LOCUS       sequence-224742-       11654 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NanoLuc-Firefly Luciferase fusion with constitutive nuclear 
            expression for calibration of Nanoluc signal to Firefly Luciferase 
            signal. H2B-mCherry marks chromatin with red fluorescence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11654)
  AUTHORS   Mandic A, Strebinger D, Regali C, Phillips NE, Suter DM
  TITLE     A novel method for quantitative measurements of gene expression in 
            single living cells.
  JOURNAL   Methods. 2017 May 1;120:65-75. doi: 10.1016/j.ymeth.2017.04.008. 
            Epub 2017 Apr 27.
  PUBMED    28456689
REFERENCE   2  (bases 1 to 11654)
  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: "Methods."; 
            date: "2017-05-1"; volume: "120:65-75. doi"; pages: " 
            10.1016/j.ymeth.2017.04.008. Epub 2017 Apr 27"
COMMENT     Sequence Label: pLV-pGK-Nluc-Fluc-NLS-P2A-H2B-mCherry-IRES-Hygro
FEATURES             Location/Qualifiers
     source          1..11654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..234
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    306..427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(343..362)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     397..416
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      446..581
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(613..632)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(614..632)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(642..659)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(642..658)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(651..673)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      803..1258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(890..909)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1100..1121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1287..1391
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1392..1460,1461..2252)
                     /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(1610..1629)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2423..3011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2912..2931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3165..3182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3299..3320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3335..3340,3341..3358,3359..3365)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3373..3389
                     /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     3378..3400
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3397..3413
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3397..3413
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3432..3452
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3434..3452
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             3708..3888
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3932..4057
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4550..4783
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4968..5012
                     /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    5279..5396
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5443..5953
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5872..5890
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             5962..6474
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             6481..8130
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     CDS             8140..8160
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8176..8232
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8239..8616
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKDGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAVTKYTSAK"
     CDS             8623..9333
                     /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(8768..8786)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9036..9055
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    join(9353..9927,9928..9930,9931..9939)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9520..9537)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9747..9766
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             9940..10965
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             9940..10965
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    10980..11568
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11033..11053)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11451..11462)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
       61 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      121 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      181 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
      241 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
      301 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
      361 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
      421 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
      481 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
      541 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcgtcgagac
      601 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
      661 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
      721 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
      781 agcctgaatg gcgaatggcg cgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      841 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      901 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      961 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1021 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1081 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1141 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1201 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttcc
     1261 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     1321 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1381 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1441 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1501 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1561 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1621 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1681 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1741 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1801 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1861 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1921 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1981 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2041 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2101 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2161 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2221 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2281 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     2341 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2401 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2461 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2521 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2581 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2641 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2701 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2761 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2821 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2881 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2941 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3001 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3061 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3121 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3181 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3241 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3301 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     3361 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     3421 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     3481 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     3541 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     3601 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     3661 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt
     3721 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     3781 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     3841 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     3901 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga
     3961 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     4021 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     4081 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     4141 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     4201 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     4261 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     4321 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     4381 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     4441 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     4501 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     4561 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg
     4621 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     4681 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     4741 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     4801 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     4861 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     4921 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     4981 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     5041 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     5101 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     5161 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
     5221 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt
     5281 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     5341 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttatcg
     5401 atcacgagac tagcctcgag aagcttgata tcgaattccc acggggttgg ggttgcgcct
     5461 tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa
     5521 acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg
     5581 gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg
     5641 ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac
     5701 tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc
     5761 tgtggccaat agcggctgct cagcggggcg cgccgagagc agcggccggg aaggggcggt
     5821 gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc
     5881 attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga
     5941 cctctctccc cagggggatc catggtcttc acactcgaag atttcgttgg ggactggcga
     6001 cagacagccg gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc cagtttgttt
     6061 cagaatctcg gggtgtccgt aactccgatc caaaggattg tcctgagcgg tgaaaatggg
     6121 ctgaagatcg acatccatgt catcatcccg tatgaaggtc tgagcggcga ccaaatgggc
     6181 cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg atcatcactt taaggtgatc
     6241 ctgcactatg gcacactggt aatcgacggg gttacgccga acatgatcga ctatttcgga
     6301 cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac agggaccctg
     6361 tggaacggca acaaaattat cgacgagcgc ctgatcaacc ccgacggctc cctgctgttc
     6421 cgagtaacca tcaacggagt gaccggctgg cggctgtgcg aacgcattct ggcggtcgac
     6481 atggaagatg ccaaaaacat taagaagggc ccagcgccat tctacccact cgaagacggg
     6541 accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc cggcaccatc
     6601 gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt cgagatgagc
     6661 gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg gatcgtggtg
     6721 tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg
     6781 gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag catgggcatc
     6841 agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa
     6901 aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga ctaccagggc
     6961 ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa cgagtacgac
     7021 ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa cagtagtggc
     7081 agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt
     7141 catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat cctcagcgtg
     7201 gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat ctgcggcttt
     7261 cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt gcaagactat
     7321 aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa gagcactctc
     7381 atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc gccgctcagc
     7441 aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg ccagggctac
     7501 ggcctgacag aaacaaccag cgccattctg atcacccccg aaggggacga caagcctggc
     7561 gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga caccggtaag
     7621 acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat catgagcggc
     7681 tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg gctgcacagc
     7741 ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg gctgaagagc
     7801 ctgatcaaat acaagggcta ccaggtagcc ccagccgaac tggagagcat cctgctgcaa
     7861 caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg
     7921 cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga gatcgtggac
     7981 tatgtggcca gccaggttac aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac
     8041 gaggtgccta aaggactgac cggcaagttg gacgcccgca agatccgcga gattctcatt
     8101 aaggccaaga agggcggcaa gatcgccgtg aattggcctc caaaaaagaa gagaaaggta
     8161 atcgatggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
     8221 aaccctggac cttctagaat gccagagcca gcgaagtctg ctcccgcccc gaaaaagggc
     8281 tccaagaagg cggtgactaa ggcgcagaag aaagacggca agaagcgcaa gcgcagccgc
     8341 aaggagagct attccatcta tgtgtacaag gttctgaagc aggtccaccc tgacaccggc
     8401 atttcgtcca aggccatggg catcatgaat tcgtttgtga acgacatttt cgagcgcatc
     8461 gcaggtgagg cttcccgcct ggcgcattac aacaagcgct cgaccatcac ctccagggag
     8521 atccagacgg ccgtgcgcct gctgctgcct ggggagttgg ccaagcacgc cgtgtccgag
     8581 ggtactaagg ccgtcaccaa gtacaccagc gctaaggatc cgatggtgag caagggcgag
     8641 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     8701 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     8761 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     8821 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     8881 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     8941 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     9001 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     9061 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     9121 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     9181 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     9241 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     9301 tccaccggcg gcatggacga gctgtacaag taacccggga gaataaggac cgcccctctc
     9361 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     9421 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     9481 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     9541 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     9601 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     9661 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     9721 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     9781 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     9841 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     9901 ttttcctttg aaaaacacga tgataatatg gccacaacca tgaaaaagcc tgaactcacc
     9961 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
    10021 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
    10081 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
    10141 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg
    10201 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
    10261 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
    10321 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
    10381 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
    10441 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
    10501 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
    10561 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
    10621 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
    10681 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
    10741 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
    10801 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
    10861 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
    10921 agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagtaagc tagctcgaca
    10981 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
    11041 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
    11101 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
    11161 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
    11221 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
    11281 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
    11341 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
    11401 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
    11461 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
    11521 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag
    11581 ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
    11641 agaaaagggg ggac
//
