LOCUS       sequence-354692-       13714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hypoxic responsive reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13714)
  AUTHORS   Allen TP, Roennfeldt AE, Reckdharajkumar M, Liu M, Quinn RJ, Russell
            DL, Peet DJ, Whitelaw ML, Bersten DC
  TITLE     dFLASH; dual FLuorescent transcription factor Activity Sensor for 
            Histone integrated live-cell reporting and high-content screening
  JOURNAL   bioRxiv 2023.11.21.568191
REFERENCE   2  (bases 1 to 13714)
  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: "bioRxiv 
            2023.11.21.568191"
COMMENT     Sequence Label: pLV-REPORT(PGK/CMV)-12xHRE
FEATURES             Location/Qualifiers
     source          1..13714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    159..180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(195..200,201..218,219..225)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    233..249
                     /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     238..260
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     257..273
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     257..273
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     292..312
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        294..312
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        340..566
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             567..747
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    794..919
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1412..1645
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1830..1874
                     /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    2136..2253
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     3295..3319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3424..4125
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(3557..3575)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3825..3844
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4019..4042
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             4135..4155
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4159..4179
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4183..4203
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4252..5379
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     CDS             5380..5433
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5428..6228
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5488..5507)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6098..6117
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6273..6296)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6623..6642
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(6745..6764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6936..7315
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7316..7519
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7470..7490
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7564..7583
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7564..7582
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7612..7636
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      7663..7672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7684..7704
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7708..7728
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7732..7752
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(7762..7764,7765..7767,7768..8478,8479..8484,
                     8485..8604)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(7807..7828)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8068..8087)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             8605..8664
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             8665..9690
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             8665..9690
                     /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"
     LTR             10923..11156
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    11228..11349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11265..11284)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11319..11338
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11451..11470
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(11735..11753)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11821..11925
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11926..11994,11995..12786)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(12144..12163)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12957..13545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13446..13465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgataaccct aattcgatag catatgcttc ccgttgggta acatatgcta ttgaattagg
       61 gttagtctgg atagtatata ctactacccg ggaagcatat gctacccgtt tagggttcac
      121 cggtgatgcc ggccacgatg cgtccggcgt agaggatcta atgtgagtta gctcactcat
      181 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      241 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      301 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
      361 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
      421 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
      481 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
      541 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
      601 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
      661 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
      721 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
      781 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
      841 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
      901 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
      961 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1021 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1081 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1141 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1201 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1261 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1321 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1381 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1441 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1501 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1561 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1621 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     1681 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     1741 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1801 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     1861 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     1921 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     1981 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2041 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2101 ccattcgatt agtgaacgga tctcgacggt atcggtttta aaagaaaagg ggggattggg
     2161 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     2221 ttacaaaaac aaattacaaa aattcaaaat tttatcgatt tcgaacctag aggacgtgac
     2281 aaacagaagc cacacgtcct agggacgtgg ggagtgcgtg aggagtacgt gaggacacgt
     2341 gggtactaga ggacgtgaca aacagaagcc acacgtccta gggacgtggg gagtgcgtga
     2401 ggagtacgtg aggacacgtg ggtactagag gacgtgacaa acagaagcca cacgtcctag
     2461 ggacgtgggg agtgcgtgag gagtacgtga ggacacgtgg gtactagagg acgtgacaaa
     2521 cagaagccac acgtcctagg gacgtgggga gtgcgtgagg agtacgtgag gacacgtggg
     2581 tactagagga cgtgacaaac agaagccaca cgtcctaggg acgtggggag tgcgtgagga
     2641 gtacgtgagg acacgtgggt actagaggac gtgacaaaca gaagccacac gtcctaggga
     2701 cgtggggagt gcgtgaggag tacgtgagga cacgtgggta ctagaggacg tgacaaacag
     2761 aagccacacg tcctagggac gtggggagtg cgtgaggagt acgtgaggac acgtgggtac
     2821 tagaggacgt gacaaacaga agccacacgt cctagggacg tggggagtgc gtgaggagta
     2881 cgtgaggaca cgtgggtact agaggacgtg acaaacagaa gccacacgtc ctagggacgt
     2941 ggggagtgcg tgaggagtac gtgaggacac gtgggtacta gaggacgtga caaacagaag
     3001 ccacacgtcc tagggacgtg gggagtgcgt gaggagtacg tgaggacacg tgggtactag
     3061 aggacgtgac aaacagaagc cacacgtcct agggacgtgg ggagtgcgtg aggagtacgt
     3121 gaggacacgt gggtactaga ggacgtgaca aacagaagcc acacgtccta gggacgtggg
     3181 gagtgcgtga ggagtacgtg aggacacgtg ggtactagag gagcatgccc atatcactag
     3241 ggtttaaacg gcgcgccagc tttaggcgtg tacggtgggc gcctataaaa gcagagctcg
     3301 tttagtgaac cgtcagatcg cctggagcaa ttccacaaca cttttgtctt ataccaactt
     3361 tccgtaccac ttcctaccct cgtaaagtcg acaccggggc ccagatctat cgatcggccg
     3421 gatatggtga gcaagggcga ggaggtcatc aaagagttca tgcgcttcaa ggtgcgcatg
     3481 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     3541 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
     3601 gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca ccccgccgac
     3661 atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     3721 ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga cggcacgctg
     3781 atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     3841 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     3901 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     3961 accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt ggacaccaag
     4021 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag
     4081 ggccgccacc acctgttcct gtacggcatg gacgagctgt acaaggccgt ggatccaaaa
     4141 aagaagagaa aggtagatcc aaaaaagaag agaaaggtag atccaaaaaa gaagagaaag
     4201 gtacacggcg ccacgaagca agcaggagat gttgaagaaa accccgggcc tatggcttcg
     4261 taccccggcc atcaacacgc gtctgcgttc gaccaggctg cgcgttctcg cggccatagc
     4321 aaccgacgta cggcgttgcg ccctcgccgg cagcaagaag ccacggaagt ccgcccggag
     4381 cagaaaatgc ccacgctact gcgggtttat atagacggtc cccacgggat ggggaaaacc
     4441 accaccacgc aactgctggt ggccctgggt tcgcgcgacg atatcgtcta cgtacccgag
     4501 ccgatgactt actggcgggt gctgggggct tccgagacaa tcgcgaacat ctacaccaca
     4561 caacaccgcc tcgaccaggg tgagatatcg gccggggacg cggcggtggt aatgacaagc
     4621 gcccagataa caatgggcat gccttatgcc gtgaccgacg ccgttctggc tcctcatatc
     4681 gggggggagg ctgggagctc acatgccccg cccccggccc tcaccctcat cttcgaccgc
     4741 catcccatcg ccgccctcct gtgctacccg gccgcgcggt accttatggg cagcatgacc
     4801 ccccaggccg tgctggcgtt cgtggccctc atcccgccga ccttgcccgg caccaacatc
     4861 gtgcttgggg cccttccgga ggacagacac atcgaccgcc tggccaaacg ccagcgcccc
     4921 ggcgagcggc tggacctggc tatgctggct gcgattcgcc gcgtttacgg gctacttgcc
     4981 aatacggtgc ggtatctgca gtgcggcggg tcgtggcggg aggactgggg acagctttcg
     5041 gggacggccg tgccgcccca gggtgccgag ccccagagca acgcgggccc acgaccccat
     5101 atcggggaca cgttatttac cctgtttcgg gcccccgagt tgctggcccc caacggcgac
     5161 ctgtataacg tgtttgcctg ggccttggac gtcttggcca aacgcctccg ttccatgcac
     5221 gtctttatcc tggattacga ccaatcgccc gccggctgcc gggacgccct gctgcaactt
     5281 acctccggga tggtccagac ccacgtcacc acccccggct ccataccgac gatatgcgac
     5341 ctggcgcgca cgtttgcccg ggagatgggg gaggctaacg agggcagagg aagtcttcta
     5401 acatgcggtg acgtggagga gaatcccggc cctatgattg aacaagatgg attgcacgca
     5461 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5521 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5581 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5641 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     5701 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5761 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5821 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5881 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5941 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgatg atctcgtcgt gacccatggc
     6001 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6061 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6121 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6181 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaaa ttctaccggg
     6241 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     6301 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     6361 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     6421 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     6481 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     6541 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcaggggcgg ggcgggcgcc
     6601 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     6661 ttctcctctt cctcatctcc gggcctttcg acctgcatct cgacggatcg ggagatctcc
     6721 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     6781 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     6841 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     6901 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     6961 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7021 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7081 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7141 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7201 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7261 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     7321 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     7381 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     7441 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     7501 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     7561 aattaatacg actcactata gggagaccca agctggctag ttaagctatc aacaagtttg
     7621 tacaaaaaag caggctttaa aggaaccaat tcagtcgagg tcgccaccat ggtgcacgtg
     7681 gatccaaaaa agaagagaaa ggtagatcca aaaaagaaga gaaaggtaga tccaaaaaag
     7741 aagagaaagg tacacgtgtc catggtgagc aagggcgagg agctgttcac cggggtggtg
     7801 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     7861 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     7921 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     7981 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     8041 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     8101 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     8161 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     8221 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     8281 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     8341 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     8401 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     8461 atggacgagc tgtacaagaa gcttagccat ggcttcccgc cggaggtgga ggagcaggat
     8521 gatggcacgc tgcccatgtc ttgtgcccag gagagcggga tggaccgtca ccctgcagcc
     8581 tgtgcttctg ctaggatcaa tgtgcagtgt actaattatg ctctcttgaa attggctgga
     8641 gatgttgaga gcaacccagg tcccatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     8701 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     8761 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     8821 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     8881 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     8941 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     9001 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     9061 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     9121 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     9181 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     9241 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     9301 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     9361 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     9421 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     9481 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     9541 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     9601 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     9661 cccagcactc gtccgagggc aaaggaatag gctagcctag tgccatttgt tcagtggttc
     9721 gtagggcttt cccccactgt ttggctttca gttatatgga tgatgtggta ttgggggcca
     9781 agtctgtaca gcatcttgag tcccttttta ccgctgttac caattttctt ttgtctttgg
     9841 gtatacattt aaaccctaac aaaacaaaga gatggggtta ctctctaaat tttatgggtt
     9901 atgtcattgg atgttatggg tccttgccac aagaacacat catacaaaaa atcaaagaat
     9961 gttttagaaa acttcctatt aacaggccta ttgattggaa agtatgtcaa cgaattgtgg
    10021 gtcttttggg ttttgctgcc ccttttacac aatgtggtta tcctgcgttg atgcctttgt
    10081 atgcatgtat tcaatctaag caggctttca ctttctcgcc aacttacaag gcctttctgt
    10141 gtaaacaata cctgaacctt taccccgttg cccggcaacg gccaggtctg tgccaagtgt
    10201 ttgctgacgc aacccccact ggctggggct tggtcatggg ccatcagcgc atgcgtggaa
    10261 ccttttcggc tcctctgccg atccatactg cggaactcct agccgcttgt tttgctcgca
    10321 gcaggtctgg agcaaacatt atcgggactg ataactctgt tgtcctatcc cgcaaatata
    10381 catcgtttcc atggctgcta ggctgtgctg ccaactggat cctgcgcggg acgtcctttg
    10441 tttacgtccc gtcggcgctg aatcctgcgg acgacccttc tcggggtcgc ttgggactct
    10501 ctcgtcccct tctccgtctg ccgttccgac cgaccacggg gcgcacctct ctttacgcgg
    10561 actccccgtc tgtgccttct catctgccgg accgtgtgca cttcgcttca cctctgcacg
    10621 tcgcatggag accaccgtga acgcccacca aatattgccc aaggtcttac ataagaggac
    10681 tcttggactc tcagcaatgt caacgaccga ccttgaggca tacttcaaag actgtttgtt
    10741 taaagactgg gaggagttgg gggaggagat taggttaaag gtctttgtac taggaggctg
    10801 taggcataaa ttggtctgcg caccagcacc atggcgcaat cactagagcg gggtaccttt
    10861 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
    10921 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
    10981 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    11041 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    11101 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
    11161 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
    11221 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    11281 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    11341 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta
    11401 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
    11461 gaggccgagg ccggatccct tgagtggctt tcatcctgga gcagactttg cagtctgtgg
    11521 actgcaacac aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg
    11581 ggggacatgt acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca
    11641 gaggggcctg tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta
    11701 taaggccccc ttgttaattc ttgaagacga aagggcctcg tgatacgcct atttttatag
    11761 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
    11821 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
    11881 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
    11941 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
    12001 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
    12061 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
    12121 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
    12181 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
    12241 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
    12301 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
    12361 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
    12421 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
    12481 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
    12541 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    12601 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    12661 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    12721 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    12781 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    12841 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    12901 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    12961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
    13021 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
    13081 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
    13141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
    13201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
    13261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    13321 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    13381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    13441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    13501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    13561 gcctttttac ggttcctggc cttttgctgg ccttgaagct gtccctgatg gtcgtcatct
    13621 acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga
    13681 atcataatgg ggaaggccat ccagcctcgc gtcg
//
