LOCUS       sequence-402980-        9911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible lentiviral plasmid expressing EGFP-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 9911)
  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 9911)
  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-EGFP-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..9911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    49..67
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    91..109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    133..151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    175..193
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    217..235
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    292..409
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(707..751)
                     /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    936..1169
                     /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    1662..1787
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1834..2014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2032..2230)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        2232..2611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2783..2802
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2877..2981
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2982..3050,3051..3842)
                     /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(3200..3219)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4013..4601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4502..4521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4755..4772
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4889..4910
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4925..4930,4931..4948,4949..4955)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4963..4979
                     /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     4968..4990
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4987..5003
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4987..5003
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5052..5173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5063..5082)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5117..5136
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5303..5677)
                     /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(5696..5743)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5791..6120)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5805..5940
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5859..5878)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6105..6125
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6134..6562)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6244..6265)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6456..6475
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6608..6832
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6821..6838
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6864..7044
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     7595..7611
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7596..7612
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7615..8203
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7721..7732
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     8130..8150
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8226..8933)
                     /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(8504..8523)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8773..8791
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(8943..8999)
                     /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(join(9024..9734,9735..9737,9738..9740))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(9024..9734,9735..9737,9738..9740))
                     /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(9066..9087)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     9415..9434
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9674..9695
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      9737..9746
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(9826..9850)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(9847..9885)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 cacttttctc tatcactgat agggagtggt aaactcgact ttcacttttc tctatcactg
       61 atagggagtg gtaaactcga ctttcacttt tctctatcac tgatagggag tggtaaactc
      121 gactttcact tttctctatc actgataggg agtggtaaac tcgactttca cttttctcta
      181 tcactgatag ggagtggtaa actcgacttt cacttttctc tatcactgat agggagtggt
      241 aaactcgaga tctaaccaaa ctggatctct gctgtccctg taataaaccc gaaaattttg
      301 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
      361 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa
      421 tactgccatt tgtctgcaga attggcgcac gcagtgccga tccgttcact aatcgaatgg
      481 atctgtctct gtctctctct ccaccttctt cttctattcc ttcgggcctg tcgggtcccc
      541 tcggggttgg gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca
      601 ctatagaaag tacagcaaaa actattctta aacctaccaa gcctcctact atcattatga
      661 ataattttat ataccacagc caatttgtta tgttaaacca attccacaaa cttgcccatt
      721 tatctaattc caataattct tgttcattct tttcttgctg gttttgcgat tcttcaatta
      781 aggagtgtat taagcttgtg taattgttaa tttctctgtc ccactccatc caggtcgtgt
      841 gattccaaat ctgttccaga gatttattac tccaactagc attccaaggc acagcagtgg
      901 tgcaaatgag ttttccagag caaccccaaa tccccaggag ctgttgatcc tttaggtatc
      961 tttccacagc caggattctt gcctggagct gcttgatgcc ccagactgtg agttgcaaca
     1021 gatgctgttg cgcctcaata gccctcagca aattgttctg ctgctgcact ataccagaca
     1081 ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg
     1141 ctcccaagaa cccaaggaac aaagctccta ttcccactgc tcttttttct ctctgcacca
     1201 ctcttctctt tgccttggtg ggtgctactc ctaatggttc aatttttact actttatatt
     1261 tatataattc acttctccaa ttgtccctca tatctcctcc tccaggtctg aagatcagcg
     1321 gccgcttgct gtgcggtggt cttacttttg ttttgctctt cctctatctt gtctaaagct
     1381 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
     1441 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
     1501 tgtctacagc cttctgatgt ttctaacagg ccaggattaa ctgcgaatcg ttctagctcc
     1561 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
     1621 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
     1681 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
     1741 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
     1801 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
     1861 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     1921 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     1981 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaacgc gctgcttata
     2041 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga
     2101 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
     2161 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
     2221 aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     2281 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     2341 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     2401 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     2461 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     2521 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     2581 tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgcga
     2641 agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc
     2701 cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac acaagcaggg
     2761 agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg
     2821 ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg
     2881 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2941 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     3001 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     3061 cgctggtaaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     3121 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     3181 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3241 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3301 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3361 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3421 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3481 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3541 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3601 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3661 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3721 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3781 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3841 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3901 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3961 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4021 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4081 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4141 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4201 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4261 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4321 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4381 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4441 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4501 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4561 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4621 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4681 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4741 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4801 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4861 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4921 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4981 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgacggta
     5041 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     5101 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     5161 caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     5221 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     5281 aatcgaaatc tcgtagcacg tgtcagtcct gctcctcggc cacgaagtgc acgcagttgc
     5341 cggccgggtc gcgcagggcg aactcccgcc cccacggctg ctcgccgatc tcggtcatgg
     5401 ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc cgaccactcg gcgtacagct
     5461 cgtccaggcc gcgcacccac acccaggcca gggtgttgtc cggcaccacc tggtcctgga
     5521 ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg tcctccacga
     5581 agtcccggga gaacccgagc cggtcggtcc agaactcgac cgctccggcg acgtcgcgcg
     5641 cggtgagcac cggaacggca ctggtcaact tggccatggt ttagttcctc accttgtcgt
     5701 attatactat gccgatatac tatgccgatg attaattgtc aacacgtgct gatcagatcc
     5761 gaaaatggat atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc
     5821 tcctcactac ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa
     5881 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     5941 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     6001 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     6061 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     6121 attccacaga attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     6181 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     6241 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     6301 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     6361 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     6421 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     6481 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     6541 aatgcgccgc tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc
     6601 tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
     6661 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     6721 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     6781 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
     6841 gatcagcggg tttaaacggg ccctgctaga gattttccac actgactaaa agggtctgag
     6901 ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa gcactcaagg
     6961 caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct cccaggctca
     7021 gatctggtct aaccagagag acccagtaca gtccggatgc agctctcggg ccatgtgatg
     7081 aaatgctagg cggctgtcaa acctccactc taatacttct ctctccgggt catccatccc
     7141 atgcaggctc acagggtgta acaagcgggt gttctctcct tcattggctt cttctacctt
     7201 ctcttgctca actggtacta gttctgctag aacggtggag tacgggtaag aamskmmryt
     7261 ksctagcctt gtagcaccat ccaaaggtca gtggatatct gatccctggc cctggtgtgt
     7321 agttctgcca atcagggaag tagccttgtg tgtggtagat ccacagatca aggatatctt
     7381 gtcttcgttg ggagtgaatt agcccttcca gtcccccctt ttcttttaaa aagtggctaa
     7441 gatctacagc tgccttgtaa gtcattggtc ttaaaggtac ctgaggtgtg actggaaaac
     7501 ccacctcctc ctcctcttgt gcttctagcc aggcacaatc agcattggta gctgctgtat
     7561 tgctacttgt gattgctcca tgtttttcta ggtctcgagg tcgacggtat cgatgcgggg
     7621 aagcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc ggaagaggcc
     7681 gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc gaagggacgt
     7741 agcagaagga cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc caaggaaagg
     7801 acgatgattt ccccgacaac accacggaat tgtcagtgcc caacagccga gcccctgtcc
     7861 agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg gggaaagcga
     7921 aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg gttgcgtcag
     7981 caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc tcataaagag
     8041 acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     8101 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     8161 taccagtcaa tctttcacaa attttgtaat ccagaggttg attgtcgact taacgcgttc
     8221 tagattcact tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
     8281 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
     8341 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
     8401 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     8461 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     8521 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     8581 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     8641 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     8701 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     8761 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     8821 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     8881 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct cactccggag
     8941 ccagggccag ggttctcctc gacgtcgcca gcctgcttca gcagggaaaa gttggtggcc
     9001 ccggatccat cgaattcacg cgtcttgtac agctcgtcca tgccgagagt gatcccggcg
     9061 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
     9121 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     9181 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
     9241 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
     9301 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     9361 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     9421 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     9481 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg
     9541 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     9601 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     9661 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     9721 tcctcgccct tgctcaccat ggtggcggaa ttcggggccg cggaggctgg atcggtcccg
     9781 gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac
     9841 taaacgagct ctgcttatat aggcctccca ccgtacacgc ctacctcgac ccgggtaccg
     9901 agctcgactt c
//
