LOCUS       sequence-384169-       10667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAR featuring: GMCSF (sig. peptide); CD8A & CD8A (hinge & TM); CD28,
            4-1BB & CD3-zeta (signaling domains); anti-CD19 from FMC63 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 10667)
  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 10667)
  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.CD19-FMC63.218.CAR-3G
FEATURES             Location/Qualifiers
     source          1..10667
                     /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        4784..4802
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4837..5402
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4852..5404
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4996..5013)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5223..5242
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5410..5412,5413..5415,5416..6126)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5410..5412,5413..5415,5416..6126)
                     /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(5455..5476)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5716..5735)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6063..6084
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6130..6504
                     /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    6511..7099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6564..6584)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6982..6993)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7186..7419
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7491..7612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7528..7547)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7582..7601
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7652..7787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7808..7826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7836..7853)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7836..7852)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7845..7867)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7994..8449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8081..8100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8291..8312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8475..8579
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8580..8648,8649..9440)
                     /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(8798..8817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9611..10199
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10100..10119
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10353..10370
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10487..10508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10523..10528,10529..10546,10547..10553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10561..10577
                     /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     10566..10588
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10585..10601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10585..10601
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10620..10640
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10622..10640
                     /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 atcccagaca tccagatgac acagactaca
     3241 tcctccctgt ctgcctctct gggagacaga gtcaccatca gttgcagggc aagtcaggac
     3301 attagtaaat atttaaattg gtatcagcag aaaccagatg gaactgttaa actcctgatc
     3361 taccatacat caagattaca ctcaggagtc ccatcaaggt tcagtggcag tgggtctgga
     3421 acagattatt ctctcaccat tagcaacctg gagcaagaag atattgccac ttacttttgc
     3481 caacagggta atacgcttcc gtacacgttc ggagggggga ctaagttgga aataacaggc
     3541 tccacctctg gatccggcaa gcccggatct ggcgagggat ccaccaaggg cgaggtgaaa
     3601 ctgcaggagt caggacctgg cctggtggcg ccctcacaga gcctgtccgt cacatgcact
     3661 gtctcagggg tctcattacc cgactatggt gtaagctgga ttcgccagcc tccacgaaag
     3721 ggtctggagt ggctgggagt aatatggggt agtgaaacca catactataa ttcagctctc
     3781 aaatccagac tgaccatcat caaggacaac tccaagagcc aagttttctt aaaaatgaac
     3841 agtctgcaaa ctgatgacac agccatttac tactgtgcca aacattatta ctacggtggt
     3901 agctatgcta tggactactg gggtcaagga acctcagtca ccgtctcctc agcggccgca
     3961 accacgacgc cagcgccgcg accaccaaca ccggcgccca ccatcgcgtc gcagcccctg
     4021 tccctgcgcc cagaggcgtg ccggccagcg gcggggggcg cagtgcacac gagggggctg
     4081 gacttcgcct gtgatttttg ggtgctggtg gtggttgggg gagtcctggc ttgctatagc
     4141 ttgctagtaa cagtggcctt tattattttc tgggtgagga gtaagaggag caggctcctg
     4201 cacagtgact acatgaacat gactccccgc cgccccgggc ccacccgcaa gcattaccag
     4261 ccctatgccc caccacgcga cttcgcagcc tatcgctcca gaaaacgggg cagaaagaaa
     4321 ctcctgtata tattcaaaca accatttatg agaccagtac aaactactca agaggaagat
     4381 ggctgtagct gccgatttcc agaagaagaa gaaggaggat gtgaactgag agtgaagttc
     4441 agcaggagcg cagacgcccc cgcgtaccag cagggccaga accagctcta taacgagctc
     4501 aatctaggac gaagagagga gtacgatgtt ttggacaaga gacgtggccg ggaccctgag
     4561 atggggggaa agccgagaag gaagaaccct caggaaggcc tgtacaatga actgcagaaa
     4621 gataagatgg cggaggccta cagtgagatt gggatgaaag gcgagcgccg gaggggcaag
     4681 gggcacgatg gcctttacca gggtctcagt acagccacca aggacaccta cgacgccctt
     4741 cacatgcagg ccctgccccc tcgctaacgt acgcccgcga aattaatacg actcactata
     4801 gggagaccac aacggtttcc ctctagcggg atcaattccg ccccccccct aacgttactg
     4861 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     4921 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     4981 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     5041 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     5101 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     5161 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     5221 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     5281 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     5341 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     5401 aatactagta tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     5461 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     5521 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     5581 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     5641 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     5701 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     5761 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     5821 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     5881 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     5941 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     6001 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     6061 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     6121 tacaagggca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
     6181 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     6241 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     6301 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     6361 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     6421 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     6481 ttcgtggccg aggagcagga ctgagtcgac aatcaacctc tggattacaa aatttgtgaa
     6541 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     6601 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     6661 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     6721 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     6781 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6841 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6901 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6961 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     7021 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     7081 ctttgggccg cctccccgcc tgtaattcga gctcggtacc tttaagacca atgacttaca
     7141 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     7201 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     7261 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     7321 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     7381 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     7441 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     7501 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     7561 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     7621 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     7681 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     7741 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
     7801 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     7861 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     7921 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     7981 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     8041 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     8101 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     8161 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     8221 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     8281 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     8341 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     8401 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     8461 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     8521 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     8581 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     8641 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     8701 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     8761 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     8821 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     8881 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     8941 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     9001 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     9061 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     9121 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     9181 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     9241 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     9301 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     9361 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     9421 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     9481 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     9541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     9601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     9661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     9721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     9781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     9841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     9901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     9961 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
    10021 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
    10081 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
    10141 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
    10201 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
    10261 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
    10321 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
    10381 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
    10441 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
    10501 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
    10561 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
    10621 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaa
//
