LOCUS       sequence-178091-       13492 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of EGFP with a C-terminal NLS sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13492)
  AUTHORS   Shah M, Smolko CM, Kinicki S, Chapman ZD, Brautigan DL, Janes KA
  TITLE     Profiling Subcellular Protein Phosphatase Responses to 
            Coxsackievirus B3 Infection of Cardiomyocytes.
  JOURNAL   Mol Cell Proteomics. 2017 Apr;16(4 suppl 1):S244-S262. doi: 
            10.1074/mcp.O116.063487. Epub 2017 Feb 7.
  PUBMED    28174228
REFERENCE   2  (bases 1 to 13492)
  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: "Mol Cell 
            Proteomics."; date: "2017-04"; volume: "16(4 suppl 1):S244-S262. 
            doi"; pages: " 10.1074/mcp.O116.063487. Epub 2017 Feb 7"
COMMENT     Sequence Label: pSLIK 3XFLAG-EGFP-NLS hygro
FEATURES             Location/Qualifiers
     source          1..13492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    11..35
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        67..381
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    75..93
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    111..129
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    146..164
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    182..200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    218..236
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    253..271
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    289..307
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     354..378
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             390..455
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(462..464,465..1175)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(504..525)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(765..784)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1112..1133
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1209..1229
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(1268..1292)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        1342..2553
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2390..2409
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(2678..3295,3296..3301,3302..3337,3338..3340,3341..3376,
                     3377..3379,3380..3415,3416..3424)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    3462..4033
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3627..3644)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3854..3873
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4092..5111
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             4092..5111
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    5577..6165
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5630..5650)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6048..6059)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6168..6184)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6169..6185)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6694..6874
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6900..6917)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6906..7130
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7176..7604
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7263..7282)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7473..7494
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7613..7633)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7618..7947
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7798..7933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7860..7879
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7995..8042
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8061..8435
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8565..8686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8602..8621)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8656..8675
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8735..8751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8735..8751)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8748..8770)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8759..8775
                     /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(8783..8789,8790..8807,8808..8813))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8828..8849
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8966..8983)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9137..9725)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9217..9236)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9896..10687,10688..10756))
                     /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     10519..10538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10757..10861)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10936..10955)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11127..11506
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11508..11706
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             11724..11904
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11951..12076
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12569..12802
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12987..13031
                     /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    13329..13446
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 acagttatca acaagtttgt acaaaaaagc aggctttaaa ggaaccaatt cagtcgactg
       61 gatcccgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
      121 tgatagagaa cgatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta
      181 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt
      241 atgtcgagtt tatccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      301 atagagaacg tatgtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt
      361 ttagtgaacc gtcagatcgc cactagatgg actacaaaga ccatgacggt gattataaag
      421 atcatgacat cgattacaag gatgacgatg acaagactag tgtgagcaag ggcgaggagc
      481 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      541 tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      601 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      661 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      721 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      781 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      841 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      901 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg aacttcaaga
      961 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1021 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1081 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1141 ccgggatcac tctcggcatg gacgagctgt acaagtccaa gggtgggcgc gccgacggtg
     1201 gttctggtcc gaagaaaaaa cgtaaagtgt agcaattgga agactaatgc gttttcgaga
     1261 tatctagacc cagctttctt gtacaaagtg gttgataaca tctcgagcca taaagatggt
     1321 taattaaccc acccaagatc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     1381 ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt cctgatcctt
     1441 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     1501 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     1561 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     1621 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     1681 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     1741 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     1801 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     1861 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     1921 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     1981 aacccaaggt cttgaggcct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     2041 ggggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     2101 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     2161 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     2221 ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     2281 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     2341 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     2401 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     2461 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agactagtaa attgtccgct
     2521 aaattctggc cgtttttggc ttttttgtta gacgaagctt ggtaccgagc tcggatctcc
     2581 accccgtacc ggtcctgcag gatttaaatg gcgcgccgct agcgctaccg gactcagatc
     2641 tcgagctcaa gcttcgaatt ctgcagtcga attcaccatg tctagactgg acaagagcaa
     2701 agtcataaac ggagctctgg aattactcaa tggtgtcggt atcgaaggcc tgacgacaag
     2761 gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa
     2821 gcgggccctg ctcgatgccc tgccaatcga gatgctggac aggcatcata cccacttctg
     2881 ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cataccgctg
     2941 tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa
     3001 acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga
     3061 gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga
     3121 acaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc
     3181 acttctgaga caagcaattg agctgttcga ccggcaggga gccgaacctg ccttcctttt
     3241 cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc
     3301 gaccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt
     3361 tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg
     3421 gtaactaagt aaggatccgc ggccgcacta gaggaattcc gcccctctcc cccccccccc
     3481 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     3541 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     3601 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     3661 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     3721 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     3781 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     3841 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     3901 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     3961 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     4021 aacacgatga taagcttacc ggtacgcgtg atagcttgcc acaacccgta ccaaagatgg
     4081 atagatccgg aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4141 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4201 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4261 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     4321 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4381 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     4441 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     4501 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     4561 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     4621 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     4681 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     4741 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     4801 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     4861 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     4921 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     4981 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     5041 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5101 caaaggaata gagtagatgc cgaccgaaca agagctgatt tcgagaacgc ctcagccagc
     5161 aactcgcgcg agcctagcaa ggcaaatgcg agagaacggc cttacgcttg gtggcacagt
     5221 tctcgtccac agttcgctaa gctcgctcgg ctgggtcgcg ggagggccgg tcgcagtgat
     5281 tcaggccctt ctggattgtg ttggtcccca gggcacgatt gtcatgccca cgcactcggg
     5341 tgatctgact gatcccgcag attggagatc gccgcccgtg cctgccgatt gggtgcagat
     5401 ctctagatct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     5461 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     5521 aatgaatgca attgttgttg tttagtccct cccaattcga tatcaagctt atcgataatc
     5581 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5641 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5701 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5761 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5821 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5881 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5941 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     6001 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     6061 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     6121 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     6181 ctcgatcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     6241 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     6301 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6361 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     6421 tggatctacc acacacaagg ctacttccct gattggcaga actacacacc agggccaggg
     6481 atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag
     6541 gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg
     6601 atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat
     6661 cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga
     6721 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6781 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6841 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc
     6901 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     6961 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     7021 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     7081 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     7141 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     7201 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     7261 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     7321 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     7381 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     7441 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     7501 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     7561 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     7621 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     7681 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     7741 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     7801 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     7861 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     7921 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     7981 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     8041 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
     8101 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
     8161 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
     8221 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
     8281 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
     8341 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
     8401 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
     8461 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     8521 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     8581 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     8641 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     8701 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     8761 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     8821 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     8881 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     8941 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     9001 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     9061 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     9121 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     9181 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     9241 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     9301 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     9361 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     9421 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     9481 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     9541 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     9601 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     9661 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     9721 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     9781 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     9841 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     9901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     9961 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
    10021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    10081 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    10141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    10201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    10261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    10321 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    10381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    10441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    10501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    10561 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    10621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    10681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    10741 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    10801 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    10861 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
    10921 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
    10981 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
    11041 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
    11101 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
    11161 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
    11221 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
    11281 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
    11341 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
    11401 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
    11461 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
    11521 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
    11581 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
    11641 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
    11701 aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
    11761 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
    11821 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
    11881 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
    11941 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
    12001 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
    12061 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
    12121 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
    12181 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
    12241 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
    12301 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
    12361 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
    12421 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
    12481 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
    12541 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
    12601 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
    12661 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
    12721 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
    12781 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
    12841 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
    12901 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
    12961 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
    13021 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
    13081 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
    13141 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
    13201 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
    13261 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
    13321 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
    13381 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
    13441 aattttcggg tttattacag ggacagcaga gatccagttt ggttaattct ag
//
