LOCUS       sequence-472782-       10264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of ComMAND base gene regulating FXN-P2A-mRuby2
            (therapeutically relevant gene).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10264)
  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 10264)
  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-bGH]-EFS-rtTA-P2A-mGL-WPRE
FEATURES             Location/Qualifiers
     source          1..10264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             379..423
                     /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    690..806
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    833..1057
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(1072..1782)
                     /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"
     CDS             complement(1783..1839)
                     /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(2539..2917)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2675..2693
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2711..2729
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2747..2765
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2783..2801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2819..2837
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2855..2873
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2891..2909
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(3098..3117)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        3227..3438
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3451..4194
                     /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             4204..4260
                     /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(4261..4263,4264..4266,4267..4980)
                     /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(4306..4327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    4991..5579
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5044..5064)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5462..5473)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5671..5904
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5976..6097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6013..6032)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6067..6086
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6137..6272
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6292..6311)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6293..6311)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6321..6338)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6321..6337)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6330..6352)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6479..6934
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6566..6585)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6776..6797
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6960..7064
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7065..7133,7134..7925)
                     /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(7283..7302)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8096..8684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8585..8604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8838..8855
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8972..8993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9008..9013,9014..9031,9032..9038)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9046..9062
                     /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     9051..9073
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9070..9086
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9070..9086
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9105..9125
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9107..9125
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9153..9379
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9380..9560
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9607..9732
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    join(10225..10264,1..194)
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
       61 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
      121 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
      181 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
      241 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
      301 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
      361 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
      421 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
      481 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
      541 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
      601 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
      661 gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac
      721 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
      781 aaacaaatta caaaattcaa aattttatcg tccagtgtgg tggctactag cgccatagag
      841 cccaccgcat ccccagcatg cctgctattg tcttcccaat cctccccctt gctgtcctgc
      901 cccaccccac cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca
      961 ttttattagg aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag
     1021 gggcaaacaa cagatggctg gcaactagaa ggcacaggtg caacgttgtt acttgtacag
     1081 ctcgtccatc ccaccaccaa gcccggcgaa cttggcaact gcgtgttcgc gttgtactac
     1141 gaacatttca ttgtcacttt cctctaacct ttccaggcgg tgatcaacgg catggatacc
     1201 gggcatcttg atgttcccga cggtcttttt tgacctgtaa gttgttacga aagagcaaga
     1261 cagatggcca ccaccatcaa ctttcagtgc catatgagtg tatcccctca gaccaccatc
     1321 tgctggatac atcatctctg tattaggctc ccaacccttg gtcttcttct gcatcacggg
     1381 accattggag ggaaagttta cccctctgac ttggacgtgg taaacgagac agccatcctc
     1441 aaggctggtg tcctgcatga cggtgacgac tccaccatct tcgtatctcg taactctttc
     1501 ccaagtaaaa ccctcaggaa aggactgttt aaagaaatca ggaatgcctt tcgggtactt
     1561 gataaaagta cggctgccat acatgaacga cgtggcaaga atgtcaaagg caaatggcag
     1621 gggtcctccc tcgatgactt tgatcctcat ggtttgagtt cccatgtacg gattgccttc
     1681 tccttcacct gtgcatttga attggtggcc gttgaccgaa ccttccatga ccaccttcat
     1741 acgcatattt tccttgatca gctcttcgcc cttagacacc ataggtccag ggttctcctc
     1801 cacgtctcca gcctgcttca gcaggctgaa gttagtagct ccgcttccag catcttttcc
     1861 ggaataggcc aaggaagaca agtccagttt ggtttttaag gctttagtga gctctgcggc
     1921 cagcagctca tggagggaca cgccgtcgtg ggagtacacc cagtttttcc cagtccagtc
     1981 ataacgctta ggtccactgg atggagaaga tagccagatt tgcttgtttg gcgtctgctt
     2041 gttgatcaca taggttccta gatctccacc cagtttgaca gttaagacac cactcccaaa
     2101 ggagacatca tagtcctcaa acgtgtatgg cttgtctgca aggtcttcaa aaaactctgc
     2161 taaagagtcc agcgtttcct ctgctagtct ttcataggtg gtctcatcta gagagcctgg
     2221 gtggcccaaa gttccagatt tcctcaaatt catcaaatag acactctgct ttttgacatt
     2281 ccaaatctgg ttgaggccac gttggttcga acttgcgcgg cggggcgtgc aggtcgcatc
     2341 gatgtcggtg cgcaggccac ggcggccgca gagtggggcc aactctgccg gccgcgggac
     2401 ccgggtgagg gtctgggcct gggccgggct gggtgacgcc aggaggccgg ctactgcgcg
     2461 gcgcccgaga gtccacatgg tggccattac agcgtggatg gcgtctccag gcgatctgac
     2521 ggttcactaa acgagctctt tacgagggta ggaagtggta cggaaagttg gtataagaca
     2581 aaagtgttgt ggaattgctc caggcgatct gacggttcac taaacgagct ctgcttttat
     2641 aggcgcccac cgtacacgcc taaagcttat acgttctcta tcactgatag ggagtaaact
     2701 ggatatacgt tctctatcac tgatagggag taaactgtag atacgttctc tatcactgat
     2761 agggagtaaa ctggtcatac gttctctatc actgataggg agtaaactcc ttatacgttc
     2821 tctatcactg atagggagta aagtctgcat acgttctcta tcactgatag ggagtaaact
     2881 cttcatacgt tctctatcac tgatagggag taaactcgag gaatagtacg aacatggtag
     2941 ctagtggatc cttcgtcgac ctgcagactg gctgtgtata agggagcctg acatttatat
     3001 tccccagaac atcaggttaa tggcgttttt gatgtcattt tcgcggtggc tgagatcagc
     3061 cacttcttcc ccgataacgg agaccggcac actggcgcat atcggtggtc atcatgcgcc
     3121 agctttcatc cccgatatgc accaccgggt aaagttcacg catatgaagc tggacatcac
     3181 ctcccacaac ctcgaggtgg aacagtacgc tgatatctct tatactgggc agagcgcaca
     3241 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     3301 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     3361 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     3421 tttgccgcca gaacacagta atgtgccacc atgagccggc tggacaagag caaagtgatc
     3481 aacagcgccc tggaactgct gaacggcgtg ggcatcgagg gcctgaccac ccggaagctg
     3541 gcccagaaac tgggcgtgga acagcccacc ctgtactggc acgtgaagaa caagcgggcc
     3601 ctgctggacg ccctgcccat cgagatgctg gaccggcacc acacccacag ctgccctctg
     3661 gaaggcgaga gctggcagga cttcctgcgg aacaacgcca agagctacag atgcgccctg
     3721 ctgagccacc gggacggcgc caaagtgcac ctgggcacca gacccaccga gaagcagtac
     3781 gagacactgg aaaaccagct ggccttcctg tgccagcagg gcttcagcct ggaaaacgcc
     3841 ctgtacgccc tgagcgccgt gggccacttc accctgggct gcgtgctgga agaacaggaa
     3901 caccaggtcg ccaaagagga aagagagaca cccaccaccg acagcatgcc ccccctgctg
     3961 aagcaggcca tcgagctgtt cgacagacag ggcgccgagc ccgccttcct gttcggcctg
     4021 gaactgatca tctgcggcct cgagaagcag ctgaagtgcg agagcggcgg acccaccgac
     4081 gccctggacg acttcgacct ggacatgctg cccgccgatg ccctggatga ttttgatctg
     4141 gatatgctcc ctgctgacgc tctcgatgac ttcgatctcg atatgctgcc aggcggaagc
     4201 ggagctacta acttcagcct gctgaagcag gctggagacg tggaggagaa ccctggacct
     4261 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4321 ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac
     4381 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4441 ctcgtgacca ccttaggcta cggcgtggcc tgcttcgccc gctaccccga ccacatgaag
     4501 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct
     4561 ttcaaggacg acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4621 gtgaaccgca tcgtgctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4681 aagctggagt acaacttcaa cagccacaag gtctatatca cggccgacaa gcagaagaac
     4741 ggcatcaagg ctaacttcaa gacccgccac aacgttgagg acggcggcgt gcagctcgcc
     4801 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     4861 tacctgagcc atcagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4921 ctgaaggaga gggtgaccgc cgccgggatt acacatgaca tggacgagct gtacaagtaa
     4981 caacgccgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5041 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5101 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5161 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5221 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5281 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5341 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     5401 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5461 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5521 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5581 gctcgctgat atagggcccg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5641 ttagccactt tttaaaagaa aaggggggac tggaagggct agctcactcc caacgaagac
     5701 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5761 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5821 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5881 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     5941 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6001 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6061 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6121 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6181 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     6241 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     6301 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     6361 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     6421 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     6481 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6541 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6601 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6661 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6721 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6781 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     6841 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     6901 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     6961 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7021 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     7081 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     7141 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7201 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7261 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7321 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7381 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7441 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7501 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7561 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7621 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7681 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7741 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7801 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7861 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7921 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7981 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     8041 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     8101 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     8161 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8221 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8281 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8341 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8401 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8461 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8521 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8581 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8641 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8701 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8761 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8821 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     8881 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     8941 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     9001 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     9061 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     9121 aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc
     9181 ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc
     9241 atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg
     9301 tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg
     9361 cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc
     9421 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     9481 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     9541 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     9601 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     9661 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     9721 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     9781 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     9841 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     9901 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     9961 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
    10021 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
    10081 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
    10141 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
    10201 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
    10261 aagc
//
