LOCUS       sequence-424739-        9968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  16xGCN4pep transgene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9968)
  TITLE     Cheeseman lab plasmids Nov 2023
REFERENCE   2  (bases 1 to 9968)
  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"
COMMENT     Sequence Label: pGS382_tdTomato-16xGCN4-T2A-BSD Lenti
FEATURES             Location/Qualifiers
     source          1..9968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    100..121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(136..141,142..159,160..166)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    174..190
                     /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     179..201
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     198..214
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     198..214
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     233..253
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        235..253
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        281..507
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             508..688
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    735..860
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1357..1590
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1775..1819
                     /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"
     primer_bind     1997..2017
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    2218..2335
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2407..2618
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2645..2654
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2651..4078
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             4124..4144
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4157..4201
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4238..4303
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4319..4384
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4400..4465
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4481..4546
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4562..4627
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4643..4708
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4724..4789
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4805..4870
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4886..4951
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             4967..5032
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             5048..5113
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             5129..5194
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     primer_bind     5210..5232
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     primer_bind     5233..5255
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             5258..5323
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             5339..5404
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             5420..5485
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             5501..5566
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             5597..5650
                     /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             5657..6055
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    6101..6689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6154..6174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6572..6583)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6761..6994
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7072..7193
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7109..7128)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7163..7182
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7233..7368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7388..7407)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7389..7407)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7417..7434)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7417..7433)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7426..7448)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7575..8030
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7662..7681)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7872..7893
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8056..8160
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8161..8229,8230..9021)
                     /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(8379..8398)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9192..9780
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9681..9700
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9934..9951
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
       61 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      121 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      181 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
      241 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
      301 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
      361 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
      421 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
      481 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
      541 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
      601 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
      661 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
      721 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
      781 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
      841 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
      901 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
      961 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     1021 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     1081 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     1141 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     1201 ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     1261 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     1321 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     1381 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     1441 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     1501 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     1561 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     1621 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     1681 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     1741 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     1801 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     1861 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     1921 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     1981 cccgagggga cccagagagg gcctatttcc catgattcct tcatatttgc atatacgata
     2041 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
     2101 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     2161 taaaatggac tatcataaat tctagatctt gagacaaatg gcagtattca tccacaattt
     2221 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     2281 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     2341 ttattacagg gacagcagag atccactttg gcgccggctc gtcgagtggc tccggtgccc
     2401 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     2461 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     2521 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     2581 acgttctttt tcgcaacggg tttgccgcca gaacacaggt gtcgtgacgc gggatccgat
     2641 atccgccacc atggtgagca agggcgagga ggtcatcaaa gagttcatgc gcttcaaggt
     2701 gcgcatggag ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg
     2761 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt
     2821 cgcctgggac atcctgtccc cccagttcat gtacggctcc aaggcgtacg tgaagcaccc
     2881 cgccgacatc cccgattaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     2941 gatgaacttc gaggacggcg gtctggtgac cgtgacccag gactcctccc tgcaggacgg
     3001 cacgctgatc tacaaggtga agatgcgcgg caccaacttc ccccccgacg gccccgtaat
     3061 gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt
     3121 gctgaagggc gagatccacc aggccctgaa gctgaaggac ggcggccact acctggtgga
     3181 gttcaagacc atctacatgg ccaagaagcc cgtgcaactg cccggctact actacgtgga
     3241 caccaagctg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg
     3301 ctccgagggc cgccaccacc tgttcctggg gcatggcacc ggcagcaccg gcagcggcag
     3361 ctccggcacc gcctcctccg aggacaacaa catggccgtc atcaaagagt tcatgcgctt
     3421 caaggtgcgc atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
     3481 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct
     3541 gcccttcgcc tgggacatcc tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa
     3601 gcaccccgcc gacatccccg attacaagaa gctgtccttc cccgagggct tcaagtggga
     3661 gcgcgtgatg aacttcgagg acggcggtct ggtgaccgtg acccaggact cctccctgca
     3721 ggacggcacg ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc
     3781 cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga
     3841 cggcgtgctg aagggcgaga tccaccaggc cctgaagctg aaggacggcg gccactacct
     3901 ggtggagttc aagaccatct acatggccaa gaagcccgtg caactgcccg gctactacta
     3961 cgtggacacc aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     4021 cgagcgctcc gagggccgcc accacctgtt cctgtacggc atggacgagc tgtacaagac
     4081 cggaggggat cggtggagct ccaccggtgg cggccgctct agagagaatc tttattttca
     4141 gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc acatggacag
     4201 cggaggtgga ggtactagtc tcgagggttc tggaggattg ttgcccaaaa actaccactt
     4261 ggagaacgag gtcgcaaggt tgaaaaaatt ggtcggtgag cgtggcggct caggcggcct
     4321 tcttccaaaa aattatcatc ttgaaaatga agttgctaga cttaaaaaac ttgttggaga
     4381 aagaggcggg agcggcggcc tgctgcccaa gaactaccac ctggagaacg aggtcgcccg
     4441 cctgaagaag ctggtcggcg agcgcggtgg ttctggaggt ttgctcccaa agaactacca
     4501 cttggaaaac gaagtcgcaa gattgaagaa gttggtcggt gaaagaggag gaagcggagg
     4561 attattacct aaaaattatc atttggaaaa tgaagttgct agattgaaga agttggttgg
     4621 agagagaggg gggtctggag gactactacc gaaaaactac cacctagaga acgaggtggc
     4681 ccgactaaaa aagctagtgg gcgagcgcgg cggcagtggc ggcctcctcc ccaagaatta
     4741 ccacctcgaa aatgaagtag caaggctcaa gaagctcgta ggagaaagag gtggttccgg
     4801 gggactcctt ccaaaaaact atcatcttga aaacgaagtt gcacgtctta aaaaacttgt
     4861 tggcgaacgt ggcggctctg gcgggctgct gcccaagaat taccacctgg aaaatgaagt
     4921 ggcccgcctg aagaagctgg tgggcgaacg cggaggcagt ggaggcctac taccgaagaa
     4981 ctaccaccta gagaacgagg tggcacgcct aaagaagcta gtgggcgagc gcggcggctc
     5041 gggtggttta ttacctaaga attatcattt agaaaatgaa gtagctcgat taaagaagtt
     5101 agtaggtgaa cgtggtgggt ctggtggtct tcttccgaag aactaccacc ttgagaacga
     5161 ggtagctcgt cttaagaaac ttgtaggtga gcgtggtgga agcggtggtg ggctggcaag
     5221 ccacgtttgg tgccgggagc tgcatgtgtc agaggaactt ttgccaaaga attatcatct
     5281 tgagaacgaa gtggctcgtc ttaagaaact cgtaggagaa agaggtggca gtggaggtct
     5341 gcttcccaag aattaccacc tggaaaatga ggtagctaga ctgaaaaagc tcgtaggcga
     5401 gcgcggggga agtgggggat tgctgccaaa aaattatcat ttggagaacg aagtagcacg
     5461 actaaagaaa ttagttgggg agaggggggg atcgggtggc ttactcccga aaaattatca
     5521 tctcgaaaac gaagtggctc ggctaaaaaa gttagttgga gagcgtggcg gttctggagg
     5581 cctattacca gaattcgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa
     5641 tcccggccct atcgatatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     5701 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     5761 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     5821 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     5881 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     5941 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     6001 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaaggtac
     6061 cactagtccc gggatgcata gcgctacgcg ttaagtcgac aatcaacctc tggattacaa
     6121 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     6181 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     6241 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     6301 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     6361 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6421 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6481 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     6541 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6601 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6661 tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca
     6721 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     6781 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     6841 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6901 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     6961 agaccctttt agtcagtgtg gaaaatctct agcagtacgt atagtagttc atgtcatctt
     7021 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     7081 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     7141 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     7201 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     7261 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7321 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     7381 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     7441 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     7501 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     7561 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     7621 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     7681 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     7741 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     7801 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     7861 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     7921 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     7981 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     8041 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     8101 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     8161 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     8221 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     8281 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     8341 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     8401 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     8461 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     8521 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     8581 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     8641 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     8701 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     8761 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     8821 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     8881 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     8941 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     9001 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     9061 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     9121 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     9181 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     9241 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     9301 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     9361 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     9421 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     9481 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     9541 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     9601 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     9661 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     9721 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     9781 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     9841 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     9901 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     9961 gagcgccc
//
