LOCUS       sequence-458375-       15545 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Single transcript cageIL-10 (mRens) + ER-renin (H2M) (Lenti).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15545)
  AUTHORS   Aldrete CA, Call CC, Sant'Anna LE, Vlahos AE, Pei J, Cong Q, Gao XJ
  TITLE     Orthogonalized human protease control of secreted signals.
  JOURNAL   Nat Chem Biol. 2025 Jan 15. doi: 10.1038/s41589-024-01831-x.
  PUBMED    39814991
REFERENCE   2  (bases 1 to 15545)
  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: "Nat Chem 
            Biol. 2025 Jan 15. doi: 10.1038/s41589-024-01831-x."
COMMENT     Sequence Label: CA-0230
            EF1a_Puro_ERret-H2M-Renin_T2A_mRens-ca...
FEATURES             Location/Qualifiers
     source          1..15545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(830..850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        835..1164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1015..1150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2298..2363
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2493..2513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2938..3072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3029..3048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3240..3873
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3920..4045
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4538..4771
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4956..5000
                     /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"
     promoter        5275..5486
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          5487..6425
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6382..6402
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             6449..7045
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6449..6468)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6945..6965
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             7046..7102
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8408..8428
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             8552..8605
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9890..9919
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             9920..9973
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9974..10684
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(10119..10137)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     10387..10406
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    10699..11287
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10752..10772)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11170..11181)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11492..12125
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(13604..13622)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(13605..13622)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(13912..13931)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(14145..14164)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     14264..14286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(14324..14342)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        14410..14514
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(14515..14583,14584..15375)
                     /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(14733..14752)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgtg gaatgtgtgt
      841 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      901 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1081 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1141 tttggaggcc taggcttttg caaaaagctt ggacacaaga caggcttgcg agatatgttt
     1201 gagaatacca ctttatcccg cgtcagggag aggcagtgcg taaaaagacg cggactcatg
     1261 tgaaatactg gtttttagtg cgccagatct ctataatctc gcgcaaccta ttttcccctc
     1321 gaacactttt taagccgtag ataaacaggc tgggacactt cacatgagcg aaaaatacat
     1381 cgtcacctgg gacatgttgc agatccatgc acgtaaactc gcaagccgac tgatgccttc
     1441 tgaacaatgg aaaggcatta ttgccgtaag ccgtggcggt ctgtaccggg tgcgttactg
     1501 gcgcgtgaac tgggtattcg tcatgtcgat accgtttgta tttccagcta cgatcacgac
     1561 aaccagcgcg agcttaaagt gctgaaacgc gcagaaggcg atggcgaagg cttcatcgtt
     1621 attgatgacc tggtggatac gggtggtact gcggttgcga ttcgtgaaat gtatccaaaa
     1681 gcgcactttg tcaccatctt cgcaaaaccg gctggtcgtc cgctggttga tgactatgtt
     1741 gttgatatcc cgcaagatac ctggattgaa cagccgtggg atatgggcgt cgtattcgtc
     1801 ccgccaatct ccggtcgcta atcttttcaa cgcctggcac tgccgggcgt tgttcttttt
     1861 aacttcaggc gggttacaat agtttccagt aagtattctg gaggctgcat ccatgacaca
     1921 ggcaaacctg agcgaaaccc tgttcaaacc ccgctttaaa catcctgaaa cctcgacgct
     1981 agtccgccgc tttaatcacg gcgcacaacc gcctgtgcag tcggcccttg atggtaaaac
     2041 catccctcac tggtatcgca tgattaaccg tctgatgtgg atctggcgcg gcattgaccc
     2101 acgcgaaatc ctcgacgtcc aggcacgtat tgtgatgagc gatgccgaac gtaccgacga
     2161 tgatttatac gatacggtga ttggctaccg tggcggcaac tggatttatg agtgggcccc
     2221 ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag
     2281 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga
     2341 ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt
     2401 ggaatgcctt taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg
     2461 aggctactgc tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc
     2521 ccaaggactt tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa
     2581 ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa
     2641 ttatggaaaa atattctgta acctttataa gtaggcataa cagttataat cataacatac
     2701 tgttttttct tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat
     2761 tgtgtacctt tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg
     2821 ccttgactag agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2881 aacctcccac acctccccct gaacctgaaa cataaaatga atgcatttgt tgttgttaac
     2941 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3001 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3061 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     3121 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
     3181 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt
     3241 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
     3301 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
     3361 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
     3421 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     3481 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
     3541 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
     3601 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
     3661 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
     3721 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     3781 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     3841 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     3901 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     3961 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     4021 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     4081 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     4141 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     4201 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     4261 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     4321 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     4381 agccgccgct gatcttcaga cctggaggag gagatatgag ggacatttgg agaagtgaat
     4441 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     4501 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     4561 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     4621 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     4681 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     4741 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     4801 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     4861 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     4921 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     4981 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     5041 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     5101 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     5161 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     5221 acagatccat tcgattagtg aacggatctc gacggtatcg atgtgcccgt cagtgggcag
     5281 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     5341 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     5401 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     5461 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     5521 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
     5581 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
     5641 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
     5701 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
     5761 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
     5821 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
     5881 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
     5941 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
     6001 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
     6061 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
     6121 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     6181 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgcgcttt tggagtacgt
     6241 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     6301 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     6361 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     6421 ttcaggtgtc gtgagctagc tcgccaccat gaccgagtac aagcccacgg tgcgcctcgc
     6481 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     6541 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     6601 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     6661 cgccgtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     6721 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     6781 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc
     6841 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     6901 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     6961 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     7021 gtgcatgacc cgcaagcccg gtgccgccac gaacttctct ctgttaaagc aagcaggtga
     7081 cgtggaagaa aaccccggtc ccatggacgg ttggagaagg atgccccggt ggggcctttt
     7141 gctgctcctg tggggaagct gcaccttcgg ggctagcctc actctcggta atactacttc
     7201 ctcagtcatc ttaaccaact acatggatac gcagtattac ggggaaatcg gcattggcac
     7261 accccctcag acttttaagg tagtttttga tactgggtct agcaatgttt gggtccctag
     7321 cagtaagtgt tcccgcctgt atacagcgtg tgtttaccat aagttatttg acgcctcaga
     7381 ctcctctagc tataaacaca acggcaccga gctcaccatc cactacagca cagggactgt
     7441 ttcggggttt ctgtcccagg acattattac agtgggaggt attacagtga cccaaatgtt
     7501 tggggaggtg accgagatgc cggctctgcc gttcatgctg gcagagttcg acggggtcgt
     7561 tggcatgggc ttcatcgagc aggctgtggg gcgggtaacc cccatcttcg acaacatcat
     7621 tagccagggc gtgctaaagg aggacgtgtt cagcttttat tacaatagag attctgaaaa
     7681 ctctcagagt ttgggcggcc agatagtgct tggaggcagt gatccccagc actacgaagg
     7741 caattttcac tatataaatc tgatcaaaac tggggtgtgg cagatccaga tgaagggggt
     7801 gtcagtgggt tcctctacgt tattgtgcga ggacggatgt ctggccgtgg tagacactgg
     7861 agctagttac atcagcggct caaccagctc aattgaaaag ctgatggagg cactgggagc
     7921 aaaaaagcgg ttgttcgatt acgtcgtgaa atgtaatgag ggacctaccc tgcccgacat
     7981 ctctttccat ctgggtggaa aagagtatac tctgacaagc gccgattacg tatttcagga
     8041 gtcctactcg tccaaaaaac tctgcacgtt agcaatccac gccatggaca tccctccacc
     8101 aaccggacct acctgggctc tgggggccac atttatacga aaattctata ccgagttcga
     8161 tcgcaggaac aaccgtatcg gctttgcact tgccaggggc gccgggacag gagggtctgg
     8221 gggtggcact ggcggatctg gaggagacta caaggacgat gacgacaacg gcggatcggg
     8281 ggggtctggt ggtagtggcg gttcgggggg aggcacaggt ggatccatgg ctctcattgt
     8341 cttgggcggt gtggccgggt tactgctgtt catcggactt ggcatatttt tcgggggcgg
     8401 ctctctcgag aacttgtact tccagagtgg cagtggaggt acccttctcg aagccctgac
     8461 cttagccagt gcacggggac ccctgaggaa gagatctgtg ccaatggcta aggcgaaacc
     8521 gaaattctcc atctcacctg acagcttgag cgaaggtcgg gggagtttgc ttacctgtgg
     8581 cgatgtcgag gagaatccag gacccatgca ctcctccgcg ctcttgtgtt gcctagtttt
     8641 attgaccgga gtccgcgcat cacctgggca aggcactcag agtgaaaatt cttgcactca
     8701 cttccctgga attctcccca acatgctcag agacctgcga gatgctttct ctagagtcaa
     8761 aacgttcttt caaatgaagg accagctaga taatcttttg ctgaaagagt cccttctcga
     8821 ggattttaaa ggctatctgg gctgtcaggc tcttagcgag atgatacagt tctacctcga
     8881 ggaggtgatg ccacaggctg agaaccaaga cccagacatt aaagcccatg taatctcact
     8941 gggggaaaac ctgaacacat tacgcctgcg gctgcggagg tgccataggc tgctgccttg
     9001 tgaaaatggt gggggcagtg gaggcaagag caaggccgtg gaacaggtga agaatgcctt
     9061 taataagctg caggaaaagg ggatctacaa ggcaatgagc gagtttgata tattcatcaa
     9121 ctatatcgag gcctacatga ccatgaaaat tcgtaacggc gcctacatcc atcccttcca
     9181 cctgctctac cacaacaaga gcggcggtgg gggatctggt ggagggggat caggaggtgg
     9241 cggatcccat ggcactgagc ttccatctcc tccttctgta tggttcgaag cagagttctt
     9301 ccatcacatt ctgcattgga ctcccattcc aaaccaatcg gagagcacct gctacgaagt
     9361 ggctctgtta agatacggca ttgagtcttg gaacagcatc agcaactgct cacaaacgct
     9421 ctcttatgat ttgaccgcgg tgacacttga cctctaccac tcaaacggct atagagcccg
     9481 ggttcgagcc gtggatggct cccgacactc caactggacc gtcactaata cacggtttag
     9541 cgtggacgag gtgaccctga cagtgggaag tgtgaacctc gaaattcata acgggtttat
     9601 tctgggaaaa atacagctgc ctcgtccaaa gatggcacca gccaacgaca cgtacgaaag
     9661 cattttcagc cacttccggg agtacgaaat cgcaatccgc aaggtaccgg gtaattttac
     9721 ttttacccac aagaaggtta agcacgaaaa cttcagcttg ttaacctctg gcgaggtcgg
     9781 agagttttgc gtgcaagtga agcctagcgt cgcttcacgc tccaataaag gtatgtggag
     9841 taaagaggag tgcatcagtc tcacaaggca gtattttacg gtcaccaatg agcagaagct
     9901 catcagcgaa gaggacctgg agggacgggg aagtttgctt acctgtggcg atgtcgagga
     9961 gaatccaggt cccatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
    10021 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
    10081 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
    10141 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
    10201 ctacgtgaag caccccgccg acattcccga ctacttgaag ctgtccttcc ccgagggctt
    10261 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
    10321 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
    10381 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
    10441 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
    10501 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
    10561 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
    10621 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
    10681 gtagtacgaa ttcccgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
    10741 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
    10801 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
    10861 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
    10921 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
    10981 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
    11041 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
    11101 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
    11161 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
    11221 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
    11281 tccccgcatc gggatttctg cagtcgagac ctagaaaaac atggagcaat cacaagtagc
    11341 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
    11401 ggttttccag tcacacctca ggttccttta agaccaatga cttacaaggc agctgtagat
    11461 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
    11521 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
    11581 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
    11641 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
    11701 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
    11761 agccgcctag catttcatca catggcccga gagctgcatc cggagtactt caagaactgc
    11821 tgacatcgag cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg
    11881 ggcgggactg gggagtggcg agccctcaga tgctgcatat aagcagctgc tttttgcttg
    11941 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
    12001 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
    12061 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
    12121 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
    12181 atcagagagt gagaggcctt gacattataa tagatttagc aggaattgaa ctaggagtgg
    12241 agcacacagg caaagctgca gaagtacttg gaagaagcca ccagagatac tcacgattct
    12301 gcacatacct ggctaatccc agatcctaag gattacatta agtttactaa catttatata
    12361 atgatttata gtttaaagta taaacttatc taatttacta ttctgacaga tattaattaa
    12421 tcctcaaata tcataagaga tgattactat tatccccatt taacacaaga ggaaactgag
    12481 agggaaagat gttgaagtaa ttttcccaca attacagcat ccgttagtta cgactctatg
    12541 atcttctgac acaaattcca tttactcctc accctatgac tcagtcgaat atatcaaagt
    12601 tatggacatt atgctaagta acaaattacc cttttatata gtaaatactg agtagattga
    12661 gagaagaaat tgtttgcaaa cctgaatagc ttcaagaaga agagaagtga ggataagaat
    12721 aacagttgtc atttaacaag ttttaacaag taacttggtt agaaagggat tcaaatgcat
    12781 aaagcaaggg ataaattttt ctggcaacaa gactatacaa tataacctta aatatgactt
    12841 caaataattg ttggaacttg ataaaactaa ttaaatatta ttgaagatta tcaatattat
    12901 aaatgtaatt tacttttaaa aagggaacat agaaatgtgt atcattagag tagaaaacaa
    12961 tccttattat cacaatttgt caaaacaagt ttgttattaa cacaagtaga atactgcatt
    13021 caattaagtt gactgcagat tttgtgtttt gttaaaatta gaaagagata acaacaattt
    13081 gaattattga aagtaacatg taaatagttc tacatacgtt cttttgacat cttgttcaat
    13141 cattgatcga agttctttat cttggaagaa tttgttccaa agactctgaa ataaggaaaa
    13201 caatctatta tatagtctca cacctttgtt ttacttttag tgatttcaat ttaataatgt
    13261 aaatggttaa aatttattct tctctgagat catttcacat tgcagataga aaacctgaga
    13321 ctggggtaat ttttattaaa atctaattta atctcagaaa cacatcttta ttctaacatc
    13381 aatttttcca gtttgatatt atcatataaa gtcagccttc ctcatctgca ggttccacaa
    13441 caaaaatcca accaactgtg gatcaaaaat attgggaaaa aattaaaaat agcaatacaa
    13501 caataaaaaa atacaaatca gaaaaacagc acagtataac aactttattt agcatttaca
    13561 atctattagg tattataagt aatctagaat taattccgtg tattctatag tgtcacctaa
    13621 atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct aacgacaata
    13681 tgtacaagcc taattgtgta gcatctggct tactgaagca gaccctatca tctctctcgt
    13741 aaactgccgt cagagtcggt ttggttggac gaaccttctg agtttctggt aacgccgtcc
    13801 cgcacccgga aatggtcagc gaaccaatca gcagggtcat cggtagccag atcctctacg
    13861 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
    13921 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
    13981 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
    14041 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
    14101 tgcaggagtc gcataaggga gagcgtcgaa tggtgcactc tcagtacaat ctgctctgat
    14161 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
    14221 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
    14281 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
    14341 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
    14401 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
    14461 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
    14521 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
    14581 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
    14641 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
    14701 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
    14761 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
    14821 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
    14881 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
    14941 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
    15001 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
    15061 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
    15121 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
    15181 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
    15241 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
    15301 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
    15361 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
    15421 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
    15481 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
    15541 tcttc
//
