LOCUS       sequence-456375-       12265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Second generation lenti tetracycline operator (on).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12265)
  AUTHORS   Mertens J, Paquola AC, Ku M, Hatch E, Bohnke L, Ladjevardi S, 
            McGrath S, Campbell B, Lee H, Herdy JR, Goncalves JT, Toda T, Kim Y,
            Winkler J, Yao J, Hetzer MW, Gage FH
  TITLE     Directly Reprogrammed Human Neurons Retain Aging-Associated 
            Transcriptomic Signatures and Reveal Age-Related Nucleocytoplasmic 
            Defects.
  JOURNAL   Cell Stem Cell. 2015 Oct 6. pii: S1934-5909(15)00408-7. doi: 
            10.1016/j.stem.2015.09.001.
  PUBMED    26456686
REFERENCE   2  (bases 1 to 12265)
  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 Stem 
            Cell. 2015 Oct 6. pii: S1934-5909(15)00408-7. doi: 
            10.1016/j.stem.2015.09.001."
COMMENT     Sequence Label: pLVX-EF1a-tetOn-IRES-G418 (EtO)
FEATURES             Location/Qualifiers
     source          1..12265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    680..805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1302..1535
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1720..1764
                     /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    2027..2142
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2282..3460
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2513..3451
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3408..3428
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(3483..4100,4101..4106,4107..4142,4143..4145,4146..4181,
                     4182..4184,4185..4220,4221..4229)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    join(4256..4830,4831..4833,4834..4842)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4423..4440)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4650..4669
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4843..5637
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4897..4916)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5507..5526
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    5833..6421
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5886..5906)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6304..6315)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6628..7261
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7368..7385)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7374..7598
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7644..8072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7731..7750)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7941..7962
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8081..8101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8086..8415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8266..8401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8328..8347
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8464..9489
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             8464..9489
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    9619..9740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9789..9805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9789..9805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9802..9824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9813..9829
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9837..9843,9844..9861,9862..9867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9882..9903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10020..10037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10191..10779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10271..10290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10950..11741,11742..11810))
                     /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     11573..11592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11811..11915)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    11963..12097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             join(12265,1..633)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
       61 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
      121 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
      181 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
      241 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
      301 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
      361 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
      421 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
      481 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
      541 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
      601 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
      661 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
      721 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
      781 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
      841 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
      901 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
      961 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1021 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1081 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1141 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1201 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1261 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1321 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1381 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1441 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1501 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1561 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1621 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1681 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1741 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1801 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1861 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1921 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1981 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccttta aaagaaaagg
     2041 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2101 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2161 cagcagagat ccagtttatc gatgcgcccg acgataagct ttgcaaagat ggataaagtt
     2221 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
     2281 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     2341 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     2401 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     2461 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     2521 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     2581 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     2641 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     2701 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     2761 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     2821 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     2881 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     2941 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     3001 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     3061 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     3121 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     3181 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     3241 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     3301 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     3361 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     3421 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggaatttcga cccggatcca
     3481 ccatgtctag actggacaag agcaaagtca taaacggcgc tctggaatta ctcaatggag
     3541 tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta
     3601 ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca atcgagatgc
     3661 tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc
     3721 ggaacaacgc caagtcattc cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc
     3781 atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc
     3841 tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact
     3901 ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag gaaagagaga
     3961 cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccggc
     4021 agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac
     4081 agctaaagtg cgaaagcggc gggccggccg acgcccttga cgattttgac ttagacatgc
     4141 tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg
     4201 attttgacct tgacatgctc cccgggtaac tagaattccg cccccccccc ccccccccct
     4261 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     4321 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     4381 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     4441 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     4501 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     4561 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     4621 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     4681 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     4741 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     4801 tggttttcct ttgaaaaaca cgatgataag cttgccacaa ccatggctga acaagatgga
     4861 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4921 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4981 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5041 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5101 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5161 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5221 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5281 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5341 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5401 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5461 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5521 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5581 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5641 ggatcggcta gagtcgaccc ggacgggacg ctcctgcgcc tgatacagaa cgaattgctt
     5701 gcaggcatct catgagtgtg tcttcccgtt ttccgcctga ggtcactgcg tggatggagc
     5761 gctggcgcct gctgcgcgac ggcgagctgc tcaccaccca ctcgagtcta gagggcccga
     5821 cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5881 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5941 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6001 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6061 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6121 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6181 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     6241 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6301 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     6361 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     6421 cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata
     6481 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     6541 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     6601 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
     6661 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     6721 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     6781 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     6841 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     6901 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     6961 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     7021 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     7081 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     7141 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     7201 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     7261 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     7321 gagagtgaga ggccttgaca ttgctagcgt ttaaacccgc tgatcagcct cgactgtgcc
     7381 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     7441 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     7501 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     7561 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     7621 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     7681 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     7741 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7801 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     7861 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7921 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     7981 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     8041 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     8101 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     8161 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     8221 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     8281 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     8341 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     8401 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     8461 gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     8521 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     8581 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     8641 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     8701 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     8761 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     8821 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     8881 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     8941 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     9001 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     9061 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     9121 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     9181 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     9241 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     9301 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     9361 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     9421 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     9481 aaggaatagc acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     9541 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     9601 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     9661 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     9721 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     9781 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     9841 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     9901 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     9961 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    10021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    10081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    10141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    10201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    10261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    10321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    10381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    10441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    10501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    10561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    10621 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    10681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    10741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    10801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    10861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    10921 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    10981 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    11041 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    11101 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    11161 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    11221 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    11281 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    11341 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    11401 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    11461 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    11521 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    11581 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    11641 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    11701 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    11761 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    11821 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    11881 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    11941 gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag
    12001 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    12061 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
    12121 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
    12181 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta
    12241 aatatgtact acaaacttag tagtt
//
