LOCUS       sequence-384267-       10655 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-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 10655)
  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 10655)
  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-3G
FEATURES             Location/Qualifiers
     source          1..10655
                     /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        4772..4790
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4825..5390
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4840..5392
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4984..5001)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5211..5230
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5398..5400,5401..5403,5404..6114)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5398..5400,5401..5403,5404..6114)
                     /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(5443..5464)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5704..5723)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6051..6072
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6118..6492
                     /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    6499..7087
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6552..6572)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6970..6981)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7174..7407
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7479..7600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7516..7535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7570..7589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7640..7775
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(7796..7814)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7824..7841)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7824..7840)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7833..7855)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7982..8437
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8069..8088)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8279..8300
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8463..8567
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8568..8636,8637..9428)
                     /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(8786..8805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9599..10187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10088..10107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10341..10358
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10475..10496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10511..10516,10517..10534,10535..10541)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10549..10565
                     /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     10554..10576
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10573..10589
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10573..10589
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10608..10628
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10610..10628
                     /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 gatttttggg tgctggtggt ggttggggga gtcctggctt gctatagctt gctagtaaca
     4141 gtggccttta ttattttctg ggtgaggagt aagaggagca ggctcctgca cagtgactac
     4201 atgaacatga ctccccgccg ccccgggccc acccgcaagc attaccagcc ctatgcccca
     4261 ccacgcgact tcgcagccta tcgctccaga aaacggggca gaaagaaact cctgtatata
     4321 ttcaaacaac catttatgag accagtacaa actactcaag aggaagatgg ctgtagctgc
     4381 cgatttccag aagaagaaga aggaggatgt gaactgagag tgaagttcag caggagcgca
     4441 gacgcccccg cgtaccagca gggccagaac cagctctata acgagctcaa tctaggacga
     4501 agagaggagt acgatgtttt ggacaagaga cgtggccggg accctgagat ggggggaaag
     4561 ccgagaagga agaaccctca ggaaggcctg tacaatgaac tgcagaaaga taagatggcg
     4621 gaggcctaca gtgagattgg gatgaaaggc gagcgccgga ggggcaaggg gcacgatggc
     4681 ctttaccagg gtctcagtac agccaccaag gacacctacg acgcccttca catgcaggcc
     4741 ctgccccctc gctaacgtac gcccgcgaaa ttaatacgac tcactatagg gagaccacaa
     4801 cggtttccct ctagcgggat caattccgcc ccccccctaa cgttactggc cgaagccgct
     4861 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     4921 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     4981 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     5041 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     5101 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     5161 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     5221 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     5281 atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     5341 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tactagtatg
     5401 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     5461 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     5521 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     5581 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     5641 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     5701 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     5761 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     5821 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     5881 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     5941 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     6001 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     6061 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcatg
     6121 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     6181 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     6241 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     6301 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     6361 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     6421 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     6481 gagcaggact gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     6541 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6601 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6661 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6721 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6781 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6841 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     6901 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6961 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     7021 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     7081 tccccgcctg taattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag
     7141 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     7201 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     7261 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     7321 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     7381 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     7441 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     7501 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     7561 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     7621 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     7681 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7741 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     7801 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     7861 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     7921 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     7981 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     8041 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     8101 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     8161 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     8221 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     8281 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     8341 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     8401 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     8461 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     8521 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     8581 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     8641 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     8701 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     8761 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     8821 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     8881 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     8941 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     9001 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     9061 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     9121 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     9181 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     9241 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     9301 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     9361 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     9421 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     9481 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     9541 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     9601 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     9661 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     9721 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     9781 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     9841 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     9901 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     9961 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
    10021 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
    10081 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
    10141 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
    10201 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
    10261 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
    10321 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
    10381 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
    10441 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
    10501 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
    10561 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
    10621 actaaaggga acaaaagctg gagctgcaag cttaa
//
