pLV-EF1a-MinE(L4E)-EGFP_IRES-Blast

LOCUS       pLV-EF1a-MinE(L4E)-EGFP_IRES-Blast                9544 bp ds-DNA     circular SYN 26-JAN-2024
DEFINITION  Expression of a faster-running MinE mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pLV-EF1a-MinE(L4E)-EGFP_IRES-Blast
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9544)
  AUTHORS   Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM
  TITLE     A programmable reaction-diffusion system for spatiotemporal cell 
            signaling circuit design.
  JOURNAL   Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi: 
            10.1016/j.cell.2023.12.007.
  PUBMED    38181787
REFERENCE   2  (bases 1 to 9544)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 26, 2024 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9544
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             34..667
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(795..811)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(795..811)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(808..830)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    819..835
                     /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)."
     promoter        complement(843..873)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    888..909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1026..1043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1197..1785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1277..1296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1956..2816)
                     /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     2579..2598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2817..2921)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2969..3103
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3006..3025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3060..3079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3271..3904
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3951..4076
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4573..4806
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4991..5035
                     /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    5298..5413
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        5491..5702
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          5703..6641
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6598..6618
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      6657..6666
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6960..7679
                     /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             6960..7676
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7005..7026)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7266..7285)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7613..7634
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7923..7940)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8150..8169
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    8783..9371
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8836..8856)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9254..9265)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
       61 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
      121 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
      181 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
      241 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
      301 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
      361 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
      421 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
      481 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      541 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
      601 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
      661 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
      721 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc
      781 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
      841 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
      901 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
      961 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1621 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     2101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     2161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     2221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     2281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     2341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     2401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     2461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     2521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     2581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     2641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     2701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     2761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     2821 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     2881 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     2941 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa
     3001 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3061 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc
     3121 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg
     3181 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat
     3241 ttgttaaata tgtactacaa acttagtagt tggaagggct aattcactcc caaagaagac
     3301 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact
     3361 acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac
     3421 cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc
     3481 ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca
     3541 gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct
     3601 gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg
     3661 gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt
     3721 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     3781 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     3841 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     3901 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     3961 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     4021 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     4081 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     4141 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     4201 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     4261 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     4321 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     4381 agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg
     4441 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     4501 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     4561 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     4621 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     4681 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     4741 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     4801 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     4861 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     4921 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     4981 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     5041 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     5101 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     5161 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
     5221 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
     5281 atctcgacgg tatcgccttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     5341 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     5401 aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttat cgatgaggcc
     5461 ctttcgtctt cactcgaggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     5521 agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
     5581 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     5641 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     5701 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
     5761 tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg
     5821 gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag
     5881 ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct
     5941 gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc
     6001 tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg
     6061 tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc
     6121 ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg
     6181 ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc
     6241 ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct
     6301 caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa
     6361 gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca
     6421 ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt
     6481 tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc
     6541 acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca
     6601 agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggatccgcca
     6661 ccatggccct tgaggacttc ttcctgtctc gaaaaaagaa cacagcgaat atcgcgaaag
     6721 aacggttgca aattatagtg gcggagagac gacgaagcga cgcggagcct cattaccttc
     6781 ctcaattgcg aaaagacata ctggaagtga tctgcaagta cgttcaaatt gaccctgaga
     6841 tggtgaccgt gcaacttgaa cagaaggatg gagatataag catcctggaa ttgaacgtaa
     6901 cccttcccga ggcggaagaa cttaaagaca gcgccggaag tgctggctct gcgggtacca
     6961 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     7021 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     7081 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     7141 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     7201 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     7261 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     7321 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     7381 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     7441 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     7501 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     7561 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     7621 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag
     7681 cggccgctct agagtcgacg ggccgcggta acaattgtta actaacttaa gctagcaacg
     7741 gtttccctct agcgggatca attccgcccc cccccctaac gttactggcc gaagccgctt
     7801 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
     7861 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
     7921 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
     7981 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
     8041 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
     8101 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
     8161 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
     8221 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
     8281 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgataatacc atggtcatga
     8341 aaacatttaa catttctcaa caagatctag aattagtaga agtagcgaca gagaagatta
     8401 caatgcttta tgaggataat aaacatcatg tgggagcggc aattcgtacg aaaacaggag
     8461 aaatcatttc ggcagtacat attgaagcgt atataggacg agtaactgtt tgtgcagaag
     8521 ccattgcgat tggtagtgca gtttcgaatg gacaaaagga ttttgacacg attgtagctg
     8581 ttagacaccc ttattctgac gaagtagata gaagtattcg agtggtaagt ccttgtggta
     8641 tgtgtaggga gttgatttca gactatgcac cagattgttt tgtgttaata gaaatgaatg
     8701 gcaagttagt caaaactacg attgaagaac tcattccact caaatatacc cgaaattaac
     8761 ccgggcgggg cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac
     8821 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     8881 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     8941 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     9001 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     9061 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     9121 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     9181 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     9241 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     9301 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     9361 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca
     9421 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     9481 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     9541 gtag
//