LOCUS       sequence-476234-       12786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible firefly luciferase and mCherry; constitutive far-red 
            iRFP720.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12786)
  AUTHORS   Walton BL, Shattuck-Brandt R, Hamann CA, Tung VW, Colazo JM, Brand 
            DD, Hasty KA, Duvall CL, Brunger JM
  TITLE     A programmable arthritis-specific receptor for guided articular 
            cartilage regenerative medicine.
  JOURNAL   Osteoarthritis Cartilage. 2025 Feb;33(2):231-240. doi: 
            10.1016/j.joca.2024.12.002. Epub 2024 Dec 18.
  PUBMED    39706287
REFERENCE   2  (bases 1 to 12786)
  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"; doi: 
            "10.1016/j.joca.2024.12.002"; journalName: "Osteoarthritis 
            Cartilage"; date: "2025-02"; volume: "33"; issue: "2"; pages: 
            "231-240"
COMMENT     Sequence Label: pHR TRE_firefly Luciferase_2A_mCherry-PGK_iRFP720
FEATURES             Location/Qualifiers
     source          1..12786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    28..145
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        224..591
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    232..250
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    268..286
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    304..322
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    340..358
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    376..394
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    412..430
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    448..466
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      630..639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2289..2345
                     /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             2349..3059
                     /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(2494..2512)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2762..2781
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        3068..3567
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3103..3126)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3500..3519
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3568..3577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3574..4524
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     misc_feature    4628..5216
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4681..4701)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5099..5110)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5219..5235)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5220..5236)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5326..5559
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(5584..5602)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5585..5602)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5892..5911)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6125..6144)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6244..6266
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6304..6322)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6390..6494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6495..6563,6564..7355)
                     /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(6713..6732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7526..8114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8015..8034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8268..8285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8355..8375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8360..8689
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8540..8675
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          9823..9888
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             10018..10038
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10463..10597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10500..10519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10554..10573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10765..11398
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    11445..11570
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12067..12300
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12485..12529
                     /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"
ORIGIN
        1 tcgccaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
       61 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
      121 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg
      181 gatcgataag cttgatatcg aattcccgtg gaattatcac ctcgagttta ctccctatca
      241 gtgatagaga acgtatgaag agtttactcc ctatcagtga tagagaacgt atgcagactt
      301 tactccctat cagtgataga gaacgtataa ggagtttact ccctatcagt gatagagaac
      361 gtatgaccag tttactccct atcagtgata gagaacgtat ctacagttta ctccctatca
      421 gtgatagaga acgtatatcc agtttactcc ctatcagtga tagagaacgt ataagctttg
      481 cttatgtaaa ccagggcgcc tataaaagag tgctgatttt ttgagtaaac ttcaattcca
      541 caacactttt gtcttatacc aactttccgt accacttcct accctcgtaa agaattcccg
      601 tatacaccgg tgcaattctc acgcgtgccg ccaccatgga agatgccaag aacatcaaga
      661 aaggccctgc ccccttctac cccctggaag atggcacagc cggcgagcag ctgcacaagg
      721 ccatgaagag atacgccctg gtgcccggca ccatcgcctt caccgacgcc cacatcgagg
      781 tggacatcac ctacgccgag tatttcgaga tgagcgtgcg gctggccgag gccatgaaac
      841 gctacggcct gaacaccaac caccggatcg tggtgtgcag cgagaacagc ctgcagttct
      901 tcatgcccgt gctgggcgcc ctgttcatcg gcgtggccgt ggcccctgcc aacgacatct
      961 acaacgagcg ggagctgctg aacagcatgg gcatcagcca gcccaccgtg gtgttcgtga
     1021 gcaagaaggg cctgcagaaa atcctgaacg tgcagaagaa gctgcccatc atccagaaaa
     1081 tcatcatcat ggacagcaag accgactacc agggcttcca gagcatgtac accttcgtga
     1141 ccagccacct gccccctggc ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg
     1201 acaagaccat cgccctgatc atgaacagca gcggcagcac cggcctgcct aaaggcgtgg
     1261 ccctgcctca ccggaccgcc tgcgtgcggt tcagccacgc ccgggacccc atcttcggca
     1321 accagatcat ccccgacacc gccatcctga gcgtggtgcc cttccaccac ggcttcggca
     1381 tgttcaccac cctgggctac ctgatctgcg gcttccgggt ggtgctgatg taccggttcg
     1441 aggaagagct gttcctgcgg agcctgcagg actacaagat ccagagcgcc ctgctggtgc
     1501 ccaccctgtt cagctttttc gccaagagca ccctgatcga caagtacgac ctgagcaacc
     1561 tgcacgagat cgccagcggc ggagcccccc tgtccaaaga agtgggcgaa gccgtcgcca
     1621 agcggttcca cctgcccggc atccggcagg gctatggcct gaccgagacc acaagcgcca
     1681 ttctgatcac ccccgagggc gacgacaagc ctggcgccgt gggcaaggtg gtgcctttct
     1741 tcgaggccaa ggtggtggac ctggacaccg gcaagaccct gggcgtgaac cagcggggcg
     1801 agctgtgcgt gaggggcccc atgatcatga gcggctacgt gaacaacccc gaggccacca
     1861 acgccctgat tgacaaggac ggctggctgc acagcggcga catcgcctac tgggacgagg
     1921 acgagcactt cttcatcgtg gaccggctga agagcctgat caagtacaag ggctaccagg
     1981 tggccccagc cgagctggaa agcatcctgc tgcagcaccc caacatcttc gatgccgggg
     2041 tggccggact gcccgacgac gatgccggcg agctgcctgc cgccgtggtg gtgctggaac
     2101 acggcaaaac catgaccgag aaagaaatcg tggactacgt ggccagccag gtgaccaccg
     2161 ccaagaaact gagaggcggc gtggtgtttg tggacgaggt gcccaagggc ctgacaggca
     2221 agctggacgc ccggaagatc cgggagatcc tgatcaaggc caagaagggc ggcaagagag
     2281 ccaagagagc cacaaacttc tctctgctaa agcaagcagg tgatgttgaa gaaaaccccg
     2341 ggcctcatat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     2401 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     2461 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     2521 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     2581 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     2641 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     2701 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     2761 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     2821 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     2881 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     2941 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     3001 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag
     3061 cggccgcggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
     3121 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
     3181 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
     3241 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
     3301 gtagcacgtc tcactagcct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
     3361 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
     3421 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
     3481 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     3541 tcctcttcct catctccggg cctttcggcc accatggcgg aaggatccgt cgccaggcag
     3601 cctgacctct tgacctgcga cgatgagccg atccatatcc ccggtgccat ccaaccgcat
     3661 ggactgctgc tcgccctcgc cgccgacatg acgatcgttg ccggcagcga caaccttccc
     3721 gaactcaccg gactggcgat cggcgccctg atcggccgct ctgcggccga tgtcttcgac
     3781 tcggagacgc acaaccgtct gacgatcgcc ttggccgagc ccggggcggc cgtcggagca
     3841 ccgatcactg tcggcttcac gatgcgaaag gacgcaggct tcatcggctc ctggcatcgc
     3901 catgatcagc tcatcttcct cgagctcgag cctccccagc gggacgtcgc cgagccgcag
     3961 gcgttcttcc gccgcaccaa cagcgccatc cgccgcctgc aggccgccga aaccttggaa
     4021 agcgcctgcg ccgccgcggc gcaagaggtg cggaagatta ccggcttcga tcgggtgatg
     4081 atctatcgct tcgcctccga cttcagcggg tccgtgatcg cagaggatcg gtgcgccgag
     4141 gtcgagtcaa aactaggcct gcactatcct gcctcattca tcccggcgca ggcccgtcgg
     4201 ctctatacca tcaacccggt acggatcatt cccgatatca attatcggcc ggtgccggtc
     4261 accccagacc tcaatccggt caccgggcgg ccgattgatc ttagcttcgc catcctgcgc
     4321 agcgtctcgc ccaaccatct ggagttcatg cgcaacatag gcatgcacgg cacgatgtcg
     4381 atctcgattt tgcgcggcga gcgactgtgg ggattgatcg tttgccatca ccgaacgccg
     4441 tactacgtcg atctcgatgg ccgccaagcc tgcgagctag tcgcccaggt tctggcctgg
     4501 cagatcggcg tgatggaaga gtaattaatt aaggccgcaa ctcccacctg caacatgcgt
     4561 gactgactga ggccgcgact ctagagtcga cctgcaggca tgcaagcttg atatcaagct
     4621 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     4681 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     4741 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     4801 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     4861 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     4921 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     4981 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5041 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5101 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5161 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     5221 ataccgtcga cctcgaggga attaattcga gctcggtacc tttaagacca atgacttaca
     5281 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     5341 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     5401 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     5461 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     5521 ccctcagacc cttttagtca gtgtggaaaa tctctagcag catctagaat taattccgtg
     5581 tattctatag tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta
     5641 gccgcgttct aacgacaata tgtacaagcc taattgtgta gcatctggct tactgaagca
     5701 gaccctatca tctctctcgt aaactgccgt cagagtcggt ttggttggac gaaccttctg
     5761 agtttctggt aacgccgtcc cgcacccgga aatggtcagc gaaccaatca gcagggtcat
     5821 cgctagccag atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     5881 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     5941 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     6001 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     6061 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgaa tggtgcactc
     6121 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     6181 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6241 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6301 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6361 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6481 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6541 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6601 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6661 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6721 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6781 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6841 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6901 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6961 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7021 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     7081 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7141 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7201 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7261 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7321 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7381 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7441 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8161 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8221 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8281 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     8341 attaatgcag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     8401 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     8461 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     8521 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     8581 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     8641 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttggaca
     8701 caagacaggc ttgcgagata tgtttgagaa taccacttta tcccgcgtca gggagaggca
     8761 gtgcgtaaaa agacgcggac tcatgtgaaa tactggtttt tagtgcgcca gatctctata
     8821 atctcgcgca acctattttc ccctcgaaca ctttttaagc cgtagataaa caggctggga
     8881 cacttcacat gagcgaaaaa tacatcgtca cctgggacat gttgcagatc catgcacgta
     8941 aactcgcaag ccgactgatg ccttctgaac aatggaaagg cattattgcc gtaagccgtg
     9001 gcggtctgta ccgggtgcgt tactggcgcg tgaactgggt attcgtcatg tcgataccgt
     9061 ttgtatttcc agctacgatc acgacaacca gcgcgagctt aaagtgctga aacgcgcaga
     9121 aggcgatggc gaaggcttca tcgttattga tgacctggtg gataccggtg gtactgcggt
     9181 tgcgattcgt gaaatgtatc caaaagcgca ctttgtcacc atcttcgcaa aaccggctgg
     9241 tcgtccgctg gttgatgact atgttgttga tatcccgcaa gatacctgga ttgaacagcc
     9301 gtgggatatg ggcgtcgtat tcgtcccgcc aatctccggt cgctaatctt ttcaacgcct
     9361 ggcactgccg ggcgttgttc tttttaactt caggcgggtt acaatagttt ccagtaagta
     9421 ttctggaggc tgcatccatg acacaggcaa acctgagcga aaccctgttc aaaccccgct
     9481 ttaaacatcc tgaaacctcg acgctagtcc gccgctttaa tcacggcgca caaccgcctg
     9541 tgcagtcggc ccttgatggt aaaaccatcc ctcactggta tcgcatgatt aaccgtctga
     9601 tgtggatctg gcgcggcatt gacccacgcg aaatcctcga cgtccaggca cgtattgtga
     9661 tgagcgatgc cgaacgtacc gacgatgatt tatacgatac ggtgattggc taccgtggcg
     9721 gcaactggat ttatgagtgg gccccggatc tttgtgaagg aaccttactt ctgtggtgtg
     9781 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
     9841 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
     9901 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
     9961 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
    10021 aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg
    10081 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
    10141 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
    10201 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
    10261 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
    10321 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
    10381 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
    10441 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
    10501 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    10561 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
    10621 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
    10681 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta
    10741 aatatgtact acaaacttag tagttggaag ggctaattca ctcccaaaga agacaagata
    10801 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac
    10861 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
    10921 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga
    10981 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc
    11041 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc
    11101 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga
    11161 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg
    11221 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
    11281 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
    11341 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
    11401 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
    11461 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
    11521 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
    11581 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
    11641 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
    11701 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
    11761 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
    11821 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
    11881 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg
    11941 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
    12001 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
    12061 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
    12121 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
    12181 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
    12241 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
    12301 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
    12361 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
    12421 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
    12481 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
    12541 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
    12601 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
    12661 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
    12721 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
    12781 acggta
//
