LOCUS       sequence-403583-       11799 bp    DNA     circular SYN 14-APR-2025
DEFINITION  2nd generation lentiviral transfer plasmid for engineering cells to 
            express human CD19.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11799)
  AUTHORS   Hamilton JR, Chen E, Perez BS, Sandoval Espinoza CR, Kang MH, 
            Trinidad M, Ngo W, Doudna JA
  TITLE     In vivo human T cell engineering with enveloped delivery vehicles.
  JOURNAL   Nat Biotechnol. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z.
  PUBMED    38212493
REFERENCE   2  (bases 1 to 11799)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z."
COMMENT     Sequence Label: pJRH-1363 LV EF1a-CD19 IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..11799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(143..162)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        328..707
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        709..907
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     862..882
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             925..1105
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1152..1277
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1770..2003
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2188..2232
                     /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    2530..2647
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2757..3934
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2988..3925
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3882..3902
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    5666..6239
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5689..6241
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5833..5850)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6060..6079
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6253..6255,6256..6258,6259..6972)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6253..6255,6256..6258,6259..6969)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6298..6319)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6559..6578)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6906..6927
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6987..7575
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7040..7060)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7458..7469)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7647..7880
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7906..7923)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7912..8136
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8182..8610
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8269..8288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8479..8500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8619..8639)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8624..8953
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8804..8939
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8866..8885
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9001..9048
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9067..9441
                     /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"
     polyA_signal    9571..9692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9608..9627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9662..9681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9741..9757)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9741..9757)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9754..9776)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9765..9781
                     /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)."
     promoter        complement(join(9789..9795,9796..9813,9814..9819))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9834..9855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9972..9989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10143..10731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10223..10242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10902..11693,11694..11762))
                     /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     11525..11544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(11763..11799,1..68))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
       61 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
      121 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
      181 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
      241 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
      301 tgcttcgcga tgtacgggcc agatattgac attgattatt gactagttat taatagtaat
      361 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      421 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      481 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      541 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      601 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      661 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      721 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      781 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      841 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      901 taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag
      961 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1021 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1081 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1141 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1201 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1261 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1321 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1381 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1441 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1501 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1561 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1621 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     1681 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     1741 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     1801 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     1861 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     1921 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     1981 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2041 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2101 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2161 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2221 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2281 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2341 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2401 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     2461 attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt
     2521 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     2581 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     2641 aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta aggtaccatc
     2701 gatcttaagt cgcgactcga gaattcgcta gctaggtctt gaaaggagtg ggaattggct
     2761 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2821 gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2881 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2941 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     3001 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     3061 ccactggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
     3121 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
     3181 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
     3241 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
     3301 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
     3361 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
     3421 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
     3481 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     3541 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     3601 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     3661 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     3721 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     3781 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     3841 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     3901 tcaaagtttt tttcttccat ttcaggtgtc gtgaaccggt tctagagcgc tgccaccatg
     3961 ccacctcctc gcctcctctt cttcctcctc ttcctcaccc ccatggaagt caggcccgag
     4021 gaacctctag tggtgaaggt ggaagaggga gataacgctg tgctgcagtg cctcaagggg
     4081 acctcagatg gccccactca gcagctgacc tggtctcggg agtccccgct taaacccttc
     4141 ttaaaactca gcctggggct gccaggcctg ggaatccaca tgaggcccct ggccatctgg
     4201 cttttcatct tcaacgtctc tcaacagatg gggggcttct acctgtgcca gccggggccc
     4261 ccctctgaga aggcctggca gcctggctgg acagtcaatg tggagggcag cggggagctg
     4321 ttccggtgga atgtttcgga cctaggtggc ctgggctgtg gcctgaagaa caggtcctca
     4381 gagggcccca gctccccttc cgggaagctc atgagcccca agctgtatgt gtgggccaaa
     4441 gaccgccctg agatctggga gggagagcct ccgtgtgtcc caccgaggga cagcctgaac
     4501 cagagcctca gccaggacct caccatggcc cctggctcca cactctggct gtcctgtggg
     4561 gtaccccctg actctgcgtc caggggcccc ctctcctgga cccatgtgca ccccaagggg
     4621 cctaagtcat tgctgagcct agagctgaag gacgatcgcc cggccagaga tatgtgggta
     4681 atggagacgg gtctgttgtt gccccgggcc acagctcaag acgctggaaa gtattattgt
     4741 caccgtggca acctgaccat gtcattccac ctggagatca ctgctcggcc agtactatgg
     4801 cactggctgc tgaggactgg tggctggaag gtctcagctg tgactttggc ttatctgatc
     4861 ttctgcctgt gttcccttgt gggcattctt catcttcaaa gagccctggt cctgaggagg
     4921 aaaagaaagc gaatgactga ccccaccagg agattcttca aagtgacgcc tcccccagga
     4981 agcgggcccc agaaccagta cgggaacgtg ctgtctctcc ccacacccac ctcaggcctc
     5041 ggacgcgccc agcgttgggc cgcaggcctg gggggcactg ccccgtctta tggaaacccg
     5101 agcagcgacg tccaggcgga tggagccttg gggtcccgga gcccgccggg agtgggccca
     5161 gaagaagagg aaggggaggg ctatgaggaa cctgacagtg aggaggactc cgagttctat
     5221 gagaacgact ccaaccttgg gcaggaccag ctctcccagg atggcagcgg ctacgagaac
     5281 cctgaggatg agcccctggg tcctgaggat gaagactcct tctccaacgc tgagtcttat
     5341 gagaacgagg atgaagagct gacccagcca gtcgccagga caatggactt cctgagccct
     5401 catgggtcag cctgggaccc cagccgggaa gcaacctccc tggggtccca gtcctatgag
     5461 gatatgagag gaatcctgta tgcagccccc cagctccgct ccattcgggg ccagcctgga
     5521 cccaatcatg aggaagatgc agactcttat gagaacatgg ataatcccga tgggccagac
     5581 ccagcctggg gaggaggggg ccgcatgggc acctggagca ccaggtgaac gcgttaagtc
     5641 gacaaatgac aagtaagcgg ccgcgcccct ctccctcccc cccccctaac gttactggcc
     5701 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     5761 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     5821 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     5881 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     5941 accccccacc tggcgacagg tgcctctgcg gcccaaagcc acgtgtataa gatacacctg
     6001 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     6061 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     6121 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     6181 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     6241 atggccacac atatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     6301 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     6361 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     6421 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     6481 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     6541 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     6601 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     6661 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     6721 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     6781 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     6841 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     6901 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     6961 ctgtacaagt agcgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     7021 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     7081 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     7141 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     7201 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     7261 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     7321 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     7381 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     7441 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     7501 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     7561 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag
     7621 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     7681 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     7741 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7801 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7861 agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     7921 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     7981 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     8041 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     8101 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     8161 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     8221 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     8281 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     8341 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     8401 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     8461 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     8521 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     8581 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     8641 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     8701 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     8761 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     8821 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     8881 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     8941 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     9001 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     9061 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     9121 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     9181 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     9241 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     9301 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
     9361 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
     9421 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
     9481 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     9541 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     9601 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9661 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     9721 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     9781 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     9841 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     9901 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     9961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
    10021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
    10081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
    10141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
    10201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
    10261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
    10321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
    10381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
    10441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
    10501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
    10561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
    10621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
    10681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
    10741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
    10801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
    10861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
    10921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
    10981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
    11041 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
    11101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
    11161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
    11221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
    11281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
    11341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
    11401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
    11461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
    11521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
    11581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
    11641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
    11701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
    11761 atactcttcc tttttcaata ttattgaagc atttatcag
//
