LOCUS       Exported               10646 bp ds-DNA     circular SYN 16-APR-2024
DEFINITION  Lentiviral expression of the cell cycle marker PIP-FUCCI.
ACCESSION   .
VERSION     .
KEYWORDS    pLenti-PGK-Hygro-PIP-FUCCI
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10646)
  AUTHORS   Grant GD, Kedziora KM, Limas JC, Cook JG, Purvis JE
  TITLE     Accurate delineation of cell cycle phase transitions in living cells
            with PIP-FUCCI.
  JOURNAL   Cell Cycle. 2018;17(21-22):2496-2516. doi: 
            10.1080/15384101.2018.1547001.
  PUBMED    30421640
REFERENCE   2  (bases 1 to 10646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 16, 2024 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10646
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    45..633
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(98..118)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(516..527)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        689..1188
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(724..747)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1121..1140
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1206..2231
                     /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             2605..2838
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2910..3031
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2947..2966)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3001..3020
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3050..3185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3112..3131
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3205..3224)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3206..3224)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3234..3251)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3234..3250)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3243..3265)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3392..3847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3479..3498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3689..3710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3873..3977
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3978..4838
                     /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(4196..4215)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5009..5597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5498..5517
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5751..5768
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5885..5906
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5921..5951
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5959..5975
                     /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     5964..5986
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5983..5999
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5983..5999
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6018..6038
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6020..6038
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6066..6292
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6293..6473
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6520..6645
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7138..7371
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7556..7600
                     /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    7867..7984
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8030..8540
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     8459..8477
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    8595..8619
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8691..8711
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8721..8747
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8751..8777
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8784..9497
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(8829..8850)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9090..9109)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9437..9458
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             9513..9569
                     /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             9576..10283
                     /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(9721..9739)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9989..10008
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             10290..10619
                     /codon_start=1
                     /product="degron consisting of residues 1-110 of human 
                     Geminin (Zielke and Edgar, 2015)"
                     /label=Gem1 (1-110)
                     /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
                     AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
                     KRRKAL"
     protein_bind    complement(10636..14)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggttgatat ccagcacagt ggcggccgct cgacaatcaa cctctggatt
       61 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
      121 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
      181 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
      241 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
      301 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
      361 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
      421 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
      481 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
      541 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
      601 cgagtcggat ctccctttgg gccgcctccc cgcctggaat tctgcagata tccggactag
      661 tgatctctcg aggttaacga attctaccgg gtaggggagg cgcttttccc aaggcagtct
      721 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct
      781 cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc
      841 gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg
      901 tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc
      961 gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc
     1021 gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc
     1081 aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag
     1141 cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcatcccg
     1201 ccaccatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     1261 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     1321 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     1381 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     1441 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     1501 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     1561 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     1621 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     1681 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     1741 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     1801 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     1861 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     1921 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     1981 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     2041 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     2101 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     2161 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     2221 caaaggaata gagtagatgc cgaccgaaca agagctgatt tcgagaacgc ctcagccagc
     2281 aactcgcgcg agcctagcaa ggcaaatgcg agagaacggc cttacgcttg gtggcacagt
     2341 tctcgtccac agttcgctaa gctcgctcgg ctgggtcgcg ggagggccgg tcgcagtgat
     2401 tcaggccctt ctggattgtg ttggtcccca gggcacgatt gtcatgccca cgcactcggg
     2461 tgatctgact gatcccgcag attggagatc gccgcccgtg cctgccgatt gggtgcagat
     2521 ccgtcgaggg cccggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     2581 actttttaaa agaaaagggg ggactggaag ggctagctca ctcccaacga agacaagatc
     2641 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     2701 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     2761 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     2821 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     2881 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     2941 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3001 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     3061 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3121 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3181 aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc
     3241 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3301 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3361 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     3421 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3481 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3541 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3601 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3661 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3721 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3781 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3841 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3901 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3961 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4021 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4081 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4141 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4201 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4261 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4321 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4381 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4441 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4501 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4561 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4621 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4681 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4741 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4801 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4861 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4921 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4981 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5041 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5101 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5161 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5221 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5281 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5341 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5401 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5461 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5521 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5581 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5641 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5701 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5761 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5821 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     5881 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     5941 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     6001 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
     6061 cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
     6121 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
     6181 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
     6241 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
     6301 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6361 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     6421 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
     6481 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
     6541 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
     6601 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
     6661 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
     6721 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
     6781 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
     6841 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
     6901 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
     6961 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
     7021 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     7081 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     7141 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     7201 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     7261 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     7321 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     7381 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     7441 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     7501 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     7561 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     7621 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     7681 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     7741 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     7801 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     7861 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     7921 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     7981 ttttatcgat cacgagacta gcctcgagaa gcttgatatc gaattccacg gggttggggt
     8041 tgcgcctttt ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt
     8101 ccgggaaacg cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg
     8161 tcacccggat cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc
     8221 taagtcggga aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac
     8281 gtctcactag taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc
     8341 gatgggctgt ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag
     8401 gggcggtgcg ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt
     8461 gttccgcatt ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa
     8521 tcaccgacct ctctccccag ggggatccac cggttcgtcg actagtccag tgtggtggaa
     8581 ttctgcagat atcaacaagt ttgtacaaaa aagcaggctt cgaattcatg gagcagcgcc
     8641 gcgtcaccga cttcttcgcg cgccgccgcc ccgggccccg gacacgtggc ccaaaaaaga
     8701 agagaaaggt aggagctcaa tacccatacg atgttcctga ctatgcgggc tatccctatg
     8761 acgtcccgga ctatgcaact agtatggtga gcaagggcga ggagctgttc accggggtgg
     8821 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     8881 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
     8941 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg cagtgcttcg
     9001 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     9061 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     9121 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     9181 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     9241 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     9301 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     9361 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     9421 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     9481 gcatggacga gctgtacctc gagggaagcg gagctactaa cttcagcctg ctgaagcagg
     9541 ctggagacgt ggaggagaac cctggacctc aattgatggt gagcaagggc gaggaggata
     9601 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     9661 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     9721 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     9781 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     9841 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     9901 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     9961 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
    10021 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
    10081 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
    10141 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
    10201 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
    10261 gcggcatgga cgagctgtac aaggtcgaga tgaatcccag tatgaagcag aaacaagaag
    10321 aaatcaaaga gaatataaag aatagttctg tcccaagaag aactctgaag atgattcagc
    10381 cttctgcatc tggatctctt gttggaagag aaaatgagct gtccgcaggc ttgtccaaaa
    10441 ggaaacatcg gaatgaccac ttaacatcta caacttccag ccctggggtt attgtcccag
    10501 aatctagtga aaataaaaat cttggaggag tcacccagga gtcatttgat cttatgatta
    10561 aagaaaatcc atcctctcag tattggaagg aagtggcaga aaaacggaga aaggcgctgt
    10621 aacaattggg atcctaccca gctttc
//
