LOCUS       sequence-172473-        9062 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Constitutively express DsRed and conditionally express Puro and 
            EGFP. Used to test transfection efficiency and enrich genetically 
            modified cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9062)
  AUTHORS   Ren C, Xu K, Liu Z, Shen J, Han F, Chen Z, Zhang Z
  TITLE     Dual-reporter surrogate systems for efficient enrichment of 
            genetically modified cells.
  JOURNAL   Cell Mol Life Sci. 2015 Jul;72(14):2763-72. doi: 
            10.1007/s00018-015-1874-6. Epub 2015 Mar 1.
  PUBMED    25725802
REFERENCE   2  (bases 1 to 9062)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1007/s00018-015-1874-6"; journalName: "Cell Mol Life Sci"; date:
            "2015-07"; volume: "72"; issue: "14"; pages: "2763-72"
COMMENT     Sequence Label: pNHEJ-RPG
FEATURES             Location/Qualifiers
     source          1..9062
                     /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"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2340..2374
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        2753..3140
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3141..3344
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3295..3315
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3378..3387
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3384..3386,3387..3389,3390..4064)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(3567..3587)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3785..3804
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3979..4002
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4068..4087)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4133..4188
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4187..4206)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        4561..4940
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4942..5219
                     /label=chicken beta-actin promoter
     intron          5220..6236
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     6160..6182
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     6244..6263
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      6292..6301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6318..6911
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6811..6831
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             6936..6989
                     /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             join(6990..6992,6993..7706)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIDDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(7032..7053)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7293..7312)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7640..7661
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7824..7945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7861..7880)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7915..7934
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8295..8357
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8423..8442)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8424..8442)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8449..8466)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8449..8465)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8458..8480)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8607..9035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8694..8713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8904..8925
                     /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 cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtacctc gcgcgacttg
     2221 gtttgccatt ctttagcgcg cgtcgcgtca cacagcttgg ccacaatgtg gtttttgtca
     2281 aacgaagatt ctatgacgtg tttaaagttt aggtcgagta aagcgcaaat cttttttaac
     2341 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
     2401 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
     2461 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
     2521 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
     2581 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
     2641 tagatatcgt gactaatata taataaaatg ggtagttctt tagacgatga gcatatcctc
     2701 tctgctcttc tgcaaagcga tgacgagctt gttggctagc ggtggcggcc tcgacattga
     2761 ttattgacta gagcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     2821 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     2881 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     2941 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     3001 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     3061 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     3121 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     3181 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     3241 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     3301 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     3361 agatccgcta gccggtcgcc accatggtgc gctcctccaa gaacgtcatc aaggagttca
     3421 tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag atcgagggcg
     3481 agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg accaagggcg
     3541 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt
     3601 acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca
     3661 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     3721 ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg
     3781 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     3841 cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc
     3901 actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct
     3961 actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     4021 agcagtacga gcgcaccgag ggccgccacc acctgttcct gtaaggatct ttttccctct
     4081 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     4141 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     4201 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     4261 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     4321 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     4381 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     4441 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     4501 ctcttatgga gatccctcga cctgcagccc aagcttatcg atactagcgg tggcggcctc
     4561 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     4621 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4681 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4741 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4801 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4861 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4921 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     4981 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     5041 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
     5101 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
     5161 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
     5221 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
     5281 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
     5341 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
     5401 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
     5461 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
     5521 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     5581 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     5641 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     5701 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     5761 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     5821 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     5881 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     5941 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     6001 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     6061 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     6121 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     6181 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     6241 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc accatggcgg
     6301 ccgcgatatc ggatcccacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     6361 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     6421 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     6481 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     6541 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     6601 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     6661 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     6721 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     6781 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     6841 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     6901 agcccggtgc cgctcgagct accggtggca gtggagaggg cagaggaagt ctgctaacat
     6961 gcggtgacgt cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg
     7021 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     7081 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     7141 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     7201 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     7261 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     7321 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     7381 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     7441 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     7501 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcgacgacg
     7561 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     7621 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     7681 tcggcatgga cgagctgtac aagtaatcta gataactgat cataatcagc cataccacat
     7741 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     7801 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     7861 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7921 tgtccaaact catcaatgta tcttagtcga gttaattaac gagagcataa tattgatatg
     7981 tgccaaagtt gtttctgact gactaataag tataatttgt ttctattatg tataggttaa
     8041 gctaattact tattttataa tacaacatga ctgtttttaa agtacaaaat aagtttattt
     8101 ttgtaaaaga gagaatgttt aaaagttttg ttactttata gaagaaattt tgagtttttg
     8161 ttttttttta ataaataaat aaacataaat aaattgtttg ttgaatttat tattagtatg
     8221 taagtgtaaa tataataaaa cttaatatct attcaaatta ataaataaac ctcgatatac
     8281 agaccgataa aacacatgcg tcaattttac gcatgattat ctttaacgta cgtcacaata
     8341 tgattatctt tctagggtta aataatagtt tctaattttt ttattattca gcctgctgtc
     8401 gtgaataccg agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
     8461 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     8521 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     8581 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     8641 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     8701 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     8761 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     8821 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     8881 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     8941 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     9001 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     9061 tt
//
