LOCUS       sequence-384263-       10511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAR featuring: GMCSF (sig. peptide); CD8A & CD8A (hinge & TM); 4-1BB
            & CD3-zeta (signaling domains); anti-JAG1 from J1.B5 in VH-VL order 
            and (GGGGS)3 linker (scFv). GFP-Zeo for selection and monitoring..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10511)
  AUTHORS   Silva G, Rodrigues AF, Ferreira S, Matos C, Eleuterio RP, Marques G,
            Kucheryava K, Lemos AR, Sousa PMF, Castro R, Barbas A, Simao D, 
            Alves PM
  TITLE     Novel scFv against Notch Ligand JAG1 Suitable for Development of 
            Cell Therapies toward JAG1-Positive Tumors.
  JOURNAL   Biomolecules. 2023 Mar 2;13(3):459. doi: 10.3390/biom13030459.
  PUBMED    36979394
REFERENCE   2  (bases 1 to 10511)
  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"; journalName: 
            "Biomolecules."; date: "2023-03-2"; volume: "13(3):459. doi"; pages:
            " 10.3390/biom13030459"
COMMENT     Sequence Label: pRRL.SIN.EF1A.JAG1-B5.GS.CAR-2G
FEATURES             Location/Qualifiers
     source          1..10511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1802..1919
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1944..3122
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2175..3113
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3070..3090
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        4628..4646
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4681..5246
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4696..5248
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4840..4857)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5067..5086
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5254..5256,5257..5259,5260..5970)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5254..5256,5257..5259,5260..5970)
                     /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(5299..5320)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5560..5579)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5907..5928
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5974..6348
                     /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"
     misc_feature    6355..6943
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6408..6428)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6826..6837)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7030..7263
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7335..7456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7372..7391)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7426..7445
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7496..7631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7652..7670)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7680..7697)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7680..7696)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7689..7711)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7838..8293
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7925..7944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8135..8156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8319..8423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8424..8492,8493..9284)
                     /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(8642..8661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9455..10043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9944..9963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10197..10214
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10331..10352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10367..10372,10373..10390,10391..10397)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10405..10421
                     /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     10410..10432
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10429..10445
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10429..10445
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10464..10484
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10466..10484
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg cctctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta
     1921 tcgatcacga gactagcctc gagggctccg gtgcccgtca gtgggcagag cgcacatcgc
     1981 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
     2041 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     2101 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     2161 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
     2221 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
     2281 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
     2341 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
     2401 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
     2461 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
     2521 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
     2581 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     2641 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
     2701 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     2761 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     2821 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     2881 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     2941 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     3001 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     3061 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     3121 gaggaattag cttggtacga attcgccacc atgcttctcc tggtgacaag ccttctgctc
     3181 tgtgagttac cacacccagc attcctcctg atcccagagg tgcagctgct ggagtccgga
     3241 ggaggactgg tgcagccagg aggatccctg aggctgtctt gcgccgccag cggcttcacc
     3301 tttagctcct acgcaatgtc ttgggtgcgc caggcacctg gcaagggact ggagtgggtg
     3361 agcacaatcg caggatccgg aaccgcaaca gactacgccg attctgtgaa gggccggttc
     3421 accatctcca gagacaactc taagaataca ctgtatctgc agatgaacag cctgcgggcc
     3481 gaggataccg ccgtgtacta ttgtgccaag gacacatacg cctttgatta ttggggccag
     3541 ggcaccctgg tgacagtgtc tagcggcggc ggaggcagcg gcggcggagg cagcggcggc
     3601 ggaggcagcg acatccagat gacccagagc cctagctccc tgagcgcctc cgtgggcgat
     3661 agggtgacca tcacatgcag agcctctcag agcatctcta gctacctgaa ctggtatcag
     3721 cagaagcccg gcaaggcccc taagctgctg atctacgcag caagctatct gcagtccgga
     3781 gtgccatctc ggttctccgg atctggaagc ggaaccgact ttaccctgac aatctcctct
     3841 ctgcagccag aggatttcgc cacatactat tgtcagcagg cctactccgc cccctctacc
     3901 tttggccagg gcacaaaggt ggagatcaag gcggccgcaa ccacgacgcc agcgccgcga
     3961 ccaccaacac cggcgcccac catcgcgtcg cagcccctgt ccctgcgccc agaggcgtgc
     4021 cggccagcgg cggggggcgc agtgcacacg agggggctgg acttcgcctg tgatatctac
     4081 atctgggcgc ccttggccgg gacttgtggg gtccttctcc tgtcactggt tatcaccctt
     4141 tactgcaaac ggggcagaaa gaaactcctg tatatattca aacaaccatt tatgagacca
     4201 gtacaaacta ctcaagagga agatggctgt agctgccgat ttccagaaga agaagaagga
     4261 ggatgtgaac tgagagtgaa gttcagcagg agcgcagacg cccccgcgta ccagcagggc
     4321 cagaaccagc tctataacga gctcaatcta ggacgaagag aggagtacga tgttttggac
     4381 aagagacgtg gccgggaccc tgagatgggg ggaaagccga gaaggaagaa ccctcaggaa
     4441 ggcctgtaca atgaactgca gaaagataag atggcggagg cctacagtga gattgggatg
     4501 aaaggcgagc gccggagggg caaggggcac gatggccttt accagggtct cagtacagcc
     4561 accaaggaca cctacgacgc ccttcacatg caggccctgc cccctcgcta acgtacgccc
     4621 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag cgggatcaat
     4681 tccgcccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     4741 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     4801 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     4861 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     4921 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     4981 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     5041 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     5101 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     5161 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     5221 ttttcctttg aaaaacacga tgataatact agtatggtga gcaagggcga ggagctgttc
     5281 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     5341 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     5401 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     5461 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     5521 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     5581 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     5641 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     5701 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     5761 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     5821 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     5881 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     5941 atcactctcg gcatggacga gctgtacaag ggcatggcca agttgaccag tgccgttccg
     6001 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     6061 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     6121 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     6181 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     6241 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     6301 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgagt cgacaatcaa
     6361 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     6421 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6481 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6541 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6601 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6661 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6721 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     6781 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6841 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6901 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgtaat tcgagctcgg
     6961 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     7021 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     7081 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     7141 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     7201 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7261 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     7321 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7381 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7441 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     7501 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     7561 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     7621 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     7681 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     7741 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     7801 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     7861 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     7921 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     7981 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     8041 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     8101 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     8161 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     8221 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     8281 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     8341 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     8401 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     8461 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     8521 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     8581 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     8641 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     8701 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     8761 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     8821 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     8881 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     8941 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     9001 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     9061 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     9121 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     9181 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     9241 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     9301 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     9361 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     9421 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     9481 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     9541 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     9601 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     9661 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     9721 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     9781 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     9841 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     9901 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     9961 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
    10021 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
    10081 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
    10141 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
    10201 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
    10261 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
    10321 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
    10381 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
    10441 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
    10501 tgcaagctta a
//
