LOCUS       sequence-495607-       12314 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Overexpression vector for eGFP tagged ESR1 or ER-alpha. Monitor 
            post-translational degradation of ER-alpha via EGFP:mCherry ratio. 
            Contains puromycin-resistance selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12314)
  AUTHORS   Tsai JM, Aguirre JD, Li YD, Brown J, Focht V, Kater L, Kempf G, 
            Sandoval B, Schmitt S, Rutter JC, Galli P, Sandate CR, Cutler JA, 
            Zou C, Donovan KA, Lumpkin RJ, Cavadini S, Park PMC, Sievers Q, 
            Hatton C, Ener E, Regalado BD, Sperling MT, Slabicki M, Kim J, Zon 
            R, Zhang Z, Miller PG, Belizaire R, Sperling AS, Fischer ES, 
            Irizarry R, Armstrong SA, Thoma NH, Ebert BL
  TITLE     UBR5 forms ligand-dependent complexes on chromatin to regulate 
            nuclear hormone receptor stability.
  JOURNAL   Mol Cell. 2023 Aug 3;83(15):2753-2767.e10. doi: 
            10.1016/j.molcel.2023.06.028. Epub 2023 Jul 20.
  PUBMED    37478846
REFERENCE   2  (bases 1 to 12314)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2023.06.028"; journalName: "Mol Cell"; date: 
            "2023-08-3- 3"; volume: "83"; issue: "15"; pages: "2753-2767.e10"
COMMENT     Sequence Label: pZucchini2_SFFV-ESR1-eGFP-IRES-mCherry_PuroR
FEATURES             Location/Qualifiers
     source          1..12314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             42..86
                     /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"
     promoter        288..694
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             join(2553..2555,2556..3269)
                     /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(2553..2555,2556..3266)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2595..2616)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2856..2875)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3203..3224
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(3293..3867,3868..3870,3871..3879)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3460..3477)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3687..3706
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3889..3898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3895..4605
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4040..4058)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4308..4327
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4661..4778
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4912..6090
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5143..6081
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6038..6058
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             6103..6702
                     /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(6103..6122)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6599..6619
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6718..7306
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6771..6791)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7189..7200)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7378..7611
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7689..7810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7726..7745)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7780..7799
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7850..7985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8005..8024)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8006..8024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8034..8051)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8034..8050)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8043..8065)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8192..8647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8279..8298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8489..8510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8673..8777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8778..8846,8847..9638)
                     /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(8996..9015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9809..10397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10298..10317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10551..10568
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10685..10706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10721..10726,10727..10744,10745..10751)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10759..10775
                     /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     10764..10786
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10783..10799
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10783..10799
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10818..10838
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10820..10838
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10866..11092
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11093..11273
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11320..11445
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11938..12171
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
       61 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
      121 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
      181 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
      241 tcccaacccc gaggggaccc tattccagca catatgagct agctgcagta acgccatttt
      301 gcaaggcatg gaaaaatacc aaaccaagaa tagagaagtt cagatcaagg gcgggtacat
      361 gaaaatagct aacgttgggc caaacaggat atctgcggtg agcagtttcg gccccggccc
      421 ggggccaaga acagatggtc accgcagttt cggccccggc ccgaggccaa gaacagatgg
      481 tccccagata tggcccaacc ctcagcagtt tcttaagacc catcagatgt ttccaggctc
      541 ccccaaggac ctgaaatgac cctgcgcctt atttgaatta accaatcagc ctgcttctcg
      601 cttctgttcg cgcgcttctg cttcccgagc tctataaaag agctcacaac ccctcactcg
      661 gcgcgccagt cctccgacag actgagtcgg ccggtcgaat caagcttatc gataccgtcg
      721 actccggaat agccaccatg acaatgactc tgcatacgaa ggcctcaggc atggcattgc
      781 tccaccaaat tcagggcaat gaactcgaac ctttgaatcg cccacaactt aaaataccat
      841 tggaacgccc tttgggcgaa gtctatttgg attcttctaa accagcggta tataattatc
      901 cagaaggggc tgcttatgaa tttaatgcgg ctgcggctgc taatgctcaa gtgtatggcc
      961 aaactggact gccttatggg cctggaagtg aagccgccgc ctttgggtct aatggactcg
     1021 gcggatttcc acctctgaat tctgtctccc ccagccctct catgttgctt catccaccac
     1081 cccaattgag ccccttcttg caaccacatg ggcaacaagt cccatattat cttgaaaatg
     1141 aacctagtgg ttataccgtt agagaagctg gtccaccagc tttctataga cccaacagtg
     1201 acaaccgcag acaaggcggt agggagcgac tggcgtcaac aaacgataaa ggctccatgg
     1261 ccatggagtc agctaaagaa acacgatatt gcgccgtttg taacgattac gcgtctggat
     1321 atcactacgg cgtttggtca tgcgaaggtt gtaaagcatt ctttaaacgg agcatccagg
     1381 gtcacaatga ttacatgtgc ccggctacaa atcaatgtac aatcgacaag aatcgacgca
     1441 aatcatgtca agcttgtaga ttgcggaagt gttatgaggt tggcatgatg aagggtggca
     1501 ttaggaagga tcgtcgcggt ggccgcatgc tgaagcataa acggcaaagg gacgacggcg
     1561 aaggacgcgg cgaggtaggt tcagccggtg atatgcgggc cgctaatctc tggccttctc
     1621 cactgatgat taagcggagc aagaagaata gtctcgcact gtcactcaca gcagatcaaa
     1681 tggtgtccgc gctgctggac gccgaaccac ccatcctgta ctcagaatac gacccaacac
     1741 gcccgtttag cgaggcgtct atgatgggac tgctcacaaa tctcgcggat cgggaactcg
     1801 tgcatatgat taattgggct aaaagagtcc ccggtttcgt cgacctcaca ctgcacgacc
     1861 aagttcatct gttggagtgc gcatggctgg aaattttgat gatcggactt gtgtggcgat
     1921 caatggaaca tccgggaaaa ctgcttttcg ccccaaatct tctgctcgat agaaatcaag
     1981 gcaagtgcgt ggaaggaatg gttgaaatat ttgatatgct cctcgcaacc agtagtagat
     2041 ttaggatgat gaacttgcaa ggggaagaat tcgtttgtct gaagagtatc atcctgctga
     2101 acagtggcgt atataccttc ctctcatcca cactcaaatc cctcgaggag aaagatcaca
     2161 ttcatagggt gctcgataaa attacggata ccctgattca tctcatggct aaagccgggc
     2221 tcacgttgca acaacaacat caacgcctcg cacaactgct gctgattctg tcacatatac
     2281 gacatatgag caataaggga atggaacacc tctattctat gaaatgtaag aatgtcgtac
     2341 ctctgtacga tcttttgctt gaaatgttgg atgcacatag gctccacgct cctacctcaa
     2401 ggggcggagc ctcagtagaa gagactgatc agtcccatct ggctacagca gggtccacca
     2461 gtagtcacag cctgcagaaa tactatatta caggcgaagc cgaagggttt cccgcgactg
     2521 tgggcggcgg cggctccggc ggcggcggct ccgtgagcaa gggcgaggag ctgttcaccg
     2581 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2641 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2701 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2761 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2821 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2881 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2941 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3001 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3061 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3121 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3181 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3241 ctctcggcat ggacgagctg tacaagtaaa tgcatgagta actgaggatc cgcccctctc
     3301 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     3361 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     3421 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     3481 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     3541 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     3601 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     3661 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     3721 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     3781 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3841 ttttcctttg aaaaacacga tgataatatg gccacaaccc tggaattcgc caccatggtg
     3901 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     3961 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     4021 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     4081 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     4141 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     4201 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     4261 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     4321 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     4381 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     4441 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     4501 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     4561 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaactag taagcttggc
     4621 gtaactagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     4681 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     4741 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     4801 gagatccact ttgggctcga gggggcccgg gtgcaaagat ggataaagtt ttaaacagag
     4861 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
     4921 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     4981 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     5041 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     5101 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     5161 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
     5221 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
     5281 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
     5341 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     5401 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     5461 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     5521 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     5581 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     5641 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     5701 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     5761 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     5821 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     5881 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     5941 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     6001 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     6061 agtttttttc ttccatttca ggtgtcgtga cgtacggcca ccatgaccga gtacaagccc
     6121 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     6181 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     6241 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     6301 gcggacgacg gcgccgccgt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     6361 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     6421 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     6481 accgtcggag tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     6541 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     6601 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     6661 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat
     6721 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     6781 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     6841 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     6901 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     6961 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     7021 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     7081 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     7141 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     7201 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     7261 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     7321 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     7381 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     7441 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7501 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7561 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtacgtata
     7621 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
     7681 gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     7741 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     7801 atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct
     7861 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     7921 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     7981 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     8041 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     8101 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     8161 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     8221 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     8281 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     8341 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     8401 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     8461 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     8521 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     8581 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     8641 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     8701 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     8761 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     8821 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     8881 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     8941 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     9001 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     9061 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     9121 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     9181 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     9241 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     9301 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     9361 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     9421 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     9481 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     9541 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     9601 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     9661 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     9721 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     9781 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     9841 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     9901 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     9961 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
    10021 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
    10081 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
    10141 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
    10201 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
    10261 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
    10321 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
    10381 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
    10441 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
    10501 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
    10561 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
    10621 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
    10681 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
    10741 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
    10801 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
    10861 cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
    10921 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
    10981 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
    11041 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
    11101 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
    11161 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
    11221 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
    11281 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
    11341 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
    11401 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
    11461 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
    11521 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
    11581 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
    11641 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
    11701 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
    11761 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
    11821 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
    11881 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
    11941 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
    12001 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
    12061 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
    12121 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
    12181 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
    12241 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
    12301 caattacaca agct
//
