LOCUS       sequence-384265-       10646 bp    DNA     circular SYN 15-APR-2025
DEFINITION  CAR featuring: GMCSF (sig. peptide); CD8A & CD8A (hinge & TM); CD28,
            4-1BB & CD3-zeta (signaling domains); anti-JAG1 from J1.F1 in VH-VL 
            order.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10646)
  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 10646)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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.GS.CAR-3G
FEATURES             Location/Qualifiers
     source          1..10646
                     /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        4763..4781
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4816..5381
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4831..5383
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4975..4992)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5202..5221
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5389..5391,5392..5394,5395..6105)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5389..5391,5392..5394,5395..6105)
                     /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(5434..5455)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5695..5714)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6042..6063
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6109..6483
                     /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    6490..7078
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6543..6563)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6961..6972)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7165..7398
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7470..7591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7507..7526)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7561..7580
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7631..7766
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7787..7805)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7815..7832)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7815..7831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7824..7846)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7973..8428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8060..8079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8270..8291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8454..8558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8559..8627,8628..9419)
                     /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(8777..8796)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9590..10178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10079..10098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10332..10349
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10466..10487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10502..10507,10508..10525,10526..10532)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10540..10556
                     /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     10545..10567
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10564..10580
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10564..10580
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10599..10619
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10601..10619
                     /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 ggagagcgga
     3241 ggaggactgg tgcagccagg aggcagcctg aggctgtcct gcgccgcctc tggcttcacc
     3301 tttagctcct acgcaatgtc ttgggtgcgc caggcacctg gcaagggact ggagtgggtg
     3361 agcaccatct ccacatctgg cgactacacc acatatgccg attccgtgaa gggccggttc
     3421 accatcagca gagacaactc caagaataca ctgtatctgc agatgaactc tctgcgggcc
     3481 gaggacaccg ccgtgtacta ttgtgccaag tccacagcct actttgatta ttggggccag
     3541 ggcaccctgg tgacagtgtc tagcggcggc ggaggcagcg gcggcggagg cagcggcggc
     3601 ggaggcagcg atatccagat gacccagagc cctagctccc tgagcgcctc cgtgggcgac
     3661 agggtgacca tcacatgcag agcctctcag agcatctcta gctacctgaa ctggtatcag
     3721 cagaagcccg gcaaggcccc taagctgctg atctacgcag catccgccct gcagtctgga
     3781 gtgccatccc ggttctccgg atctggaagc ggcaccgact ttaccctgac aatctcctct
     3841 ctgcagccag aggatttcgc cacatactat tgtcagcagg cctactatga ccccaccaca
     3901 tttggccagg gcacaaaggt ggagatcaag gcggccgcaa ccacgacgcc agcgccgcga
     3961 ccaccaacac cggcgcccac catcgcgtcg cagcccctgt ccctgcgccc agaggcgtgc
     4021 cggccagcgg cggggggcgc agtgcacacg agggggctgg acttcgcctg tgatttttgg
     4081 gtgctggtgg tggttggggg agtcctggct tgctatagct tgctagtaac agtggccttt
     4141 attattttct gggtgaggag taagaggagc aggctcctgc acagtgacta catgaacatg
     4201 actccccgcc gccccgggcc cacccgcaag cattaccagc cctatgcccc accacgcgac
     4261 ttcgcagcct atcgctccag aaaacggggc agaaagaaac tcctgtatat attcaaacaa
     4321 ccatttatga gaccagtaca aactactcaa gaggaagatg gctgtagctg ccgatttcca
     4381 gaagaagaag aaggaggatg tgaactgaga gtgaagttca gcaggagcgc agacgccccc
     4441 gcgtaccagc agggccagaa ccagctctat aacgagctca atctaggacg aagagaggag
     4501 tacgatgttt tggacaagag acgtggccgg gaccctgaga tggggggaaa gccgagaagg
     4561 aagaaccctc aggaaggcct gtacaatgaa ctgcagaaag ataagatggc ggaggcctac
     4621 agtgagattg ggatgaaagg cgagcgccgg aggggcaagg ggcacgatgg cctttaccag
     4681 ggtctcagta cagccaccaa ggacacctac gacgcccttc acatgcaggc cctgccccct
     4741 cgctaacgta cgcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
     4801 tctagcggga tcaattccgc ccccccccta acgttactgg ccgaagccgc ttggaataag
     4861 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     4921 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     4981 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5041 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5101 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5161 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5221 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     5281 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     5341 accacgggga cgtggttttc ctttgaaaaa cacgatgata atactagtat ggtgagcaag
     5401 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     5461 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     5521 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     5581 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     5641 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     5701 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     5761 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     5821 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     5881 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     5941 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     6001 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     6061 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcat ggccaagttg
     6121 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     6181 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     6241 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     6301 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     6361 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     6421 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     6481 tgagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     6541 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     6601 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     6661 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     6721 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     6781 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     6841 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     6901 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     6961 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     7021 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     7081 gtaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     7141 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     7201 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     7261 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     7321 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     7381 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     7441 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     7501 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     7561 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     7621 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     7681 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     7741 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     7801 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     7861 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     7921 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     7981 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     8041 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     8101 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     8161 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     8221 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     8281 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     8341 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     8401 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     8461 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     8521 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     8581 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     8641 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     8701 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     8761 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     8821 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     8881 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     8941 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     9001 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     9061 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     9121 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     9181 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     9241 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     9301 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     9361 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     9421 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     9481 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     9541 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     9601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     9661 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     9721 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     9781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     9841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     9901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     9961 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
    10021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
    10081 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
    10141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
    10201 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
    10261 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
    10321 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
    10381 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
    10441 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
    10501 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
    10561 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
    10621 aacaaaagct ggagctgcaa gcttaa
//
