LOCUS       sequence-472751-       10723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of ComMAND closed-loop circuit regulating 
            FXN-P2A-mRuby2 (therapeutically relevant gene).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10723)
  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 10723)
  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-FXN-P2A-mRuby2(miRE.FF4)-TS.FF4x1-bGH]-EFS-rtTA-P2A-
            mGL-WPRE
FEATURES             Location/Qualifiers
     source          1..10723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    9..233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     ncRNA           complement(996..1108)
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           complement(1165..1179)
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     CDS             complement(1418..1474)
                     /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"
     promoter        complement(2174..2552)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2310..2328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2346..2364
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2382..2400
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2418..2436
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2454..2472
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2490..2508
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2526..2544
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(2733..2752)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        2862..3073
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3086..3829
                     /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             3839..3895
                     /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             join(3896..3898,3899..3901,3902..4615)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     primer_bind     complement(3941..3962)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    4626..5214
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4679..4699)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5097..5108)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5306..5539
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5611..5732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5648..5667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5702..5721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5772..5907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5927..5946)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5928..5946)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5956..5973)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5956..5972)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5965..5987)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6114..6569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6201..6220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6411..6432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6595..6699
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6700..6768,6769..7560)
                     /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(6918..6937)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7731..8319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8220..8239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8473..8490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8607..8628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8643..8648,8649..8666,8667..8673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8681..8697
                     /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     8686..8708
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8705..8721
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8705..8721
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8740..8760
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8742..8760
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8788..9014
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9015..9195
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9242..9367
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9860..10093
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10278..10322
                     /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    10589..10705
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tactagcgcc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
       61 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
      121 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
      181 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca caggtgcaac
      241 tcctcctagg tttaattaaa gacttcaagc ggccacaagc ttgttgttac ttgtacagct
      301 cgtccatccc accaccaagc ccggcgaact tggcaactgc gtgttcgcgt tgtactacga
      361 acatttcatt gtcactttcc tctaaccttt ccaggcggtg atcaacggca tggataccgg
      421 gcatcttgat gttcccgacg gtcttttttg acctgtaagt tgttacgaaa gagcaagaca
      481 gatggccacc accatcaact ttcagtgcca tatgagtgta tcccctcaga ccaccatctg
      541 ctggatacat catctctgta ttaggctccc aacccttggt cttcttctgc atcacgggac
      601 cattggaggg aaagtttacc cctctgactt ggacgtggta aacgagacag ccatcctcaa
      661 ggctggtgtc ctgcatgacg gtgacgactc caccatcttc gtatctcgta actctttccc
      721 aagtaaaacc ctcaggaaag gactgtttaa agaaatcagg aatgcctttc gggtacttga
      781 taaaagtacg gctgccatac atgaacgacg tggcaagaat gtcaaaggca aatggcaggg
      841 gtcctccctc gatgactttg atcctcatgg tttgagttcc catgtacgga ttgccttctc
      901 cttcacctgt gcatttgaat tggtggccgt tgaccgaacc ttccatgacc acctgtgaaa
      961 aaaaaaaaga agactcgaag ttagtacaat tgaaaaaagt gatttaattt ataccatttt
     1021 aattcagctt tgtaaaaatg tatcaaagag atagcaaggt attcagtttt agtaaacaag
     1081 ataattgctc ctaaagtagc cccttgaatt ctagcccctt gaagtccgag gcagtaggca
     1141 ccgcttgaag tctttaatta aatacatctg tggcttcact atttaattaa agacttcaag
     1201 cggcgctcac tgtcaacagc aatatacctt ctcgagcctt ctgttgggtt aagaagtccc
     1261 tgaagaagta atcccagcaa gtgtttccaa gatgtgcagg caacgattct gtaaagtact
     1321 gaagcctcat tcaacatagt atatgtgctg ccgaagcgag cacacttacc ttcatacgca
     1381 tattttcctt gatcagctct tcgcccttag acaccatagg tccagggttc tcctccacgt
     1441 ctccagcctg cttcagcagg ctgaagttag tagctccgct tccagcatct tttccggaat
     1501 aggccaagga agacaagtcc agtttggttt ttaaggcttt agtgagctct gcggccagca
     1561 gctcatggag ggacacgccg tcgtgggagt acacccagtt tttcccagtc cagtcataac
     1621 gcttaggtcc actggatgga gaagatagcc agatttgctt gtttggcgtc tgcttgttga
     1681 tcacataggt tcctagatct ccacccagtt tgacagttaa gacaccactc ccaaaggaga
     1741 catcatagtc ctcaaacgtg tatggcttgt ctgcaaggtc ttcaaaaaac tctgctaaag
     1801 agtccagcgt ttcctctgct agtctttcat aggtggtctc atctagagag cctgggtggc
     1861 ccaaagttcc agatttcctc aaattcatca aatagacact ctgctttttg acattccaaa
     1921 tctggttgag gccacgttgg ttcgaacttg cgcggcgggg cgtgcaggtc gcatcgatgt
     1981 cggtgcgcag gccacggcgg ccgcagagtg gggccaactc tgccggccgc gggacccggg
     2041 tgagggtctg ggcctgggcc gggctgggtg acgccaggag gccggctact gcgcggcgcc
     2101 cgagagtcca catggtggcc attacagcgt ggatggcgtc tccaggcgat ctgacggttc
     2161 actaaacgag ctctttacga gggtaggaag tggtacggaa agttggtata agacaaaagt
     2221 gttgtggaat tgctccaggc gatctgacgg ttcactaaac gagctctgct tttataggcg
     2281 cccaccgtac acgcctaaag cttatacgtt ctctatcact gatagggagt aaactggata
     2341 tacgttctct atcactgata gggagtaaac tgtagatacg ttctctatca ctgataggga
     2401 gtaaactggt catacgttct ctatcactga tagggagtaa actccttata cgttctctat
     2461 cactgatagg gagtaaagtc tgcatacgtt ctctatcact gatagggagt aaactcttca
     2521 tacgttctct atcactgata gggagtaaac tcgaggaata gtacgaacat ggtagctagt
     2581 ggatccttcg tcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc
     2641 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
     2701 cttccccgat aacggagacc ggcacactgg cgcatatcgg tggtcatcat gcgccagctt
     2761 tcatccccga tatgcaccac cgggtaaagt tcacgcatat gaagctggac atcacctccc
     2821 acaacctcga ggtggaacag tacgctgata tctcttatac tgggcagagc gcacatcgcc
     2881 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     2941 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     3001 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     3061 cgccagaaca cagtaatgtg ccaccatgag ccggctggac aagagcaaag tgatcaacag
     3121 cgccctggaa ctgctgaacg gcgtgggcat cgagggcctg accacccgga agctggccca
     3181 gaaactgggc gtggaacagc ccaccctgta ctggcacgtg aagaacaagc gggccctgct
     3241 ggacgccctg cccatcgaga tgctggaccg gcaccacacc cacagctgcc ctctggaagg
     3301 cgagagctgg caggacttcc tgcggaacaa cgccaagagc tacagatgcg ccctgctgag
     3361 ccaccgggac ggcgccaaag tgcacctggg caccagaccc accgagaagc agtacgagac
     3421 actggaaaac cagctggcct tcctgtgcca gcagggcttc agcctggaaa acgccctgta
     3481 cgccctgagc gccgtgggcc acttcaccct gggctgcgtg ctggaagaac aggaacacca
     3541 ggtcgccaaa gaggaaagag agacacccac caccgacagc atgccccccc tgctgaagca
     3601 ggccatcgag ctgttcgaca gacagggcgc cgagcccgcc ttcctgttcg gcctggaact
     3661 gatcatctgc ggcctcgaga agcagctgaa gtgcgagagc ggcggaccca ccgacgccct
     3721 ggacgacttc gacctggaca tgctgcccgc cgatgccctg gatgattttg atctggatat
     3781 gctccctgct gacgctctcg atgacttcga tctcgatatg ctgccaggcg gaagcggagc
     3841 tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg gacctatggt
     3901 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     3961 cgtaaacggc cacaagttca gcgtccgcgg cgagggcgag ggcgatgcca ccaacggcaa
     4021 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     4081 gaccacctta ggctacggcg tggcctgctt cgcccgctac cccgaccaca tgaagcagca
     4141 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tctctttcaa
     4201 ggacgacggt acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     4261 ccgcatcgtg ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     4321 ggagtacaac ttcaacagcc acaaggtcta tatcacggcc gacaagcaga agaacggcat
     4381 caaggctaac ttcaagaccc gccacaacgt tgaggacggc ggcgtgcagc tcgccgacca
     4441 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     4501 gagccatcag tccaaactga gcaaagaccc caacgagaag cgcgatcaca tggtcctgaa
     4561 ggagagggtg accgccgccg ggattacaca tgacatggac gagctgtaca agtaacaacg
     4621 ccgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4681 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4741 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4801 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4861 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4921 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4981 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     5041 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     5101 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5161 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgccgctcg
     5221 ctgatatagg gcccggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5281 cactttttaa aagaaaaggg gggactggaa gggctagctc actcccaacg aagacaagat
     5341 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5401 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5461 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5521 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     5581 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     5641 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5701 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     5761 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5821 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5881 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     5941 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6001 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6061 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     6121 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6181 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6241 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6301 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6361 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6421 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6481 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6541 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     6601 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6661 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6721 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6781 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6841 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6901 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6961 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7021 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7081 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7141 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7201 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     7261 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     7321 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     7381 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7441 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7501 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7561 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7621 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7681 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7741 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7801 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7861 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7921 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7981 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8041 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8101 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8161 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     8221 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     8281 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     8341 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     8401 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8461 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     8521 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     8581 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     8641 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     8701 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     8761 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     8821 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     8881 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     8941 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     9001 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     9061 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     9121 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     9181 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     9241 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     9301 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     9361 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     9421 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     9481 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     9541 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     9601 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     9661 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     9721 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     9781 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     9841 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     9901 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     9961 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
    10021 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
    10081 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
    10141 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
    10201 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
    10261 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
    10321 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
    10381 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
    10441 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
    10501 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
    10561 tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt ggggggtaca
    10621 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
    10681 aacaaattac aaaattcaaa attttatcgt ccagtgtggt ggc
//
