LOCUS       sequence-451526-        9692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HSF1 fused to GFP from EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9692)
  AUTHORS   Pataki E, Gerst JE
  TITLE     Conservation of mRNA operon formation in the control of the heat 
            shock response in mammalian cells
  JOURNAL   bioRxiv 2024.10.05.616786
REFERENCE   2  (bases 1 to 9692)
  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: "bioRxiv 
            2024.10.05.616786"
COMMENT     Sequence Label: pHAGE-EF1a-HSF1-GFP
FEATURES             Location/Qualifiers
     source          1..9692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     432..451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(670..774)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     842..860
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             979..1212
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    1287..1875
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1393..1404
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1802..1822
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1895..1984)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     polyA_signal    1999..2120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2064..2083
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2124..2837,2838..2840,2841..2843))
                     /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             complement(join(2127..2837,2838..2840,2841..2843))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2169..2190)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2518..2537
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2777..2798
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2840..2849
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(2892..3263)
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     promoter        complement(4499..5677)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4508..5446
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(4531..4551)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    5782..5899
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(6192..6236)
                     /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    6421..6654
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7151..7276
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7323..7956
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8217..8233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8217..8233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8230..8252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8241..8257
                     /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(join(8265..8271,8272..8289,8290..8295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8310..8331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8571..8588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8742..9330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8822..8841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9501..9692,1..600,601..669))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
       61 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      121 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      181 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      241 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      301 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      361 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      421 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      481 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      541 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      601 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      661 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      721 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
      781 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
      841 aaataggcgt atcacgaggc cctttcgtct tcaagaatga tctagccctt tccttaatta
      901 acccgggcct ctcactctct gatattcatt tctttgcaag ttataaatac tgaataataa
      961 gatgacatga actactactg ctagagattt tccacactga ctaaaagggt ctgagggatc
     1021 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
     1081 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
     1141 ggtctaacca gagagaccca gtacaagcaa aaagcagatc ttgtcttcgt tgggagtgaa
     1201 ttagcccttc cagtcccccc ttttctttta aaaagtggct aagatctaca gctgccttgt
     1261 aagtcattgg tcttaaagga tctcaggcgg ggaggcggcc caaagggaga tccgactcgt
     1321 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     1381 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     1441 ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga
     1501 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     1561 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     1621 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     1681 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     1741 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     1801 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     1861 atccagaggt tgattaggat ctatcgattt aaacagcgta atctggaacg tcatatggat
     1921 aggatcctgc atagtccggg acgtcatagg gatagccagc gtaatctgga acatcgtatg
     1981 ggtatctaga cttgtacata agatacattg atgagtttgg acaaaccaca actagaatgc
     2041 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2101 taagctgcaa taaacaagtt gatttacttg tacagctcgt ccatgccgag agtgatcccg
     2161 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     2221 agggcggact gggtgctcag gtagtggttg tcgggcagca acacggggcc gtcgccgatg
     2281 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     2341 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     2401 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     2461 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     2521 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     2581 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     2641 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     2701 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     2761 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     2821 agctcctcgc ccttgctcac catggtggcg atggatcccg ggcccgcggt accgtcgact
     2881 gcagaattct cggagacagt ggggtccttg gctttgggag gctccgagcc tgtcagcagg
     2941 gagatggtgg ggtcctcggc gaagccgtcc ccttcggaga agtaggagcc ctctcccagc
     3001 tcaaacagca ccggcaggtc gttgctcccg gtgtccacgg agccggggtc cagcaggaac
     3061 agcggctgcg ctgtgtagtg caccagctgc ttccctgaat ccgggctgct gttctctgcc
     3121 tcgggaggcc tggggggctc ctggggagac aggagctctt ggatactggc caggctgctg
     3181 tcaaggtcag gcaggctcat gtcgggcacg gtcaccgagg ggctgaacag gtccagcagg
     3241 gcactggtgt ccacgctgaa gccgtggctg ctcagcatgg tctgcaggtt atccaggttg
     3301 gagtccatag catccaagtg gtcactgagc tcattcttgt ccaggcaggc tacgctgagg
     3361 cacttttcag gggtggaggt gggcgggggg gagggaggcc ggccctcggt gtccgtgtgg
     3421 cccctggcgt ccgtgagggc tgtgacggag gcgggggcag gttcactctc ccgcaggatg
     3481 gagtcaatga gggcggtcgg ggacaagagg gtgtccacgg aagatgggcg cccgggactc
     3541 gcctcctcta cccgggggct ctgaggcggg ctggggggct cctccttgac acgcaccagg
     3601 gggctgctgg ataggggcct ctcgtctatg ctcccgccgg gggaggccat ggggctggca
     3661 ggagccagct cggtgatgtc ggagatgatg ggtccagagc tggccacagc atcaggggcg
     3721 tagaggctgg agctgctgta ggctggggag ggggccgagt aggggcccga gccgtggacg
     3781 tgctccaggg agaactgccg gctatacttg ggcatggaat gtgctgagcc actgtcgttc
     3841 agcatcaggg ggatctttct cttcaccccc aggatccggt ttgactgcac cagtgagatc
     3901 aggaactgaa tgagcttgtt gacgactttc tgttgctggg catgcttctg ccgaaggctg
     3961 gccacctccc gccacagagc ctcattctca tgcttcatgg ccaggagctt ggagtccatg
     4021 cactcctgct tccccttcat cagctgcacg tccgtcagca gcttggtgac gctgtcctgg
     4081 cggatcttta tgtcttcact cttcagggtg gacacactgg tcactttcct cttgatgttc
     4141 tcaaggagct gctcctggcc acgcaggaag catgggtgct ggaactccgt gtcgtctctc
     4201 tctggcttga ccaggccgcc ctgctcgatg tggaccactt tccggaagcc atacatgttg
     4261 agctgccgca cgaagctggc catgttgttg tgcttgaagt acttgggcag cacctccttg
     4321 gcaaactggc cctggtcgaa cacgtggaag ctgttcccgc tcgggctcca gcagatgagc
     4381 gcgtcggtgt ccgggtcgct cacgagggtc cacagcttgg tcaggaaggc cgggacgttg
     4441 ctgggccccg ccgcgccggg gcccacgggc agatccatgg taccatgcgg ccgcgttttc
     4501 acgacacctg aaatggaaga aaaaaacttt gaaccactgt ctgaggcttg agaatgaacc
     4561 aagatccaaa ctcaaaaagg gcaaattcca aggagaatta catcaagtgc caagctggcc
     4621 taacttcagt ctccacccac tcagtgtggg gaaactccat cgcataaaac ccctcccccc
     4681 aacctaaaga cgacgtactc caaaagctgg agaactaatc gaggtgcctg gacggcgccc
     4741 ggtactccgt ggagtcacat gaagcgacgg ctgaggacgg aaaggccctt ttcctttgtg
     4801 tgggtgactc acccgcccgc tctcccgagc gccgcgtcct ccattttgtg ctccctgcag
     4861 cagggccggg aagcggccat ctttccgctc acgcaactgg tgccgaccgg gccagccttg
     4921 ccgcccaggg cggggcgata cacggcggcg cgaggccagg caccagagca ggccgggcag
     4981 cttgagacta cccccgtccg attctcggtg gccgcgctcg caggccccgc ctcgccgaac
     5041 atgtgcgctg ggacgcacgg gccccgtcgc cgcccgcggc cccaaaaacc gaaataccag
     5101 tgtgctgatc ttggcccgca tttacaagac tatcttgcca gaaaaaaagc gtcgcagcag
     5161 gtcatcaaaa attttaaatg gctagagact tatcgaaagc agcgagacag gcgcgaaggt
     5221 gccaccagat tcgcacgcgg cggccccagc gcccaggcca ggccacaact caagcacgag
     5281 gcgaaggggc tccttaagcg caaggccacg aactctccca cccacttcca acccgaagct
     5341 cgggatcaag aatcacgtac tgcagccagg tggaagtaat tcaaggcacg caagggccat
     5401 aacccgtaaa gaggccaggc ccgcgggaac cacacacggc acttacctgt gttctggcgg
     5461 caaacccgtt gcgaaaaaga acgttcacgg cgactagtgc acttatatac ggttctcccc
     5521 caccctcggg aaaaaggcgg agccagtaca cgacatcact ttcccagttt accccgcgcc
     5581 accttctcta ggcaccggtt caattgccga cccctccccc caacttctcg gggactgtgg
     5641 gcgatgtgcg ctctgcccac tgacgggcac cggagccagc gctgctcgcc gtgaggaggg
     5701 gggcgcccgc gggaggcgcc aaaacccggc gcggaggcca ctagtccaaa ctggatctct
     5761 gctgtccctg taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt
     5821 agtttgtatg tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat
     5881 cccccctttt cttttaaaat tgtggatgaa tactgccatt tgtgaattcg gcgataccgt
     5941 cgagatccgt tcactaatcg aatggatctg tctctgtctc tctctccacc ttcttcttct
     6001 attccttcgg gcctgtcggg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
     6061 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
     6121 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
     6181 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
     6241 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
     6301 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
     6361 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
     6421 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
     6481 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
     6541 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
     6601 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
     6661 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
     6721 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
     6781 cctcctccag gtctgaagat cagcggccgg ccgcttgctg tgcggtggtc ttacttttgt
     6841 tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat
     6901 gcacacaata gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct
     6961 gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt tctaacaggc
     7021 caggattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt tttaatttat
     7081 attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta attctccccc
     7141 gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta gtcaaaattt
     7201 ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag
     7261 tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc
     7321 actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
     7381 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
     7441 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
     7501 cccagtacag gcaaaaagca gctgcttata tgcaggatct gagggctcgc cactccccag
     7561 tcccgcccag gccacgcctc cctggaaagt ccccagcgga aagtcccttg tagcaagctc
     7621 gatatcagca gttcttgaag tactccggat gcagctctcg ggccacgtga tgaaatgcta
     7681 ggcggctgtc aaacctccac tctaacactt ctctctccgg gtcatccatc ccatgcaggc
     7741 tcacagggtg taacaagctg gtgttctctc ctttattggc ctcttctacc ttatctggct
     7801 caactggtac tagcttgtag caccatccaa aggtcagtgg atatctgacc cctggccctg
     7861 gtgtgtagtt ctgctaatca gggaagtagc cttgtgtgtg gtagatccac agatcaagga
     7921 tatcttgtct tctttgggag tgaattagcc cttccaacta ctaagtttgt agtacatatt
     7981 taacaaatac aatttcttta aaatgaaaat aattcagagg aatcacaggt ttagagtaaa
     8041 tgaaaccaca ggtaattggc agtggtaata gggtatgggg tgggaagttt gggatgattt
     8101 tggttagctt gagttatcca gttgatccag acatgataag atacattgat gagtttggac
     8161 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaattcgt aatcatgtca
     8221 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     8281 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     8341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc acggggagct ttttgcaaaa
     8401 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     8461 ctcggcctct gcataaataa aaaaaattag tcagccatga gcttgctgca ttaatgaatc
     8521 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     8581 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     8641 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     8701 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8761 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8821 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8881 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8941 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     9001 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     9061 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     9121 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     9181 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     9241 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     9301 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     9361 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     9421 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     9481 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     9541 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9601 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     9661 ccagatttat cagcaataaa ccagccagcc gg
//
