pBOB-EF1-FastFUCCI-Puro

LOCUS       pBOB-EF1-FastFUCCI-Puro               12352 bp ds-DNA     circular SYN 12-MAR-2019
DEFINITION  3rd generation lentiviral vector encoding FUCCI reporters for cell 
            cycle analysis and puro for selection.
ACCESSION   .
VERSION     .
KEYWORDS    pBOB-EF1-FastFUCCI-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12352)
  AUTHORS   Koh SB, Mascalchi P, Rodriguez E, Lin Y, Jodrell DI, Richards FM, 
            Lyons SK
  TITLE     Quantitative FastFUCCI assay defines cell cycle dynamics at 
            single-cell level.
  JOURNAL   J Cell Sci. 2016 Nov 25. pii: jcs.195164.
  PUBMED    27888217
REFERENCE   2  (bases 1 to 12352)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 12, 2019 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12352
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        172..551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        553..751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     706..726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             769..949
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    996..1121
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1618..1851
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2332..2448
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2465..3643
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2696..3634
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3591..3611
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4648..4701
                     /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"
     promoter        5788..6287
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5823..5846)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6220..6239
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6331..6930
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6331..6350)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6827..6847
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7092..7680
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7145..7165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7563..7574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8091..8271
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8297..8314)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8303..8527
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8573..9001
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8660..8679)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8870..8891
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9010..9030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9015..9344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9195..9330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9257..9276
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9392..9439
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9458..9832
                     /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    9962..10083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9999..10018)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10053..10072
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10132..10148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10132..10148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10145..10167)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10156..10172
                     /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(10180..10210)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10225..10246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10363..10380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10534..11122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10614..10633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11293..12153)
                     /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     11916..11935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12154..12258)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12333..12352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
       61 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      121 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      541 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      601 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      661 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      721 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
      781 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      841 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      901 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      961 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
     1021 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     1081 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1141 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1201 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1261 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1321 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1381 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1441 agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag
     1501 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1561 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1621 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1681 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1741 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1801 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1861 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1921 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1981 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     2041 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     2101 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     2161 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     2221 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     2281 aggtggagag agagacagag acagatccat tcgattagtg aacggatcaa cttttaaaag
     2341 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2401 catacaaact aaagaattac aaaaacaaat tacaaaattc aaaattttat cttaattaag
     2461 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     2521 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     2581 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     2641 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     2701 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2761 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
     2821 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
     2881 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
     2941 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
     3001 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
     3061 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
     3121 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
     3181 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
     3241 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
     3301 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     3361 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     3421 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     3481 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     3541 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     3601 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgattaatta agaattggga
     3661 tccatggtgt ccgtgatcaa gcccgagatg aagatgcggt actacatgga cggcagcgtg
     3721 aacggccacg agttcaccat cgagggcgag ggcaccggca gaccttacga gggccaccag
     3781 gaaatgaccc tgagagtcac catggccgag ggcggaccca tgcccttcgc cttcgacctg
     3841 gtgtcccacg tgttctgcta cggccaccgg gtgttcacca agtaccccga ggaaatcccc
     3901 gactacttca agcaggcctt ccccgagggc ctgagctggg agcggagcct ggaattcgag
     3961 gatggcggca gcgcctctgt gtctgcccac atcagcctgc ggggcaacac cttctaccac
     4021 aagagcaagt tcaccggcgt gaacttcccc gccgacggcc ccatcatgca gaaccagagc
     4081 gtggactggg agcccagcac cgagaagatc accgccagcg acggcgtgct gaagggcgac
     4141 gtgaccatgt acctgaagct ggaaggcggc ggaaaccaca agtgccagat gaaaaccacc
     4201 tacaaggccg ccaaagaaat cctggaaatg cccggcgacc actacatcgg ccaccggctg
     4261 gtgcggaaaa ccgagggcaa catcaccgag caggtggagg atgccgtggc ccactacagc
     4321 atgctgccta gcctcggcgg ctaccctagc cattggaggc ccctggaacc ctctcctgcc
     4381 agacctgccc tgagagcccc tgcctccgcc acctccggca gcagaaagag agccagacct
     4441 cctgccgccc ctggcagaga tcaggccagg ccccctgccc gcagacggct ccggctgtcc
     4501 gtggatgagg tgtccagccc tagcaccccc gaggcccccg atatccctgc ctgtcccagc
     4561 cctggccaga aaatcaagaa gagcacacct gccgccggac agcctcctca cctgaccagc
     4621 gcccaggacc aggacaccat cgtcgacgag ggcaggggca gcctgctgac ctgcggcgac
     4681 gtggaggaga accccggccc cagatccatg gtgtccgtga tcaagcccga gatgaagatc
     4741 aagctgtgca tgcggggaac cgtgaacggc cacaacttcg tgatcgaggg cgagggcaag
     4801 ggaaacccct acgagggcac ccagatcctg gacctgaacg tgaccgaagg cgcccctctg
     4861 cccttcgcct acgacatcct gaccaccgtg ttccagtacg gcaaccgggc cttcaccaag
     4921 taccccgccg acatccagga ctacttcaag cagaccttcc ccgagggcta ccactgggag
     4981 cggagcatga cctacgagga ccagggcatc tgcaccgcca cctccaacat cagcatgagg
     5041 ggcgactgct ttttctacga catcagattc gacggcacca acttcccccc caacggcccc
     5101 gtgatgcaga aaaagaccct gaagtgggag cccagcaccg agaagatgta cgtggaggac
     5161 ggcgtgctga agggcgacgt gaacatgcgg ctgctgctgg aaggcggcgg acactacaga
     5221 tgcgacttca agaccaccta caaggccaag aaagaagtgc ggctgcccga cgcccacaag
     5281 atcgaccacc ggatcgagat cctgaagcac gacaaggact acaacaaagt gaagctgtac
     5341 gagaacgccg tggccaggta cagcatgctg cccagccagg ccaagggcta ccccagccac
     5401 tggcggcctc tggaaatgaa ccccagcatg aagcagaagc aggaagagat caaagagaac
     5461 atcaagaaca gcagcgtgcc caggcggacc ctgaagatga tccagcccag cgccagcgga
     5521 tctctggtgg gccgggagaa tgagctgtct gccggcctga gcaagcggaa gcaccggaac
     5581 gaccacctga ccagcaccac ctctagccca ggcgtgatcg tgcccgagag cagcgagaac
     5641 aagaacctgg gcggcgtgac ccaagaaagc ttcgacctga tgatcaaaga aaaccccagc
     5701 agccagtact ggaaagaggt ggccgagaag cggcggaagg ccctgtaggt cgagcaattc
     5761 gcggccgcgc tagctcgaaa ttctaccggg taggggaggc gcttttccca aggcagtctg
     5821 gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc
     5881 gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg
     5941 cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt
     6001 gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg
     6061 ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg
     6121 ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca
     6181 ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc
     6241 gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc tgcatccatc
     6301 tagatctcga tcgagcagct gaagcttacc atgaccgagt acaagcccac ggtgcgcctc
     6361 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     6421 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     6481 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     6541 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     6601 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     6661 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     6721 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     6781 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     6841 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     6901 tggtgcatga cccgcaagcc cggtgcctga gctagctgca ggtcgagcat gcatctaggg
     6961 cggccaattc cgccgatcct ggaaaaacat ggagcaatca caagtagcaa tacagcagct
     7021 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc
     7081 acacctcaga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     7141 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     7201 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     7261 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     7321 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     7381 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     7441 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     7501 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     7561 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     7621 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     7681 ctggaattaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     7741 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     7801 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
     7861 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
     7921 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
     7981 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
     8041 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
     8101 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     8161 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     8221 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggccgctt
     8281 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     8341 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     8401 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     8461 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     8521 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     8581 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     8641 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     8701 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     8761 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     8821 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     8881 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     8941 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     9001 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     9061 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     9121 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     9181 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     9241 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     9301 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     9361 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     9421 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     9481 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     9541 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     9601 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     9661 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     9721 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     9781 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     9841 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     9901 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     9961 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
    10021 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
    10081 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
    10141 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    10201 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    10261 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    10321 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    10381 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    10441 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    10501 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    10561 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    10621 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    10681 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    10741 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    10801 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    10861 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    10921 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    10981 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    11041 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    11101 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    11161 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    11221 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    11281 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    11341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    11401 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    11461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    11521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    11581 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    11641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    11701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    11761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    11821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    11881 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    11941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    12001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    12061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    12121 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    12181 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    12241 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
    12301 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cg
//