LOCUS       sequence-417519-       11710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible lentiviral plasmid expressing NR3C1-P2A-mCherry, with 
            ORF specific barcode close to 3'LTR site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11710)
  AUTHORS   Obradovic A, Ager C, Turunen M, Nirschl T, Khosravi-Maharlooei M, 
            Iuga A, Jackson CM, Yegnasubramanian S, Tomassoni L, Fernandez EC, 
            McCann P, Rogava M, DeMarzo AM, Kochel CM, Allaf M, Bivalacqua T, 
            Lim M, Realubit R, Karan C, Drake CG, Califano A
  TITLE     Systematic elucidation and pharmacological targeting of 
            tumor-infiltrating regulatory T cell master regulators.
  JOURNAL   Cancer Cell. 2023 May 8;41(5):933-949.e11. doi: 
            10.1016/j.ccell.2023.04.003. Epub 2023 Apr 27.
  PUBMED    37116491
REFERENCE   2  (bases 1 to 11710)
  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.ccell.2023.04.003"; journalName: "Cancer Cell"; date: 
            "2023-05-8- 8"; volume: "41"; issue: "5"; pages: "933-949.e11"
COMMENT     Sequence Label: Tet-O-FUW-NR3C1-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..11710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(138..336)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        338..717
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     889..908
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        983..1087
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1088..1156,1157..1948)
                     /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(1306..1325)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2119..2707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2608..2627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2861..2878
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2995..3016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3031..3036,3037..3054,3055..3061)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3069..3085
                     /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     3074..3096
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3093..3109
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3093..3109
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3158..3279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3169..3188)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3223..3242
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3409..3783)
                     /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"
     promoter        complement(3802..3849)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3897..4226)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3911..4046
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3965..3984)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4211..4231
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4240..4668)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4350..4371)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4562..4581
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4714..4938
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4927..4944
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             4970..5150
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     5701..5717
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5702..5718
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5721..6309
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5827..5838
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6236..6256
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6332..7039)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(6610..6629)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6879..6897
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(7049..7105)
                     /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             complement(7130..9460)
                     /codon_start=1
                     /gene="Homo sapiens NR3C1"
                     /product="human glucocorticoid receptor, isoform alpha"
                     /label=glucocorticoid receptor
                     /note="acts as a transcription factor that binds to 
                     glucocorticoid response elements, and as a modulator of 
                     other transcription factors"
                     /translation="MDSKESLTPGREENPSSVLAQERGDVMDFYKTLRGGATVKVSASS
                     PSLAVASQSDSKQRRLLVDFPKGSVSNAQQPDLSKAVSLSMGLYMGETETKVMGNDLGF
                     PQQGQISLSSGETDLKLLEESIANLNRSTSVPENPKSSASTAVSAAPTEKEFPKTHSDV
                     SSEQQHLKGQTGTNGGNVKLYTTDQSTFDILQDLEFSSGSPGKETNESPWRSDLLIDEN
                     CLLSPLAGEDDSFLLEGNSNEDCKPLILPDTKPKIKDNGDLVLSSPSNVTLPQVKTEKE
                     DFIELCTPGVIKQEKLGTVYCQASFPGANIIGNKMSAISVHGVSTSGGQMYHYDMNTAS
                     LSQQQDQKPIFNVIPPIPVGSENWNRCQGSGDDNLTSLGTLNFPGRTVFSNGYSSPSMR
                     PDVSSPPSSSSTATTGPPPKLCLVCSDEASGCHYGVLTCGSCKVFFKRAVEGQHNYLCA
                     GRNDCIIDKIRRKNCPACRYRKCLQAGMNLEARKTKKKIKGIQQATTGVSQETSENPGN
                     KTIVPATLPQLTPTLVSLLEVIEPEVLYAGYDSSVPDSTWRIMTTLNMLGGRQVIAAVK
                     WAKAIPGFRNLHLDDQMTLLQYSWMFLMAFALGWRSYRQSSANLLCFAPDLIINEQRMT
                     LPCMYDQCKHMLYVSSELHRLQVSYEEYLCMKTLLLLSSVPKDGLKSQELFDEIRMTYI
                     KELGKAIVKREGNSSQNWQRFYQLTKLLDSMHEVVENLLNYCFQTFLDKTMSIEFPEML
                     AEIITNQIPKYSNGNIKKLLFHQK"
     regulatory      9457..9466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(9546..9570)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(9567..9605)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    9638..9656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9680..9698
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9722..9740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9764..9782
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     gap             9801..9900
                     /estimated_length=100
     protein_bind    9906..9924
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    9981..10098
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(10396..10440)
                     /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    10625..10858
                     /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    11351..11476
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             11523..11703
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag
       61 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
      121 agtacaggca aaacgcgctg cttatataga cctcccaccg tacacgccta ccgcccattt
      181 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
      241 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
      301 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
      361 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
      421 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
      481 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
      541 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
      601 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
      661 tatgaactaa tgaccccgta attgattact attaataact agtcaataat caatgtcaac
      721 gcgtatatct ggcccgtaca tcgcgaagca gcgcaaaacg cctaacccta agcagattct
      781 tcatgcaatt gtcggtcaag ccttgccttg ttgtagctta aattttgctc gcgcactact
      841 cagcgacctc caacacacaa gcagggagca gatactggct taactatgcg gcatcagagc
      901 agattgtact gagagtgcac cataggggat cgggagatct cccgatccgt cgacgtcagg
      961 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1021 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1081 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1141 ccttcctgtt tttgctcacc cagaaacgct ggtaaaagta aaagatgctg aagatcagtt
     1201 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1261 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1321 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1381 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1441 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1501 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1561 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1621 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1681 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1741 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1801 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1861 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1921 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1981 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2041 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2101 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2161 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2221 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2281 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2341 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2401 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2461 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2521 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2581 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2641 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2701 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2761 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2821 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2881 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2941 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     3001 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     3061 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     3121 caagctctag ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga
     3181 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3241 gctttatttg taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag
     3301 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     3361 caacctttca tagaaggcgg cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc
     3421 ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca
     3481 cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac
     3541 gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt
     3601 gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac
     3661 cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa
     3721 ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc
     3781 catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg ccgatgatta
     3841 attgtcaaca cgtgctgatc agatccgaaa atggatatac aagctcccgg gagctttttg
     3901 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag
     3961 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     4021 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     4081 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     4141 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     4201 gactttccac accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt
     4261 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4321 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4381 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4441 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4501 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4561 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4621 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc
     4681 tagagcccca gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat
     4741 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     4801 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     4861 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     4921 ggcaactaga aggcacagtc gaggctgatc agcgggttta aacgggccct gctagagatt
     4981 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
     5041 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
     5101 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc agtacagtcc
     5161 ggatgcagct ctcgggccat gtgatgaaat gctaggcggc tgtcaaacct ccactctaat
     5221 acttctctct ccgggtcatc catcccatgc aggctcacag ggtgtaacaa gcgggtgttc
     5281 tctccttcat tggcttcttc taccttctct tgctcaactg gtactagttc tgctagatgg
     5341 aactgagtac actgcctaat atcacgtcct agccttgtag caccatccaa aggtcagtgg
     5401 atatctgatc cctggccctg gtgtgtagtt ctgccaatca gggaagtagc cttgtgtgtg
     5461 gtagatccac agatcaagga tatcttgtct tcgttgggag tgaattagcc cttccagtcc
     5521 ccccttttct tttaaaaagt ggctaagatc tacagctgcc ttgtaagtca ttggtcttaa
     5581 aggtacctga ggtgtgactg gaaaacccac ctcctcctcc tcttgtgctt ctagccaggc
     5641 acaatcagca ttggtagctg ctgtattgct acttgtgatt gctccatgtt tttctaggtc
     5701 tcgaggtcga cggtatcgat gcggggaagc ggcccaaagg gagatccgac tcgtctgagg
     5761 gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg aaggaaggtc
     5821 cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc
     5881 aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca cggaattgtc
     5941 agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga tgagttccgc
     6001 cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg tggtggcaat
     6061 gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac gttgcctgac
     6121 aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg aggagaaaat
     6181 gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt atccacatag
     6241 cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt tgtaatccag
     6301 aggttgattg tcgacttaac gcgttctaga ttcacttgta cagctcgtcc atgccgccgg
     6361 tggagtggcg gccctcggcg cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg
     6421 aggtgatgtc caacttgatg ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg
     6481 ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct
     6541 gcttgatctc gcccttcagg gcgccgtcct cggggtacat ccgctcggag gaggcctccc
     6601 agcccatggt cttcttctgc attacggggc cgtcggaggg gaagttggtg ccgcgcagct
     6661 tcaccttgta gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc
     6721 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttca
     6781 agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac atgaactgag
     6841 gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttggcgg
     6901 tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
     6961 tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat
     7021 cctcctcgcc cttgctcact ccggagccag ggccagggtt ctcctcgacg tcgccagcct
     7081 gcttcagcag ggaaaagttg gtggccccgg atccatcgaa ttcacgcgtc ttttgatgaa
     7141 acagaagttt tttgatattt ccatttgaat attttggtat ctgattggtg atgatttcag
     7201 ctaacatctc ggggaattca atactcatgg tcttatccaa aaatgtttgg aagcaatagt
     7261 taaggagatt ttcaaccact tcatgcatag aatccaagag ttttgtcagt tgataaaacc
     7321 gctgccagtt ctggctggag tttccttccc tcttgacaat ggcttttcct agctctttga
     7381 tgtaggtcat tctaatttca tcaaatagct cttggctctt cagaccgtcc ttaggaactg
     7441 aagagagaag cagtaaggtt ttcatacaga gatactcttc ataagatacc tgaagcctgt
     7501 gtaactcaga ggaaacatac agcatgtgtt tacattggtc gtacatgcag ggtagagtca
     7561 ttctctgctc attaataatc agatcaggag caaaacacag caggtttgca cttgattgtc
     7621 tatatgatct ccaccccaga gcaaatgcca taagaaacat ccaggagtac tgcagtaggg
     7681 tcatttggtc atccaggtgt aagttcctga aacctggtat tgcctttgcc catttcactg
     7741 ctgcaatcac ttgccgccct cctaacatgt tgagcgtagt catgatcctc caagttgagt
     7801 ctggaacaga gctatcatat cctgcatata acacttcagg ttcaataacc tccaacagtg
     7861 acaccagggt aggggtgagt tgtggtaacg ttgcaggaac tattgttttg ttaccaggat
     7921 tttcagaggt ttcttgtgag actcctgtag tggcctgctg aattcctttt atttttttct
     7981 ttgtttttcg agcttccagg ttcattccag cctgaagaca ttttcgatag cggcatgctg
     8041 ggcagttttt tcttcgaatt ttatcgatga tgcaatcatt ccttccagca cataggtaat
     8101 tgtgctgtcc ttccactgct cttttgaaga aaactttaca gcttccacaa gttaagactc
     8161 cataatgaca tcctgaagct tcatcagagc acaccaggca gagtttggga ggtggtcctg
     8221 ttgttgctgt tgaggagctg gatggaggag agcttacatc tggtctcatg ctggggcttg
     8281 aatagccatt agaaaaaact gttcgaccag ggaagttcag agtccccaga gaagtcaagt
     8341 tgtcatctcc agatccttgg cacctattcc aattttcgga accaacggga attggtggaa
     8401 tgacattaaa aataggcttc tgatcctgct gttgagaaag ggatgctgta ttcatgtcat
     8461 agtggtacat ctgtcctcca gaggtactca caccatgaac agaaatggca gacattttat
     8521 taccaattat atttgctcca ggaaagcttg cctgacagta aactgtgccc agtttctctt
     8581 gcttaattac cccaggggtg cagagttcga tgaaatcttc tttttctgtt ttcacttggg
     8641 gcagtgttac attactgggg cttgacaaaa ccagatctcc attatcctta attttgggtt
     8701 tagtgtccgg taaaatgaga ggcttgcagt cctcattcga gtttccttcc aaaaggaatg
     8761 aatcgtcttc tcccgccaga ggagaaagca aacagttttc atctatcaac aggtctgatc
     8821 tccaaggact ctcattcgtc tctttacctg gggacccaga agaaaactcc aaatcctgca
     8881 aaatgtcaaa ggtgctttgg tctgtggtat acaatttcac attgccaccg ttggtgccag
     8941 tctggccctt caaatgttgc tgttctgaag atacatcaga gtgagttttt ggaaactcct
     9001 tctctgtggg ggcagcagac acagcagtgg atgctgaact cttggggttc tctggaacac
     9061 tggtcgacct attgaggttt gcaatgcttt cttccaaaag ctttaagtct gtttcccccg
     9121 aggaaaggct gatttggccc tgctgtggga atcccaggtc atttcccatc acttttgttt
     9181 ctgtctctcc catatacagt cccattgaga gtgaaactgc tttggacaga tctggctgct
     9241 gcgcattgct tactgagcct tttggaaaat caaccaaaag tcttcgctgc ttggagtctg
     9301 attgagaagc gacagccagt gagggtgaag acgcagaaac cttcacagta gctcctcctc
     9361 ttagggtttt atagaagtcc atcacatctc ccctctcctg agcaagcaca ctgctggggt
     9421 tttcttctct accaggagtt aatgattctt tggagtccat ggtggcggaa ttcggggccg
     9481 cggaggctgg atcggtcccg gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc
     9541 caggcgatct gacggttcac taaacgagct ctgcttatat aggcctccca ccgtacacgc
     9601 ctacctcgac ccgggtaccg agctcgactt ccacttttct ctatcactga tagggagtgg
     9661 taaactcgac tttcactttt ctctatcact gatagggagt ggtaaactcg actttcactt
     9721 ttctctatca ctgataggga gtggtaaact cgactttcac ttttctctat cactgatagg
     9781 gagtggtaaa ctcgactttc nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     9841 nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     9901 acttttctct atcactgata gggagtggta aactcgagat ctaaccaaac tggatctctg
     9961 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
    10021 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
    10081 cccccttttc ttttaaaatt gtggatgaat actgccattt gtctgcagaa ttggcgcacg
    10141 cagtgccgat ccgttcacta atcgaatgga tctgtctctg tctctctctc caccttcttc
    10201 ttctattcct tcgggcctgt cgggtcccct cggggttggg aggtgggtct gaaacgataa
    10261 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
    10321 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
    10381 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
    10441 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
    10501 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
    10561 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
    10621 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
    10681 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
    10741 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
    10801 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
    10861 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
    10921 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
    10981 atctcctcct ccaggtctga agatcagcgg ccgcttgctg tgcggtggtc ttacttttgt
    11041 tttgctcttc ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat
    11101 gcacacaata gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct
    11161 gaagggatgg ttgtagctgt cccagtattt gtctacagcc ttctgatgtt tctaacaggc
    11221 caggattaac tgcgaatcgt tctagctccc tgcttgccca tactatatgt tttaatttat
    11281 attttttctt tccccctggc cttaaccgaa ttttttccca tcgcgatcta attctccccc
    11341 gcttaatact gacgctctcg cacccatctc tctccttcta gcctccgcta gtcaaaattt
    11401 ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag
    11461 tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc
    11521 actgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
    11581 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
    11641 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
    11701 cccagtacag
//
