LOCUS       sequence-385702-       12781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline inducible expression of short Lepr (no signaling) with 
            eGFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12781)
  AUTHORS   Yuan S, Stewart KS, Yang Y, Abdusselamoglu MD, Parigi SM, Feinberg 
            TY, Tumaneng K, Yang H, Levorse JM, Polak L, Ng D, Fuchs E
  TITLE     Ras drives malignancy through stem cell crosstalk with the 
            microenvironment.
  JOURNAL   Nature. 2022 Nov 30. doi: 10.1038/s41586-022-05475-6.
  PUBMED    36450983
REFERENCE   2  (bases 1 to 12781)
  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: "Nature. 
            2022 Nov 30. doi: 10.1038/s41586-022-05475-6."
COMMENT     Sequence Label: TRE-Lepr_Short-IRES-eGFP
FEATURES             Location/Qualifiers
     source          1..12781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             2..635
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    682..807
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1300..1533
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1718..1762
                     /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"
     protein_bind    join(2067..2085,2086..2108,2109..2127,2128..2150,2151..2169,
                     2170..2192,2193..2211,2212..2234,2235..2253,2254..2276,
                     2277..2295,2296..2318,2319..2337)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2370..2408
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2405..2429
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    2600..2717
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5215..5782
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5233..5784
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5377..5394)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5604..5623
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5787..5789,5790..5792,5793..6503)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5787..5789,5790..5792,5793..6503)
                     /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(5832..5853)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6093..6112)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6440..6461
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6582..6598)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6583..6599)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     misc_feature    6612..7200
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6665..6685)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7083..7094)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(7344..7377)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             7397..7577
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7602..7620)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7603..7620)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7910..7929)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(8143..8162)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8262..8284
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8322..8340)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8408..8512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8513..8581,8582..9373)
                     /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(8731..8750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9544..10132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10033..10052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10286..10303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(10373..10393)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10378..10707
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10558..10693
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          11841..11906
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             12036..12056
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    12481..12615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12518..12537)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12572..12591
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg gatctaccac
       61 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
      121 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
      181 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
      241 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
      301 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
      361 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
      421 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
      481 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      541 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      601 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      661 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      721 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      781 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      841 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      901 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
      961 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1021 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1081 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1141 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1201 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1261 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1321 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1381 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1441 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1501 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1561 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1621 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1681 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1741 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1801 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1861 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1921 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     1981 agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg
     2041 accatcgcta gagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     2101 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     2161 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     2221 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
     2281 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
     2341 gtgaaagtcg agctcggtac ccgggtcgag gtaggcgtgt acggtgggag gcctatataa
     2401 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2461 tccatagaag acaccgggac cgatccagcc tccgcggccc cgaattgtga gatctcaacc
     2521 ggtttaggcg gccgcttctg ctagctaatt cgagctcggt acccggggat cctctagaat
     2581 ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac agtgcagggg
     2641 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     2701 caaaaattca aaattttatc gatcacgaga ctagcctcga ggtttcgcca ccatgatgtg
     2761 tcagaaattc tatgtggttt tgttacactg ggaatttctt tatgtgatag ctgcacttaa
     2821 cctggcatat ccaatctctc cctggaaatt taagttgttt tgtggaccac cgaacacaac
     2881 cgatgactcc tttctctcac ctgctggagc cccaaacaat gcctcggctt tgaagggggc
     2941 ttctgaagca attgttgaag ctaaatttaa ttcaagtggt atctacgttc ctgagttatc
     3001 caaaacagtc ttccactgtt gctttgggaa tgagcaaggt caaaactgct ctgcactcac
     3061 agacaacact gaagggaaga cactggcttc agtagtgaag gcttcagttt ttcgccagct
     3121 aggtgtaaac tgggacatag agtgctggat gaaaggggac ttgacattat tcatctgtca
     3181 tatggagcca ttacctaaga accccttcaa gaattatgac tctaaggtcc atcttttata
     3241 tgatctgcct gaagtcatag atgattcgcc tctgccccca ctgaaagaca gctttcagac
     3301 tgtccaatgc aactgcagtc ttcggggatg tgaatgtcat gtgccggtac ccagagccaa
     3361 actcaactac gctcttctga tgtatttgga aatcacatct gccggtgtga gttttcagtc
     3421 acctctgatg tcactgcagc ccatgcttgt tgtgaaaccc gatccaccct taggtttgca
     3481 tatggaagtc acagatgatg gtaatttaaa gatttcttgg gacagccaaa caatggcacc
     3541 atctccgctt caatatcagg tgaaatattt agagaattct acaattgtaa gagaggctgc
     3601 tgaaattgtc tcagctacat ctctgctggt agacagtgtg cttcctggat cttcatatga
     3661 ggtccaggtg aggagcaaga gactggatgg ttcaggagtc tggagtgact ggagttcacc
     3721 tcaagtcttt accacacaag atgttgtgta ttttccaccc aaaattctga ctagtgttgg
     3781 atcgaatgct tcttttcatt gcatctacaa aaacgaaaac cagattatct cctcaaaaca
     3841 gatagtttgg tggaggaatc tagctgagaa aatccctgag atacagtaca gcattgtgag
     3901 tgaccgagtt agcaaagtta ccttctccaa cctgaaagcc accagacctc gagggaagtt
     3961 tacctatgac gcagtgtact gctgcaatga gcaggcgtgc catcaccgct atgctgaatt
     4021 atacgtgatc gatgtcaata tcaatatatc atgtgaaact gacgggtact taactaaaat
     4081 gacttgcaga tggtcaccca gcacaatcca atcactagtg ggaagcactg tgcagctgag
     4141 gtatcacagg cgcagcctgt attgtcctga tagtccatct attcatccta cgtctgagcc
     4201 caaaaactgc gtcttacaga gagacggctt ttatgaatgt gttttccagc caatctttct
     4261 attatctggc tatacaatgt ggatcaggat caaccattct ttaggttcac ttgactcgcc
     4321 accaacgtgt gtccttcctg actccgtagt aaaaccacta cctccatcta acgtaaaagc
     4381 agagattact gtaaacactg gattattgaa agtatcttgg gaaaagccag tctttccgga
     4441 gaataacctt caattccaga ttcgatatgg cttaagtgga aaagaaatac aatggaagac
     4501 acatgaggta ttcgatgcaa agtcaaagtc tgccagcctg ctggtgtcag acctctgtgc
     4561 agtctatgtg gtccaggttc gctgccggcg gttggatgga ctaggatatt ggagtaattg
     4621 gagcagtcca gcctatacgc ttgtcatgga tgtaaaagtt cctatgagag ggcctgaatt
     4681 ttggagaaaa atggatgggg acgttacctt ctcatggccc atgagtaaag tgagtgctgt
     4741 ggagtcactc agtgcttatc ccctgagcag cagctgtgtc atcctttcct ggacactgtc
     4801 acctgatgat tatagtctgt tatatctggt tattgaatgg aagatcctta atgaagatga
     4861 tggaatgaag tggcttagaa ttccctcgaa tgttaaaaag ttttatatcc acgataattt
     4921 tattcccatc gagaaatatc agtttagtct ttacccagta tttatggaag gagttggaaa
     4981 accaaagata attaatggtt tcaccaaaga tgctatcgac aagcagcaga atgacgcagg
     5041 gctgtatgtc attgtaccca taattatttc ctcttgtgtc ctactgctcg gaacactgtt
     5101 aatttcacac cagagaatga aaaagttgtt ttgggacgat gttccaaacc ccaagaattg
     5161 ttcctgggca caaggactga atttccaaaa gagaacggac actctttgag gaattccgcc
     5221 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     5281 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     5341 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     5401 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     5461 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     5521 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     5581 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     5641 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     5701 catgtgttta gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     5761 cctttgaaaa acacgatgat aataccatgg tgagcaaggg cgaggagctg ttcaccgggg
     5821 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     5881 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     5941 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     6001 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     6061 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     6121 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     6181 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     6241 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     6301 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     6361 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     6421 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     6481 tcggcatgga cgagctgtac aagtccggac tcagatctcg actagctagt agctagctag
     6541 ctagtcgagc tcaagcttcg aattcgatat caagcttatc gcgataccgt cgacctcgag
     6601 ggaattccga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6661 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6721 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6781 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6841 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6901 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6961 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     7021 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     7081 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     7141 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     7201 atcgggaatt cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     7261 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     7321 gatgggatca attcaccatg ggaataactt cgtatagcat acattatacg aagttatgct
     7381 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     7441 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     7501 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     7561 gtggaaaatc tctagcagca tctagaatta attccgtgta ttctatagtg tcacctaaat
     7621 cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg
     7681 tacaagccta attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa
     7741 actgccgtca gagtcggttt ggttggacga accttctgag tttctggtaa cgccgtcccg
     7801 cacccggaaa tggtcagcga accaatcagc agggtcatcg ctagccagat cctctacgcc
     7861 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     7921 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     7981 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     8041 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     8101 caggagtcgc ataagggaga gcgtcgaatg gtgcactctc agtacaatct gctctgatgc
     8161 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     8221 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     8281 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     8341 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     8401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     8461 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     8521 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     8581 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     8641 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     8701 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     8761 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8821 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8881 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8941 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     9001 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     9061 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     9121 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     9181 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     9241 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     9301 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     9361 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     9421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     9481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     9541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     9601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     9661 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     9721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     9841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     9901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     9961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
    10021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
    10081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
    10141 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
    10201 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
    10261 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
    10321 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct gtggaatgtg
    10381 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10441 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
    10501 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
    10561 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
    10621 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
    10681 ttttttggag gcctaggctt ttgcaaaaag cttggacaca agacaggctt gcgagatatg
    10741 tttgagaata ccactttatc ccgcgtcagg gagaggcagt gcgtaaaaag acgcggactc
    10801 atgtgaaata ctggttttta gtgcgccaga tctctataat ctcgcgcaac ctattttccc
    10861 ctcgaacact ttttaagccg tagataaaca ggctgggaca cttcacatga gcgaaaaata
    10921 catcgtcacc tgggacatgt tgcagatcca tgcacgtaaa ctcgcaagcc gactgatgcc
    10981 ttctgaacaa tggaaaggca ttattgccgt aagccgtggc ggtctgtacc gggtgcgtta
    11041 ctggcgcgtg aactgggtat tcgtcatgtc gataccgttt gtatttccag ctacgatcac
    11101 gacaaccagc gcgagcttaa agtgctgaaa cgcgcagaag gcgatggcga aggcttcatc
    11161 gttattgatg acctggtgga taccggtggt actgcggttg cgattcgtga aatgtatcca
    11221 aaagcgcact ttgtcaccat cttcgcaaaa ccggctggtc gtccgctggt tgatgactat
    11281 gttgttgata tcccgcaaga tacctggatt gaacagccgt gggatatggg cgtcgtattc
    11341 gtcccgccaa tctccggtcg ctaatctttt caacgcctgg cactgccggg cgttgttctt
    11401 tttaacttca ggcgggttac aatagtttcc agtaagtatt ctggaggctg catccatgac
    11461 acaggcaaac ctgagcgaaa ccctgttcaa accccgcttt aaacatcctg aaacctcgac
    11521 gctagtccgc cgctttaatc acggcgcaca accgcctgtg cagtcggccc ttgatggtaa
    11581 aaccatccct cactggtatc gcatgattaa ccgtctgatg tggatctggc gcggcattga
    11641 cccacgcgaa atcctcgacg tccaggcacg tattgtgatg agcgatgccg aacgtaccga
    11701 cgatgattta tacgatacgg tgattggcta ccgtggcggc aactggattt atgagtgggc
    11761 cccggatctt tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta
    11821 cagagattta aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac
    11881 tgattctaat tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt
    11941 ggtggaatgc ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg
    12001 atgaggctac tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag
    12061 accccaagga ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata
    12121 gaactcttgc ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga
    12181 aaattatgga aaaatattct gtaaccttta taagtaggca taacagttat aatcataaca
    12241 tactgttttt tcttactcca cacaggcata gagtgtctgc tattaataac tatgctcaaa
    12301 aattgtgtac ctttagcttt ttaatttgta aaggggttaa taaggaatat ttgatgtata
    12361 gtgccttgac tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta
    12421 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
    12481 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    12541 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    12601 tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc
    12661 ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
    12721 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
    12781 g
//
