LOCUS       sequence-6996-ex       11876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-regulated (Tet-on) lentiviral vector for shGATA1 (hUbiquitin 
            promoter) - 3rd generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11876)
  AUTHORS   Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P
  TITLE     A versatile tool for conditional gene expression and knockdown.
  JOURNAL   Nat Methods. 2006 Feb;3(2):109-16. doi: 10.1038/nmeth846
  PUBMED    16432520
REFERENCE   2  (bases 1 to 11876)
  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"; doi: "10"; journalName: 
            "Nat Methods"; date: "2006-02"; volume: "3"; issue: "2"; pages: 
            "109-16"
COMMENT     Sequence Label: pLVUTHshGATA1-tTR-KRAB
FEATURES             Location/Qualifiers
     source          1..11876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2622..3833
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3670..3689
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3854..3863
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3860..3862,3863..3865,3866..4579)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3860..3862,3863..3865,3866..4576)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3905..3926)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4166..4185)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4513..4534
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4618..5185
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4636..5187
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4780..4797)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5007..5026
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5190..5810
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MARLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(5263..5282)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             5811..5831
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5934..6128
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     misc_feature    6268..6856
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6321..6341)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6739..6750)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6859..6875)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6860..6876)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    join(7026..7044,7045..7067,7068..7086,7087..7109,7110..7128,
                     7129..7151,7152..7170,7171..7193,7194..7212,7213..7235,
                     7236..7254,7255..7277,7278..7296)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        7325..7539
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     7460..7479
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    complement(7630..7663)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             7683..7863
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7887..7904)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7893..8117
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8163..8591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8250..8269)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8460..8481
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8600..8620)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8605..8934
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8785..8920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8847..8866
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8982..9029
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9048..9422
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    9552..9673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9589..9608)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9643..9662
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9722..9738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9722..9738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9735..9757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9746..9762
                     /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(9770..9776,9777..9794,9795..9800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9815..9836
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9953..9970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10124..10712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10204..10223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10883..11674,11675..11743))
                     /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     11506..11525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11744..11848)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta acccgtgtcg gctccagatc tggcctccgc gccgggtttt
     2641 ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg
     2701 cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2761 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2821 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2881 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2941 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3001 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3061 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3121 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3181 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     3241 ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc
     3301 ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg
     3361 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     3421 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     3481 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     3541 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     3601 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     3661 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     3721 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     3781 agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacaggatcc
     3841 ccgggtaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3901 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3961 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4021 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4081 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4141 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4201 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4261 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4321 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     4381 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4441 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4501 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4561 ggacgagctg tacaagtaaa gcggccgcga ctctagaatt cgatgggctg caggaattcc
     4621 gccccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     4681 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     4741 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     4801 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     4861 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     4921 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     4981 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     5041 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     5101 ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     5161 tttcctttga aaaacacgat gataatacca tggctagatt agataaaagt aaagtgatta
     5221 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     5281 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     5341 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     5401 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     5461 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     5521 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     5581 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     5641 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     5701 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     5761 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtcg ccaaaaaaga
     5821 agagaaaggt cgacggcggt ggtgctttgt ctcctcagca ctctgctgtc actcaaggaa
     5881 gtatcatcaa gaacaaggag ggcatggatg ctaagtcact aactgcctgg tcccggacac
     5941 tggtgacctt caaggatgta tttgtggact tcaccaggga ggagtggaag ctgctggaca
     6001 ctgctcagca gatcgtgtac agaaatgtga tgctggagaa ctataagaac ctggtttcct
     6061 tgggttatca gcttactaag ccagatgtga tcctccggtt ggagaaggga gaagagccct
     6121 ggctggtgga gagagaaatt caccaagaga cccatcctga ttcagagact gcatttgaaa
     6181 tcaaatcatc agtttaagga tctcgactag ctagtagcta gctagctagt cgagctcaac
     6241 ttcgaattcg atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga
     6301 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6361 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6421 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6481 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6541 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6601 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6661 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6721 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6781 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6841 tgggccgcct ccccgcatcg ataccgtcga cctcgagtac ctttaagacc aatgacttac
     6901 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     6961 cactcccaac gaagacaaga tgggatcaat tcaccatgct agtggatccc cctcgagttt
     7021 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
     7081 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
     7141 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
     7201 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     7261 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcgg ggctgcagga
     7321 attcgaacgc tgacgtcatc aacccgctcc aaggaatcgc gggcccagtg tcactaggcg
     7381 ggaacaccca gcgcgcgtgc gccctggcag gaagatggct gtgagggaca ggggagtggc
     7441 gccctgcaat atttgcatgt cgctatgtgt tctgggaaat caccataaac gtgaaatgtc
     7501 tttggatttg ggaatcttat aagttctgta tgagaccaca gatccccgaa gcgcctgatt
     7561 gtcagtttca agagaactga caatcaggcg cttctttttg gaaaagctta tcgataccgt
     7621 cgcatgggaa taacttcgta tagcatacat tatacgaagt tatgctgctt tttgcttgta
     7681 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     7741 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     7801 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7861 gcagcatcta gaaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     7921 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     7981 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     8041 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     8101 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     8161 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     8221 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     8281 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     8341 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     8401 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     8461 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     8521 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     8581 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     8641 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     8701 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     8761 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     8821 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     8881 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     8941 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     9001 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     9061 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     9121 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     9181 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     9241 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     9301 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     9361 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     9421 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     9481 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     9541 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     9601 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     9661 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     9721 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     9781 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     9841 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     9901 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     9961 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
    10021 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
    10081 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
    10141 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
    10201 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
    10261 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
    10321 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
    10381 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
    10441 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
    10501 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
    10561 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
    10621 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
    10681 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
    10741 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
    10801 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
    10861 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
    10921 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
    10981 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
    11041 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
    11101 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
    11161 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
    11221 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
    11281 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
    11341 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
    11401 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    11461 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    11521 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    11581 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    11641 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    11701 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    11761 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    11821 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
