LOCUS       sequence-472748-       11470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of ComMAND base gene regulating EGFP-FMRP 
            (therapeutically relevant gene).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11470)
  AUTHORS   Love KS, Johnstone CP, Peterman EL, Gaglione S, Birnbaum ME, 
            Galloway KE
  TITLE     Model-guided design of microRNA-based gene circuits supports precise
            dosage of transgenic cargoes into diverse primary cells.
  JOURNAL   Cell Syst. 2025 Apr 22:101269. doi: 10.1016/j.cels.2025.101269.
  PUBMED    40300600
REFERENCE   2  (bases 1 to 11470)
  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: "Cell Syst. 
            2025 Apr 22:101269. doi: 10.1016/j.cels.2025.101269."
COMMENT     Sequence Label: 
            pLentiX1-[TRE3G-EGFP-FMRP-bGH]-EFS-rtTA-P2A-mRuby2-WPRE
FEATURES             Location/Qualifiers
     source          1..11470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    36..161
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    654..887
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1072..1116
                     /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    1383..1499
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    1526..1750
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(3601..3624)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(3667..4377,4378..4380,4381..4383))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3667..4377,4378..4380,4381..4383))
                     /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(3709..3730)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4058..4077
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4317..4338
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4380..4389
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4444..4822)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    4580..4598
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4616..4634
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4652..4670
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4688..4706
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4724..4742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4760..4778
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4796..4814
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(5003..5022)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        5132..5343
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5356..6099
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             6109..6165
                     /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             6166..6876
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     misc_feature    6890..7478
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6943..6963)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7361..7372)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7570..7803
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7875..7996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7912..7931)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7966..7985
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8036..8171
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8191..8210)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8192..8210)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8220..8237)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8220..8236)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8229..8251)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8378..8833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8465..8484)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8675..8696
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8859..8963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8964..9032,9033..9824)
                     /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(9182..9201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9995..10583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10484..10503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10737..10754
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10871..10892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10907..10912,10913..10930,10931..10937)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10945..10961
                     /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     10950..10972
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10969..10985
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10969..10985
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11004..11024
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11006..11024
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11052..11278
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11279..11459
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
       61 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      121 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
      181 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      241 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      301 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      361 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
      421 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
      481 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
      541 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
      601 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
      661 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
      721 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
      781 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
      841 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
      901 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
      961 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1021 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1081 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     1141 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     1201 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     1261 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     1321 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta
     1381 acttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1441 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaatt caaaatttta
     1501 tcgtccagtg tggtggctac tagcgccata gagcccaccg catccccagc atgcctgcta
     1561 ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag aatagaatga
     1621 cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac agtgggagtg
     1681 gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta
     1741 gaaggcacag gtgcaacgtt gttagggtac tccattcacc agcggttgca gcccatctac
     1801 gctgtctggc ttttccttct tctggttacg atctttaccc gtgcgcagcc ggctcccttc
     1861 actggaggca ctctgcaggg atccatcagc tgttcttcct ttagcctctc ttggattacg
     1921 tggacgatta tctgttcggg aatgatcgtc gtttcctttg aagcctcctc ctcttcctct
     1981 tcctccttgt cctcttcctc ctcctccacg ccgccgtccg tctcctctgc gcaggaagct
     2041 ctccctctct tcctctgttg gagctaatga ccaatcactg agttcgtctc tgtggtcaga
     2101 ttctgtttca gaagcatttg atgcttcaga attagttcct gaagtatatc caggaccgcg
     2161 tctgccgtgc cccctatttc tgtaaggtct actacctcga cccattcctt gaccatcatc
     2221 agtcacatag cctttttcct tatctgtacg atttggtggt ggtctagaac tagctccaat
     2281 ttgtcgcaac tgctcatcaa tttgtaatct ctccaaacgc aactggtcta cttcctttaa
     2341 atagttcagg tgataatcca aaagaacagt ggcattagcg atgctgtctt ttgttcccac
     2401 aaaaacaaat ggtaccatac cctgccaagc cttgggttca ggtttctggg gtccccctgc
     2461 tacaattgat gaaaccacta acaccctctg gacttttgta ctgttaggtt gagaaaaatg
     2521 ggtgttttcc tttgtatcta aatgtttctt ttcttcagag gagttaggtc caacccttga
     2581 attattggaa ggtagggaac ttggtggcat aatttcctct tcttgtggga cacttttctc
     2641 attttcagcc tcaatcctca ccctcacaac tcctgacttg tccacgatct cttgaatcag
     2701 ctttccattt tttcctatta ctttgcctac taagtttcgt ggaacctgta tgacatcttc
     2761 agcaaattcc agaaagcttc tagccttttt cactgcatct tgatcctctc cataaatatg
     2821 aaatgtgcag gtatcctcat ctaaatcaat agcagtgacg ccaggcactt ttctagcttg
     2881 ctgaatatta gcaccatgag taccaatagc taaacccatc agatcttctc gtacgataaa
     2941 ctgttcatga aatcttgagg caagctgcct tgaactctcc agttgtttac tggcttcttc
     3001 atttctcagt ataagagaca acttggtgcg cagacttcga aagtgcatgt caatcaacat
     3061 gtgggctcgc tttgaggtga cttcattgat ggacaaaatt accagctgat aattttctgg
     3121 atcataagtt acagagaagg caccaactgc ctttttaaaa tccttatgtg ctgattcttt
     3181 ggcacacatt tgtcgtaaat cttctggcac ctccagcttg atcttatgga aagtatcttt
     3241 tgtagcaggt ttgttgggat taacagatcg tagacgctca attgtgacaa tttcattata
     3301 cgtagcatca catgctgcat attctatcac ataaaactca ccctttatca tcctcacttt
     3361 agctaaccac caacagcaag gctctttttc atttgctctg gaataaacct caacttcatc
     3421 actttcgttt atatctttat tataacctac aggtggtggg aatctcacat catggaatgg
     3481 aatctgtctc tctggttgcc agttgttttc aaaagcaact gttatagaat cttcatggac
     3541 atcttttaca aatgccttgt agaaagcgcc attggagccc cgcacttcca ccaccagctc
     3601 cttgtcatcg tcgtccttgt agtcctccat cgaattcgaa gcttgagctc gagatctgag
     3661 tccggacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     3721 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag
     3781 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     3841 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     3901 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     3961 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     4021 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     4081 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     4141 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     4201 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     4261 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     4321 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     4381 catggtggcc attacagcgt ggatggcgtc tccaggcgat ctgacggttc actaaacgag
     4441 ctctttacga gggtaggaag tggtacggaa agttggtata agacaaaagt gttgtggaat
     4501 tgctccaggc gatctgacgg ttcactaaac gagctctgct tttataggcg cccaccgtac
     4561 acgcctaaag cttatacgtt ctctatcact gatagggagt aaactggata tacgttctct
     4621 atcactgata gggagtaaac tgtagatacg ttctctatca ctgataggga gtaaactggt
     4681 catacgttct ctatcactga tagggagtaa actccttata cgttctctat cactgatagg
     4741 gagtaaagtc tgcatacgtt ctctatcact gatagggagt aaactcttca tacgttctct
     4801 atcactgata gggagtaaac tcgaggaata gtacgaacat ggtagctagt ggatccttcg
     4861 tcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc agaacatcag
     4921 gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt cttccccgat
     4981 aacggagacc ggcacactgg cgcatatcgg tggtcatcat gcgccagctt tcatccccga
     5041 tatgcaccac cgggtaaagt tcacgcatat gaagctggac atcacctccc acaacctcga
     5101 ggtggaacag tacgctgata tctcttatac tgggcagagc gcacatcgcc cacagtcccc
     5161 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     5221 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     5281 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     5341 cagtaatgtg ccaccatgag ccggctggac aagagcaaag tgatcaacag cgccctggaa
     5401 ctgctgaacg gcgtgggcat cgagggcctg accacccgga agctggccca gaaactgggc
     5461 gtggaacagc ccaccctgta ctggcacgtg aagaacaagc gggccctgct ggacgccctg
     5521 cccatcgaga tgctggaccg gcaccacacc cacagctgcc ctctggaagg cgagagctgg
     5581 caggacttcc tgcggaacaa cgccaagagc tacagatgcg ccctgctgag ccaccgggac
     5641 ggcgccaaag tgcacctggg caccagaccc accgagaagc agtacgagac actggaaaac
     5701 cagctggcct tcctgtgcca gcagggcttc agcctggaaa acgccctgta cgccctgagc
     5761 gccgtgggcc acttcaccct gggctgcgtg ctggaagaac aggaacacca ggtcgccaaa
     5821 gaggaaagag agacacccac caccgacagc atgccccccc tgctgaagca ggccatcgag
     5881 ctgttcgaca gacagggcgc cgagcccgcc ttcctgttcg gcctggaact gatcatctgc
     5941 ggcctcgaga agcagctgaa gtgcgagagc ggcggaccca ccgacgccct ggacgacttc
     6001 gacctggaca tgctgcccgc cgatgccctg gatgattttg atctggatat gctccctgct
     6061 gacgctctcg atgacttcga tctcgatatg ctgccaggcg gaagcggagc tactaacttc
     6121 agcctgctga agcaggctgg agacgtggag gagaaccctg gacctatggt gtctaagggc
     6181 gaagagctga tcaaggaaaa tatgcgtatg aaggtggtca tggaaggttc ggtcaacggc
     6241 caccaattca aatgcacagg tgaaggagaa ggcaatccgt acatgggaac tcaaaccatg
     6301 aggatcaaag tcatcgaggg aggacccctg ccatttgcct ttgacattct tgccacgtcg
     6361 ttcatgtatg gcagccgtac ttttatcaag tacccgaaag gcattcctga tttctttaaa
     6421 cagtcctttc ctgagggttt tacttgggaa agagttacga gatacgaaga tggtggagtc
     6481 gtcaccgtca tgcaggacac cagccttgag gatggctgtc tcgtttacca cgtccaagtc
     6541 agaggggtaa actttccctc caatggtccc gtgatgcaga agaagaccaa gggttgggag
     6601 cctaatacag agatgatgta tccagcagat ggtggtctga ggggatacac tcatatggca
     6661 ctgaaagttg atggtggtgg ccatctgtct tgctctttcg taacaactta caggtcaaaa
     6721 aagaccgtcg ggaacatcaa gatgcccggt atccatgccg ttgatcaccg cctggaaagg
     6781 ttagaggaaa gtgacaatga aatgttcgta gtacaacgcg aacacgcagt tgccaagttc
     6841 gccgggcttg gtggtgggat ggacgagctg tacaagtaac aacgccgata atcaacctct
     6901 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     6961 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     7021 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     7081 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     7141 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     7201 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     7261 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     7321 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     7381 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     7441 tcagacgagt cggatctccc tttgggccgc ctccccgccg ctcgctgata tagggcccgg
     7501 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     7561 aggggggact ggaagggcta gctcactccc aacgaagaca agatctgctt tttgcttgta
     7621 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     7681 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     7741 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7801 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     7861 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7921 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7981 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     8041 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     8101 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     8161 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     8221 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     8281 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     8341 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     8401 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     8461 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     8521 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     8581 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     8641 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     8701 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     8761 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     8821 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     8881 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     8941 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     9001 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     9061 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     9121 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     9181 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     9241 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     9301 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     9361 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     9421 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     9481 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     9541 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     9601 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     9661 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     9721 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     9781 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     9841 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     9901 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     9961 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
    10021 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
    10081 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
    10141 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
    10201 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
    10261 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
    10321 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
    10381 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
    10441 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
    10501 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
    10561 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
    10621 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
    10681 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
    10741 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
    10801 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
    10861 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
    10921 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
    10981 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
    11041 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
    11101 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
    11161 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
    11221 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
    11281 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
    11341 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
    11401 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
    11461 tggcgcccga
//
