LOCUS       sequence-384270-       10520 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.F1 in VL-VH order 
            and 218 linker (scFv). GFP-Zeo for selection and monitoring..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10520)
  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 10520)
  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-F1.218.CAR-2G
FEATURES             Location/Qualifiers
     source          1..10520
                     /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        4637..4655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4690..5255
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4705..5257
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4849..4866)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5076..5095
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5263..5265,5266..5268,5269..5979)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5263..5265,5266..5268,5269..5979)
                     /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(5308..5329)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5569..5588)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5916..5937
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5983..6357
                     /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    6364..6952
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6417..6437)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6835..6846)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7039..7272
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7344..7465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7381..7400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7435..7454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7505..7640
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7661..7679)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7689..7706)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7689..7705)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7698..7720)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7847..8302
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7934..7953)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8144..8165
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8328..8432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8433..8501,8502..9293)
                     /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(8651..8670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9464..10052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9953..9972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10206..10223
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10340..10361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10376..10381,10382..10399,10400..10406)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10414..10430
                     /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     10419..10441
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10438..10454
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10438..10454
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10473..10493
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10475..10493
                     /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 atcccagata tccagatgac ccagagccct
     3241 agctccctga gcgcctccgt gggcgacagg gtgaccatca catgcagagc ctctcagagc
     3301 atctctagct acctgaactg gtatcagcag aagcccggca aggcccctaa gctgctgatc
     3361 tacgcagcat ccgccctgca gtctggagtg ccatcccggt tctccggatc tggaagcggc
     3421 accgacttta ccctgacaat ctcctctctg cagccagagg atttcgccac atactattgt
     3481 cagcaggcct actatgaccc caccacattt ggccagggca caaaggtgga gatcaagggc
     3541 tccacctctg gatccggcaa gcccggatct ggcgagggat ccaccaaggg cgaggtgcag
     3601 ctgctggaga gcggaggagg actggtgcag ccaggaggca gcctgaggct gtcctgcgcc
     3661 gcctctggct tcacctttag ctcctacgca atgtcttggg tgcgccaggc acctggcaag
     3721 ggactggagt gggtgagcac catctccaca tctggcgact acaccacata tgccgattcc
     3781 gtgaagggcc ggttcaccat cagcagagac aactccaaga atacactgta tctgcagatg
     3841 aactctctgc gggccgagga caccgccgtg tactattgtg ccaagtccac agcctacttt
     3901 gattattggg gccagggcac cctggtgaca gtgtctagcg cggccgcaac cacgacgcca
     3961 gcgccgcgac caccaacacc ggcgcccacc atcgcgtcgc agcccctgtc cctgcgccca
     4021 gaggcgtgcc ggccagcggc ggggggcgca gtgcacacga gggggctgga cttcgcctgt
     4081 gatatctaca tctgggcgcc cttggccggg acttgtgggg tccttctcct gtcactggtt
     4141 atcacccttt actgcaaacg gggcagaaag aaactcctgt atatattcaa acaaccattt
     4201 atgagaccag tacaaactac tcaagaggaa gatggctgta gctgccgatt tccagaagaa
     4261 gaagaaggag gatgtgaact gagagtgaag ttcagcagga gcgcagacgc ccccgcgtac
     4321 cagcagggcc agaaccagct ctataacgag ctcaatctag gacgaagaga ggagtacgat
     4381 gttttggaca agagacgtgg ccgggaccct gagatggggg gaaagccgag aaggaagaac
     4441 cctcaggaag gcctgtacaa tgaactgcag aaagataaga tggcggaggc ctacagtgag
     4501 attgggatga aaggcgagcg ccggaggggc aaggggcacg atggccttta ccagggtctc
     4561 agtacagcca ccaaggacac ctacgacgcc cttcacatgc aggccctgcc ccctcgctaa
     4621 cgtacgcccg cgaaattaat acgactcact atagggagac cacaacggtt tccctctagc
     4681 gggatcaatt ccgccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     4741 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     4801 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     4861 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     4921 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     4981 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     5041 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     5101 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     5161 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     5221 gggacgtggt tttcctttga aaaacacgat gataatacta gtatggtgag caagggcgag
     5281 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     5341 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     5401 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     5461 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     5521 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     5581 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     5641 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     5701 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     5761 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     5821 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     5881 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     5941 gccgccggga tcactctcgg catggacgag ctgtacaagg gcatggccaa gttgaccagt
     6001 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     6061 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     6121 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     6181 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     6241 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     6301 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgagtc
     6361 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     6421 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     6481 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     6541 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     6601 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     6661 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     6721 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     6781 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     6841 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     6901 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctgtaatt
     6961 cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     7021 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     7081 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     7141 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     7201 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     7261 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     7321 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     7381 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     7441 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     7501 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     7561 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     7621 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
     7681 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     7741 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     7801 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     7861 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     7921 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     7981 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     8041 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     8101 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     8161 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     8221 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     8281 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     8341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     8401 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     8461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     8521 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     8581 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     8641 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     8701 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     8761 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     8821 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     8881 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     8941 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     9001 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     9061 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     9121 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     9181 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     9241 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     9301 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     9361 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     9421 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     9481 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     9541 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     9601 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     9661 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     9721 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     9781 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     9841 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     9901 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     9961 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
    10021 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
    10081 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
    10141 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
    10201 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
    10261 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
    10321 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
    10381 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
    10441 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
    10501 agctggagct gcaagcttaa
//
