LOCUS       sequence-383064-        9912 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Triple reporter plasmid in pELPS with eDHFR (PET reporter gene) 
            fused to yellow fluorescent protein (YFP) and a T2A cleavage site 
            followed by Renilla luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9912)
  AUTHORS   Sellmyer MA, Richman SA, Lohith K, Hou C, Weng CC, Mach RH, O'Connor
            RS, Milone MC, Farwell MD
  TITLE     Imaging CAR T Cell Trafficking with eDHFR as a PET Reporter Gene.
  JOURNAL   Mol Ther. 2020 Jan 8;28(1):42-51. doi: 10.1016/j.ymthe.2019.10.007. 
            Epub 2019 Oct 15.
  PUBMED    31668558
REFERENCE   2  (bases 1 to 9912)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.ymthe.2019.10.007"; journalName: "Mol Ther"; date: 
            "2020-01-8- 8"; volume: "28"; issue: "1"; pages: "42-51"
COMMENT     Sequence Label: pELPS-eDHFR-YFP-T2A-Renilla
FEATURES             Location/Qualifiers
     source          1..9912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..1192
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          245..1183
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1140..1160
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(1688..1690,1691..2401)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1730..1751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1991..2010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2338..2359
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2411..2464
                     /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             2471..3403
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="TSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     misc_feature    3410..3998
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3463..3483)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3881..3892)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4085..4318
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4390..4511
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4427..4446)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4481..4500
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4551..4686
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4706..4725)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4707..4725)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4735..4752)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4735..4751)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4744..4766)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4893..5348
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4980..4999)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5190..5211
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5374..5478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5479..5547,5548..6339)
                     /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(5697..5716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6510..7098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6999..7018
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7252..7269
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7386..7407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7422..7427,7428..7445,7446..7452)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7460..7476
                     /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     7465..7487
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7484..7500
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7484..7500
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7519..7539
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7521..7539
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7567..7793
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7794..7974
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8021..8146
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8639..8872
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9057..9101
                     /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    9755..9872
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 attgatcacg tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
       61 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
      121 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
      181 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
      241 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
      301 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt
      361 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
      421 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
      481 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
      541 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
      601 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
      661 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
      721 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
      781 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
      841 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
      901 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc actgagtacc gggcgccgtc
      961 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     1021 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     1081 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     1141 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gatctagagg
     1201 atccatgata agtttgattg ctgctctggc tgtggaccgg gtaatcggta tggaaaacgc
     1261 catgccctgg aacctgcctg ccgatttggc ttggttcaag cgcaataccc tgaacaaacc
     1321 agtaatcatg ggaaggcata catgggaaag cattggaaga ccacttcccg gtagaaagaa
     1381 tattatcctg tctagccagc ccggcacgga tgatagggtg acatgggtaa agagcgtcga
     1441 tgaggcgatt gcggcgtgtg gtgacgtgcc ggaaattatg gttatcggag gcggcagggt
     1501 ctacgaacag ttcctgccga aggcacagaa gctgtacctc acccacatcg atgcagaggt
     1561 ggaaggagac acgcactttc cagattacga gcctgatgac tgggagagtg tttttagcga
     1621 attccatgac gcagacgccc aaaactctca ctcctactgc tttgagattc tcgaacgaag
     1681 ggcatgcgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1741 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1801 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1861 caccctcgtg accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat
     1921 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1981 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2041 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2101 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2161 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2221 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2281 ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2341 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2401 gggcagcgga gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     2461 ccctgctagc acttcgaaag tttatgatcc agaacaaagg aaacggatga taactggtcc
     2521 gcagtggtgg gccagatgta aacaaatgaa tgttcttgat tcatttatta attattatga
     2581 ttcagaaaaa catgcagaaa atgctgttat ttttttacat ggtaacgcgg cctcttctta
     2641 tttatggcga catgttgtgc cacatattga gccagtagcg cggtgtatta taccagacct
     2701 tattggtatg ggcaaatcag gcaaatctgg taatggttct tataggttac ttgatcatta
     2761 caaatatctt actgcatggt ttgaacttct taatttacca aagaagatca tttttgtcgg
     2821 ccatgattgg ggtgcttgtt tggcatttca ttatagctat gagcatcaag ataagatcaa
     2881 agcaatagtt cacgctgaaa gtgtagtaga tgtgattgaa tcatgggatg aatggcctga
     2941 tattgaagaa gatattgcgt tgatcaaatc tgaagaagga gaaaaaatgg ttttggagaa
     3001 taacttcttc gtggaaacca tgttgccatc aaaaatcatg agaaagttag aaccagaaga
     3061 atttgcagca tatcttgaac cattcaaaga gaaaggtgaa gttcgtcgtc caacattatc
     3121 atggcctcgt gaaatcccgt tagtaaaagg tggtaaacct gacgttgtac aaattgttag
     3181 gaattataat gcttatctac gtgcaagtga tgatttacca aaaatgttta ttgaatcgga
     3241 cccaggattc ttttccaatg ctattgttga aggtgccaag aagtttccta atactgaatt
     3301 tgtcaaagta aaaggtcttc atttttcgca agaagatgca cctgatgaaa tgggaaaata
     3361 tatcaaatcg ttcgttgagc gagttctcaa aaatgaacaa taagtcgaca atcaacctct
     3421 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3481 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3541 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3601 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3661 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3721 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3781 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3841 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3901 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3961 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacct
     4021 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     4081 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     4141 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     4201 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     4261 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     4321 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     4381 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4441 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4501 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc
     4561 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4621 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4681 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     4741 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4801 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4861 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4921 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4981 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5041 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5101 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5161 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5221 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5281 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5341 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5401 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5461 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5521 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5581 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5641 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5701 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5761 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5821 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5881 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5941 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6001 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6061 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6121 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6181 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6241 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6301 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6361 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6421 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6481 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6541 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6601 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6661 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6721 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6781 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6841 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6901 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6961 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7021 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7081 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7141 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7201 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7261 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7321 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7381 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7441 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     7501 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa
     7561 gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc
     7621 aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt
     7681 acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg
     7741 aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc
     7801 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7861 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7921 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     7981 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     8041 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     8101 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     8161 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     8221 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     8281 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     8341 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     8401 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     8461 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
     8521 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     8581 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     8641 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     8701 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     8761 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     8821 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     8881 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     8941 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     9001 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     9061 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     9121 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     9181 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     9241 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     9301 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc
     9361 gattagactg tagcccagga atatggcagc tagattgtac acatttagaa ggaaaagtta
     9421 tcttggtagc agttcatgta gccagtggat atatagaagc agaagtaatt ccagcagaga
     9481 cagggcaaga aacagcatac ttcctcttaa aattagcagg aagatggcca gtaaaaacag
     9541 tacatacaga caatggcagc aatttcacca gtactacagt taaggccgcc tgttggtggg
     9601 cggggatcaa gcaggaattt ggcattccct acaatcccca aagtcaagga gtaatagaat
     9661 ctatgaataa agaattaaag aaaattatag gacaggtaag agatcaggct gaacatctta
     9721 agacagcagt acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     9781 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     9841 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     9901 cagtttggct gc
//
