedy 
            AL, Gutierrez AE, Papke DJ, Valenzuela M, Silverman B, Falzetta A, 
            Ficarro SB, Marto JA, Fletcher CDM, Perez-Atayde A, Alcindor T, 
            Shimamura A, Prensner JR, Gregory RI, Gutierrez A
  TITLE     A methyltransferase-independent role for METTL1 in tRNA 
            aminoacylation and oncogenic transformation.
  JOURNAL   Mol Cell. 2025 Mar 6;85(5):948-961.e11. doi: 
            10.1016/j.molcel.2025.01.003. Epub 2025 Jan 31.
  PUBMED    39892392
REFERENCE   2  (bases 1 to 8723)
  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"; doi: 
            "10.1016/j.molcel.2025.01.003"; journalName: "Mol Cell"; date: 
            "2025-03-6- 6"; volume: "85"; issue: "5"; pages: "948-961.e11"
COMMENT     Sequence Label: pdest N-FLAG EGFP puro
FEATURES             Location/Qualifiers
     source          1..8723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    229..250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(265..270,271..288,289..295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    303..319
                     /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     308..330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     327..343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     327..343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     362..382
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        364..382
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        410..636
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             637..817
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    864..989
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1482..1715
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1900..1944
                     /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"
     misc_feature    2211..2327
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2350..2653
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2654..2857
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2808..2828
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2854..2878
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2962..2986
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2995..3018
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(3019..3021,3022..3735)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3019..3021,3022..3732)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3061..3082)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3322..3341)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3669..3690
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3737..3761)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3792..4380
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3845..3865)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4263..4274)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4394..4893
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4429..4452)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4826..4845
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4914..5513
                     /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(4914..4933)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5410..5430
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5651..5884
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5956..6077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5993..6012)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6047..6066
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6117..6252
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6272..6291)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6273..6291)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6301..6318)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6301..6317)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6310..6332)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6459..6914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6546..6565)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6756..6777
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6940..7044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7045..7113,7114..7905)
                     /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(7263..7282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8076..8664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8565..8584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      181 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      241 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      301 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
      361 cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat
      421 gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga
      481 gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta
      541 ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga
      601 gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc
      661 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      721 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      781 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      841 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      901 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      961 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     1021 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     1081 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     1141 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     1201 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     1261 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     1321 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1381 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1441 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1501 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1561 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1621 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1681 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1741 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1801 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1861 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1921 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1981 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     2041 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     2101 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     2161 agagacagat ccattcgatt agtgaacgga tctcgacggt atcggttaac ttttaaaaga
     2221 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     2281 atacaaacta aagaattaca aaaacaaatt acaaaattca aaattttatc gataagcttg
     2341 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     2401 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     2461 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     2521 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     2581 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     2641 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     2701 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     2761 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     2821 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     2881 ctggagacgc catccacgct gttttgacct ccatagaaga caccgactct agtccagtgt
     2941 ggtggaattc tgcagatatc aacaagtttg tacaaaaaag caggcttcac catggactac
     3001 aaagacgatg acgacaaggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3061 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3121 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3181 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3241 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3301 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3361 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3421 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3481 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     3541 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3601 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     3661 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3721 gagctgtaca agtgacaccc agctttcttg tacaaagtgg ttgatatcca gcacagtggc
     3781 ggccgctcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     3841 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     3901 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     3961 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4021 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4081 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4141 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     4201 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4261 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4321 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     4381 ctggaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta
     4441 gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca
     4501 tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact
     4561 cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa
     4621 atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag
     4681 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
     4741 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
     4801 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
     4861 ctgttctcct cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg
     4921 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     4981 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     5041 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5101 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5161 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     5221 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     5281 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5341 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5401 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5461 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     5521 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatct cgagggcccg
     5581 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5641 aaggggggac tggaagggct agctcactcc caacgaagac aagatctgct ttttgcttgt
     5701 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5761 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5821 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5881 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     5941 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6001 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6061 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     6121 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6181 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6241 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     6301 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     6361 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     6421 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     6481 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     6541 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     6601 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6661 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     6721 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     6781 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     6841 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     6901 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6961 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     7021 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     7081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     7141 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     7201 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     7261 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     7321 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     7381 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7441 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     7501 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7561 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     7621 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     7681 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7741 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7801 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7861 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     8041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     8101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     8161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     8221 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     8281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8701 ttttgctggc cttttgctca cat
//
