LOCUS       sequence-358805-        9838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SED1 biosensor for stable integration and expression in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9838)
  AUTHORS   Cuevas-Velazquez CL, Vellosillo T, Guadalupe K, Schmidt HB, Yu F, 
            Moses D, Brophy JAN, Cosio-Acosta D, Das A, Wang L, Jones AM, 
            Covarrubias AA, Sukenik S, Dinneny JR
  TITLE     Intrinsically disordered protein biosensor tracks the 
            physical-chemical effects of osmotic stress on cells.
  JOURNAL   Nat Commun. 2021 Sep 14;12(1):5438. doi: 10.1038/s41467-021-25736-8.
  PUBMED    34521831
REFERENCE   2  (bases 1 to 9838)
  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: "2021-09-14"; volume: "12(1):5438. doi"; pages: " 
            10.1038/s41467-021-25736-8"
COMMENT     Sequence Label: pHBS1449 SED1-Lenti
FEATURES             Location/Qualifiers
     source          1..9838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     276..293
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    410..431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(446..451,452..469,470..476)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    484..500
                     /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     489..511
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     508..524
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     508..524
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     543..563
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        545..563
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        591..817
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             818..998
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1045..1170
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1663..1896
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2081..2125
                     /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    2392..2508
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2531..2834
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2835..3038
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2989..3009
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3035..3059
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4653..4664
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    5088..5676
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5141..5161)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5559..5570)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        5690..6189
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5725..5748)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6122..6141
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6210..6809
                     /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(6210..6229)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6706..6726
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             6947..7180
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7252..7373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7289..7308)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7343..7362
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7413..7548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7568..7587)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7569..7587)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7597..7614)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7597..7613)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7606..7628)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7755..8210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7842..7861)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8052..8073
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8236..8340
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8341..8409,8410..9201)
                     /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(8559..8578)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(9372..9838,1..122)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
       61 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      121 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      181 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      241 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      301 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      361 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      421 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      481 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      541 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
      601 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
      661 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
      721 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
      781 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
      841 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
      901 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
      961 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     1021 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     1081 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     1141 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     1201 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     1261 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     1321 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     1381 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     1441 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     1501 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1561 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1621 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1681 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1741 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1801 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1861 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1921 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1981 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     2041 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     2101 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     2161 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     2221 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     2281 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     2341 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa cttttaaaag
     2401 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2461 catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat cgataagctt
     2521 gggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     2581 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     2641 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     2701 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     2761 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     2821 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     2881 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     2941 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     3001 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
     3061 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgactc tagtccagtg
     3121 tggtggaatt ctgcagatat ggtgagcaaa ggtgaagaac tgtttaccgg tgttgttccg
     3181 attctggttg aactggatgg tgacgttaat ggtcacaaat tttcagttag cggtgaaggc
     3241 gaaggtgatg caacctatgg taaactgacc ctgaaattta tctgtaccac cggcaaactg
     3301 ccggttccgt ggccgaccct ggttaccacc ctgagctggg gtgttcagtg ttttgcacgt
     3361 tatccggatc acatgaaaca gcacgatttt ttcaaaagcg caatgccgga aggttatgtt
     3421 caagaacgta ccatcttctt caaagatgac ggcaactata aaacccgtgc cgaagttaaa
     3481 tttgaaggtg ataccctggt gaatcgcatt gaactgaaag gcatcgattt taaagaggat
     3541 ggtaatatcc tgggccacaa actggaatat aatgccattc atggcaacgt gtatatcacc
     3601 gcagataaac agaaaaacgg catcaaagca aattttggcc tgaactgcaa tattgaagat
     3661 ggttcagttc agctggcaga tcattatcag cagaataccc cgattggtga tggtccggtt
     3721 ctgctgccgg ataatcatta tctgagcacc cagagcaaac tgagcaaaga tccgaatgaa
     3781 aaacgtgatc acatggtgct gctggaattt gttaccgcag caggtattac cctgggtatg
     3841 gatgaactgt ataaagagct catgcagtcg atgaaagaaa cagcttcgaa tattgcagct
     3901 tctgcaaaat ctggcatgga caaaaccaaa gctaccttgg aggaaaaggc ggagaagatg
     3961 aagacacgag accctgttca gaaacagatg gctacacagg ttaaagaaga taagatcaat
     4021 caagctgaga tgcagaagag agaaacgcgt cagcacaacg cggccatgaa agaagcggct
     4081 ggagccggaa ccggtttagg tttggggacg gccactcact cgaccactgg acaagtcgga
     4141 cacggcactg ggacacatca gatgtcggct ctgcctggtc acggaacggg acaactgacc
     4201 gaccgcgttg tggagggcac ggctgtgacc gacccgattg gaaggaacac tggaactggt
     4261 cggacaaccg ctcataacac acacgttggt ggtggtggtg ccaccgggta cggaaccggc
     4321 gggggatata ctggaagatc tatggtgtca aaaggtgagg aactgtttac aggcgtggtg
     4381 ccgatcctgg tagagctgga cggggatgtg aatggccata aattcagcgt ttcaggtgaa
     4441 ggtgagggcg acgccacgta cggaaaactg acactgaaat tcatttgcac aacaggtaaa
     4501 ctgcctgtgc cttggcctac actggtgacc acctttggtt atggtctgat gtgctttgct
     4561 cgctatcctg accacatgaa acaacatgat ttctttaaat ctgccatgcc tgaaggctac
     4621 gtgcaagagc gcaccatttt tttcaaagac gatgggaatt acaaaacacg tgcggaggtg
     4681 aaattcgagg gcgatacact ggttaaccgt atcgagctga aaggtatcga cttcaaagag
     4741 gacggaaaca ttctgggtca taaactggaa tacaactaca acagccataa cgtgtacatc
     4801 atggccgaca aacaaaaaaa cgggattaaa gtgaacttca aaatccgcca caacatcgaa
     4861 gatggcagcg tgcagctggc cgaccactat caacaaaaca caccgatcgg cgacggtcct
     4921 gtactgctgc ctgacaacca ctatctgtca tatcagagcg cactgtcaaa agatcctaac
     4981 gagaaacgcg accacatggt tctgctggaa ttcgtgacag ccgctggcat tacactgggc
     5041 atggacgagc tgtacaaata aatccagcac agtggcggcc gctcgacaat caacctctgg
     5101 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5161 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     5221 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5281 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5341 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5401 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5461 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     5521 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5581 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5641 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattctaccg ggtaggggag
     5701 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
     5761 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
     5821 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
     5881 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
     5941 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
     6001 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
     6061 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     6121 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     6181 ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg
     6241 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     6301 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     6361 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     6421 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     6481 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     6541 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     6601 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     6661 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     6721 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     6781 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
     6841 aaaggagcgc acgaccccat gcatctcgag ggcccggtac ctttaagacc aatgacttac
     6901 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctagct
     6961 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     7021 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     7081 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     7141 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     7201 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     7261 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     7321 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7381 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     7441 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7501 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     7561 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     7621 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     7681 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     7741 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     7801 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     7861 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     7921 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     7981 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     8041 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     8101 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     8161 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     8221 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     8281 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     8341 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     8401 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     8461 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     8521 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     8581 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     8641 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     8701 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     8761 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     8821 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     8881 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     8941 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     9001 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     9061 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     9121 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     9181 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     9241 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     9301 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     9361 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     9421 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     9481 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     9541 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     9601 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     9661 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     9721 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     9781 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggag
//
