LOCUS       sequence-171531-       10875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lenti plasmid for generating CIAR expressing stable cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10875)
  AUTHORS   Rose JC, Huang PS, Camp ND, Ye J, Leidal AM, Goreshnik I, Trevillian
            BM, Dickinson MS, Cunningham-Bryant D, Debnath J, Baker D, 
            Wolf-Yadlin A, Maly DJ
  TITLE     A computationally engineered RAS rheostat reveals RAS-ERK signaling 
            dynamics.
  JOURNAL   Nat Chem Biol. 2016 Nov 21. doi: 10.1038/nchembio.2244.
  PUBMED    27870838
REFERENCE   2  (bases 1 to 10875)
  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: "Nat Chem 
            Biol. 2016 Nov 21. doi: 10.1038/nchembio.2244."
COMMENT     Sequence Label: GFP-P2A-CIAR_pLenti_CMV_Puro
FEATURES             Location/Qualifiers
     source          1..10875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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     2117..2139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2136..2152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2171..2191
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2219..2445
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2446..2626
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2673..2798
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3291..3524
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3709..3753
                     /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    4020..4136
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        4159..4462
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4463..4666
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4617..4637
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4663..4687
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4756..4765
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4762..4764,4765..4767,4768..5478)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4762..4764,4765..4767,4768..5478)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4807..4828)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5068..5087)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5415..5436
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5488..5544
                     /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"
     misc_feature    7774..8362
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7827..7847)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8245..8256)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        8376..8875
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8411..8434)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8808..8827
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8896..9495
                     /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(8896..8915)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9392..9412
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             9633..9866
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9938..10059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9975..9994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10029..10048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10078..10213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10233..10252)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10234..10252)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10262..10279)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10262..10278)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10271..10293)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10420..10848
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10507..10526)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10717..10738
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     2221 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     2281 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     2341 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     2401 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     2461 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2521 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2581 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     2641 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     2701 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     2761 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     2821 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     2881 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     2941 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     3001 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     3061 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     3121 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     3181 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     3241 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     3301 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     3361 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     3421 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     3481 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     3541 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     3601 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     3661 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     3721 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     3781 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     3841 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     3901 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     3961 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact
     4021 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     4081 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaattcaa aattttatcg
     4141 ataagcttgg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     4201 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     4261 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     4321 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     4381 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     4441 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     4501 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     4561 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     4621 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     4681 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgactcta
     4741 gaggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     4801 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     4861 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     4921 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     4981 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     5041 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     5101 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5161 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     5221 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     5281 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     5341 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     5401 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     5461 atggacgagc tgtacaaggg aagcggagct actaacttca gcctgctgaa gcaggctgga
     5521 gacgtggagg agaaccctgg acctgcacca ggaagtatgg ctcctcctaa tctgtgggcg
     5581 gcacagcgtt atgggcgtga actgcggcgt atggccgatg agggtgaggg ttccttcaag
     5641 ggtagcggaa ctgggtctgg gactgggagt ggcaccggtt cagggacagg agatgtgtat
     5701 cgctttgctg agcctgactc cgaagagaac attatcttcg aagagaatat gcagcctaaa
     5761 gctggtattc cgattatcaa agccgggacc gtgattaagc tgatcgagcg tctgacatat
     5821 cacatgtacg ccgaccccaa ctttgttcgc accttcctga ccacatatcg ttccttttgc
     5881 aagccacagg aactgctgtc tctgattatc gagcggttcg aaattccaga accagagcca
     5941 accgaggcag atcgtattgc gatcgaaaac ggcgaccagc ctctgagcgc ggagctgaaa
     6001 cgttttcgga aggaatacat ccagccggtc cagctgcggg ttctgaatgt ctgtcgccat
     6061 tgggtggagc accattttta tgatttcgaa cgggacgctt acctgctgca gcgcatggaa
     6121 gagttcattg gtacagtccg cgggaaagcc atgaaaaagt gggtggaatc cattactaag
     6181 attatccagc gcaaaaagat cgcccgtgat aacgggccag gacacaatat cacatttcag
     6241 agctccccgc ccactgtgga gtggcacatt tctcgtcccg gtcatatcga aaccttcgat
     6301 ctgctgacac tgcatccaat tgagatcgct cgtcagctga cactgctgga atctgacctg
     6361 tatcgtgcag tgcagccaag cgagctggtg ggctccgttt ggaccaaaga ggataaggaa
     6421 attaacagtc caaatctgct gaaaatgatc cgccacacta ccaacctgac tctgtggttt
     6481 gaaaagtgca ttgttgaaac cgagaatctg gaagagcgtg tcgcggtggt tagccggatt
     6541 atcgagatcc tgcaggtgtt tcaggaactg aacaatttca acggcgttct ggaggtcgtg
     6601 tccgcaatga acagcagccc tgtgtatcgc ctggaccata ccttcgaaca gattccgagc
     6661 cgtcagaaaa agatcctgga agaggcgcac gagctgtccg aagatcatta taaaaagtac
     6721 ctggcaaaac tgcgctctat taacccacct tgcgtgccgt ttttcggtat ttacctgact
     6781 aacatcctga agaccgaaga gggaaatccg gaggttctga aacgtcacgg caaggaactg
     6841 atcaactttt caaaacgccg taaggtcgcc gagattctcg gtgaaatcca gcagtatcag
     6901 aatcagcctt actgtctgcg tgtggagagc gacattaaac ggtttttcga aaacctgaat
     6961 ccgatgggga acagtatgga aaaagagttt accgattacc tgttcaataa gtcactggaa
     7021 atcgagcctg gctctggttc acagagcaac cgtgaactgg ttgtcgattt tctgtcctat
     7081 aaactgtctc agaagggcta ctcctggtct cagttcagcg acgtcgaaga gaatcggaca
     7141 gaggctccgg aaggtactga atctgagatg gaaaccccca gtgccattaa cgggaatcca
     7201 tcatggcacc tggcagatag ccccgctgtt aatggagcca ccggacattc gagctccctg
     7261 gacgcccgcg aggttatccc aatggccgca gtcaaacagg cgctgcgtga agcaggagat
     7321 gagtttgaac tgcgctatcg gcgcgcgttc agtgacctga cctcacagct gcacattaca
     7381 cccggcactg cataccagag ttttgagcag gtggttaacg aactgttccg cgacggagtg
     7441 aattggggcc gtatcgttgc gtttttcagt tttggcggtg cactgtgtgt cgagtcagtg
     7501 gataaagaaa tgcaggtcct ggtgtctcgt attgcagctt ggatggcaac ttatctgaac
     7561 gatcatctgg agccgtggat tcaggaaaat gggggatggg acaccttcgt ggaactgtac
     7621 ggcaacaatg cggcggctga gagtcgtaag gggcaggagc ggttcaatcg ttggtttctg
     7681 cagcacaagc tgcggaagct gaaccctcct gatgagagtg gccccggctg catgagctgc
     7741 aagtgtgtgc tctcctgaag cggccgcgtc gacaatcaac ctctggatta caaaatttgt
     7801 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     7861 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     7921 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     7981 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     8041 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     8101 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     8161 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     8221 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     8281 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     8341 tccctttggg ccgcctcccc gcctggaatt ctaccgggta ggggaggcgc ttttcccaag
     8401 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     8461 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     8521 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     8581 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     8641 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     8701 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     8761 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     8821 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     8881 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     8941 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     9001 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     9061 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     9121 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     9181 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     9241 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     9301 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     9361 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     9421 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     9481 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
     9541 ccccatgcat ctcgagggcc cggtaccttt aagaccaatg acttacaagg cagctgtaga
     9601 tcttagccac tttttaaaag aaaagggggg actggaaggg ctagctcact cccaacgaag
     9661 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     9721 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     9781 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     9841 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     9901 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
     9961 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
    10021 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
    10081 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
    10141 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
    10201 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
    10261 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
    10321 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
    10381 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
    10441 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
    10501 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
    10561 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
    10621 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
    10681 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
    10741 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
    10801 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
    10861 taacgcttac aattt
//
