LOCUS       sequence-292008-       13460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox inducible expression of luciferase + crIFNg. Constitutive EF1a 
            driven rtTA and GFP .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13460)
  AUTHORS   Kempton HR, Goudy LE, Love KS, Qi LS
  TITLE     Multiple Input Sensing and Signal Integration Using a Split Cas12a 
            System.
  JOURNAL   Mol Cell. 2020 Jan 30. pii: S1097-2765(20)30037-X. doi: 
            10.1016/j.molcel.2020.01.016.
  PUBMED    32027839
REFERENCE   2  (bases 1 to 13460)
  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: "Mol Cell. 
            2020 Jan 30. pii: S1097-2765(20)30037-X. doi: 
            10.1016/j.molcel.2020.01.016."
COMMENT     Sequence Label: pSLQ8703
            pHR-R(TRE3G-Fluc-2XcrIFNg-SV40pA)-EF1...
FEATURES             Location/Qualifiers
     source          1..13460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     202..222
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(292..309)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(463..1051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(543..562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1222..2013,2014..2082))
                     /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     1845..1864
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2083..2187)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2255..2273
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     2433..2452
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2666..2685
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2975..2993
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2975..2992
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     LTR             3018..3251
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     3341..3357
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3342..3358
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3361..3949
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3467..3478
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3876..3896
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4009..4755)
                     /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             complement(4768..4821)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(join(4831..5541,5542..5544))
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             complement(4831..5544)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4871..4891)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     5486..5514
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     intron          5569..6491
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5592..5612)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    6686..6704
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6722..6740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6758..6776
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6794..6812
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6830..6848
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6866..6884
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6902..6920
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6965..6989
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             7066..8718
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(7132..7150)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    9106..9223
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(9508..9552)
                     /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    9737..9970
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    10467..10592
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             10639..11272
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    11440..11574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(11999..12019)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          12149..12214
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     promoter        complement(join(13348..13460,1..217))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(13362..13460,1..37)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
       61 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
      121 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
      181 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgc attaatgaat
      241 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      301 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      361 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      421 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      481 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      541 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      601 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      661 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      721 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      781 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      841 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      901 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      961 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1021 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1081 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1141 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1201 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1261 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1321 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1381 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1441 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1501 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1561 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1621 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1681 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1741 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1801 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1861 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1921 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1981 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     2041 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2101 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2161 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     2221 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     2281 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     2341 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     2401 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     2461 gcaccattcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     2521 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     2581 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     2641 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     2701 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgg ctagcgatga
     2761 ccctgctgat tggttcgctg accatttccg ggtgcgggac ggcgttacca gaaactcaga
     2821 aggttcgtcc aaccaaaccg actctgacgg cagtttacga gagagatgat agggtctgct
     2881 tcagtaagcc agatgctaca caattaggct tgtacatatt gtcgttagaa cgcggctaca
     2941 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaatacacg
     3001 gaattaattc tagatgctgc tagagatttt ccacactgac taaaagggtc tgagggatct
     3061 ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct
     3121 ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg
     3181 gtctaaccag agagacccag tacaagcaaa aagcagatct tgtcttcgtt gggagtgaat
     3241 tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag ctgccttgta
     3301 agtcattggt cttaaaggta ccgagctcga attaattccc tcgaggtcga cggtatcgat
     3361 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
     3421 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
     3481 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg agcagccaag
     3541 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
     3601 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
     3661 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
     3721 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
     3781 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     3841 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     3901 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tcgataagct
     3961 tgatatcaag cttgcatgcc tgcaggtcga ctctagagtc gcggccgctt agcctggcag
     4021 catatcgaga tcgaagtcat cgagagcgtc agcagggagc atatccagat caaaatcatc
     4081 cagggcatcg gcgggcagca tgtccaggtc gaagtcgtcc agggcgtcgg tgggtccgcc
     4141 gctctcgcac ttcagctgct tctcgaggcc gcagatgatc agttccaggc cgaacaggaa
     4201 ggcgggctcg gcgccctgtc tgtcgaacag ctcgatggcc tgcttcagca gggggggcat
     4261 gctgtcggtg gtgggtgtct ctctttcctc tttggcgacc tggtgttcct gttcttccag
     4321 cacgcagccc agggtgaagt ggcccacggc gctcagggcg tacagggcgt tttccaggct
     4381 gaagccctgc tggcacagga aggccagctg gttttccagt gtctcgtact gcttctcggt
     4441 gggtctggtg cccaggtgca ctttggcgcc gtcccggtgg ctcagcaggg cgcatctgta
     4501 gctcttggcg ttgttccgca ggaagtcctg ccagctctcg ccttccagag ggcagctgtg
     4561 ggtgtggtgc cggtccagca tctcgatggg cagggcgtcc agcagggccc gcttgttctt
     4621 cacgtgccag tacagggtgg gctgttccac gcccagtttc tgggccagct tccgggtggt
     4681 caggccctcg atgcccacgc cgttcagcag ttccagggcg ctgttgatca ctttgctctt
     4741 gtccagccgg ctcatcgacc ctccgcctgg gccaggattc tcctcgacgt caccgcatgt
     4801 tagcagactt cctctgccct cggaaccgcc tttgtagagc tcatccatgc catgtgtaat
     4861 cccagcagca gttacaaact caagaaggac catgtggtca cgcttttcgt tgggatcttt
     4921 cgaaaggaca gattgtgtcg acaggtaatg gttgtctggt aaaaggacag ggccatcgcc
     4981 aattggagta ttttgttgat aatggtctgc tagttgaacg gaaccatctt caacgttgtg
     5041 gcgaattttg aagttagctt tgattccatt cttttgtttg tctgccgtga tgtatacatt
     5101 gtgtgagtta aagttgtact cgagtttgtg tccaagaatg tttccatctt ctttaaaatc
     5161 aatacccttt aactcgatac gattaacaag ggtatcacct tcaaacttga cttcagcacg
     5221 cgtcttgtag gtcccgtcat ctttgaaaga tatagtgcgt tcctgtacat aaccttcggg
     5281 catggcactc ttgaaaaagt catgccgttt catgtgatcc ggataacggg aaaagcattg
     5341 aacaccatag gtcagagtag tgacaagtgt tggccacgga acaggtagtt ttccagtagt
     5401 gcaaataaat ttaagggtga gttttccgtt tgtagcatca ccttcaccct ctccacggac
     5461 agaaaatttg tgcccattaa catcaccatc taattcaaca agaattggga caactccagt
     5521 gaaaagttct tctcctttgc tcatggtcgt acgcgttgtt cacgacacct gaaatggaag
     5581 aaaaaaactt tgaaccactg tctgaggctt gagaatgaac caagatccaa actcaaaaag
     5641 gccaaattcc aaggagaatt acatcaagtg ccaagctggc ctaacttcag tctccaccca
     5701 ctcagtgtgg ggaaactcca tcgcataaaa cccctccccc caacctaaag acgacgtact
     5761 ccaaaagctc cagaactaat cgaggtgcct ggacggcgcc cggtactccg tggagtcaca
     5821 tgaagcgacg gctgaggacg gaaaggccct tttcctttgt gtgggtgact cacccgcccg
     5881 ctctcccgag cgccgcgtcc tccattttga gccccctgga gcagggccgg gaagcggcca
     5941 tctttccgct cacgcaactg gtgccgaccg ggccagcctt gccgcccagg gcggggcgat
     6001 acacggcggc gcgaggccag gcaccagagc aggccggcca gcttgagact acccccgtcc
     6061 gattctcggt ggccgcgctc gcaggccccg cctcgccgaa catgtgcgct gggacgcacg
     6121 ggccccgtcg ccggccgcgg gcccaaaaac cgaaatacca gtgtgcagat cctggcccgc
     6181 atttacaaga ctatcttgcc agaaaaaaag cgtcgcagca cgtcatcaaa aattttaaat
     6241 ggctagagac ttatcgaaag cagcgagaca ggcgcgaagg tgccaccaga ttcgcacgcg
     6301 gcggccccag cgcccaggcc aggcctcaac tcaagcacga ggcgaagggg ctcctaaagc
     6361 gcaaggcccg cccctggctc cagctcggga tcaagaatca cgtactggag ccaggtggaa
     6421 gtaattcaag gcacgcaagg gccataaccc gtaaagaggc caggcccgcg ggaaccacac
     6481 acggcactta cctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact
     6541 actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca
     6601 tcactttccc agtttacccc gcgccacctt ctctaggcac cggttcaatt gcatatgctg
     6661 atatctccac cccacagtgg tttactccct atcagtgata gagaacgtat gaagagttta
     6721 ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga tagagaacgt
     6781 ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact ccctatcagt
     6841 gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat atccagttta
     6901 ctccctatca gtgatagaga acgtataagc tttaggcgtg tacggtgggc gcctataaaa
     6961 gcagagctcg tttagtgaac cgtcagatcg cctggagcaa ttccacaaca cttttgtctt
     7021 ataccaactt tccgtaccac ttcctaccct cgtaaagaat tcaccatgga agacgccaaa
     7081 aacataaaga aaggcccggc gccattctat ccactagagg atggaaccgc tggagagcaa
     7141 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
     7201 catatcgagg tgaacatcac gtacgcggaa tacttcgaaa tgtccgttcg gttggcagaa
     7261 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
     7321 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     7381 aacgacattt ataatgaacg tgaattgctc aacagtatga acatttcgca gcctaccgta
     7441 gtgtttgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa attaccaata
     7501 atccagaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     7561 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt accagagtcc
     7621 tttgatcgtg acaaaacaat tgcactgata atgaactcct ctggatctac tgggttacct
     7681 aagggtgtgg cccttccgca tagaactgcc tgcgtcagat tctcgcatgc cagagatcct
     7741 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     7801 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     7861 tatagatttg aagaagagct gtttttacga tcccttcagg attacaaaat tcaaagtgcg
     7921 ttgctagtac caaccctatt ttcattcttc gccaaaagca ctctgattga caaatacgat
     7981 ttatctaatt tacacgaaat tgcttctggg ggcgcacctc tttcgaaaga agtcggggag
     8041 gcggttgcaa aacgcttcca tcttccaggg atacgacaag gatatgggct cactgagact
     8101 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     8161 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     8221 cagagaggcg aattatgtgt cagaggacct atgattatgt ccggttatgt aaacaatccg
     8281 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     8341 tgggacgaag acgaacactt cttcatagtt gaccgcttga agtctttaat taaatacaaa
     8401 ggataccagg tggcccccgc tgaattggag tcgatattgt tacaacaccc caacatcttc
     8461 gacgcgggcg tggcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     8521 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     8581 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     8641 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     8701 ggaaagtcca aattgtaagg atcctttctt tctttgaatt tctactaagt gtagatagat
     8761 gagatggtga cagataggca atttctacta agtgtagata gatgagatgg tgacagatag
     8821 gcaatttcta ctaagtgtag atcgagtttt ctttttgttt tgtacaagta agttacgcgt
     8881 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     8941 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     9001 tcatgtctgg atcaactgga taactcaagc taaccgtcat actagtaatt cgatatcaag
     9061 cttatcgatc caaactggat ctctgctgtc cctgtaataa acccgaaaat tttgaatttt
     9121 tgtaatttgt ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct
     9181 ttcccctgca ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc
     9241 catttggcga taccgtcgag atccgttcac taatcgaatg gatctgtctc tgtctctctc
     9301 tccaccttct tcttctattc cttcgggcct gtcgggtccc ctcggggttg ggaggtgggt
     9361 ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa gtacagcaaa
     9421 aactattctt aaacctacca agcctcctac tatcattatg aataatttta tataccacag
     9481 ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt ccaataattc
     9541 ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta ttaagcttgt
     9601 gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa tctgttccag
     9661 agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga gttttccaga
     9721 gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag ccaggattct
     9781 tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt gcgcctcaat
     9841 agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc tggcctgtac
     9901 cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga acccaaggaa
     9961 caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct ttgccttggt
    10021 gggtgctact cctaatggtt caatttttac tactttatat ttatataatt cacttctcca
    10081 attgtccctc atatctcctc ctccaggtct gaagatcagc ggccggccgc ttgctgtgcg
    10141 gtggtcttac ttttgttttg ctcttcctct atcttgtcta aagcttcctt ggtgtctttt
    10201 atctctatcc tttgatgcac acaatagagg gttgctactg tattatataa tgatctaagt
    10261 tcttctgatc ctgtctgaag ggatggttgt agctgtccca gtatttgtct acagccttct
    10321 gatgtttcta acaggccagg attaactgcg aatcgttcta gctccctgct tgcccatact
    10381 atatgtttta atttatattt tttctttccc cctggcctta accgaatttt ttcccatcgc
    10441 gatctaattc tcccccgctt aatactgacg ctctcgcacc catctctctc cttctagcct
    10501 ccgctagtca aaatttttgg cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc
    10561 gcttcagcaa gccgagtcct gcgtcgagag agctcctctg gtttcccttt cgctttcaag
    10621 tccctgttcg ggcgccactg ctagagattt tccacactga ctaaaagggt ctgagggatc
    10681 tctagttacc agagtcacac aacagacggg cacacactac ttgaagcact caaggcaagc
    10741 tttattgagg cttaagcagt gggttcccta gttagccaga gagctcccag gctcagatct
    10801 ggtctaacca gagagaccca gtacaggcaa aaagcagctg cttatatgca ggatctgagg
    10861 gctcgccact ccccagtccc gcccaggcca cgcctccctg gaaagtcccc agcggaaagt
    10921 cccttgtagc aagctcgata tcagcagttc ttgaagtact ccggatgcag ctctcgggcc
    10981 acgtgatgaa atgctaggcg gctgtcaaac ctccactcta acacttctct ctccgggtca
    11041 tccatcccat gcaggctcac agggtgtaac aagctggtgt tctctccttt attggcctct
    11101 tctaccttat ctggctcaac tggtactagc ttgtagcacc atccaaaggt cagtggatat
    11161 ctgacccctg gccctggtgt gtagttctgc taatcaggga agtagccttg tgtgtggtag
    11221 atccacagat caaggatatc ttgtcttctt tgggagtgaa ttagcccttc caactactaa
    11281 gtttgtagta catatttaac aaatacaatt tctttaaaat gaaaataatt cagaggaatc
    11341 acaggtttag agtaaatgaa accacaggta attggcagtg gtaatagggt atggggtggg
    11401 aagtttggga tgattttggt tagcttgagt tatccagttg atccagacat gataagatac
    11461 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
    11521 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
    11581 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
    11641 aagtaaaacc tctacaaatg tggtatggct gattatgatc tctagtcaag gcactataca
    11701 tcaaatattc cttattaacc cctttacaaa ttaaaaagct aaaggtacac aatttttgag
    11761 catagttatt aatagcagac actctatgcc tgtgtggagt aagaaaaaac agtatgttat
    11821 gattataact gttatgccta cttataaagg ttacagaata tttttccata attttcttgt
    11881 atagcagtgc agctttttcc tttgtggtgt aaatagcaaa gcaagcaaga gttctattac
    11941 taaacacagc atgactcaaa aaacttagca attctgaagg aaagtccttg gggtcttcta
    12001 cctttctctt cttttttgga ggagtagaat gttgagagtc agcagtagcc tcatcatcac
    12061 tagatggcat ttcttctgag caaaacaggt tttcctcatt aaaggcattc caccactgct
    12121 cccattcatc agttccatag gttggaatct aaaatacaca aacaattaga atcagtagtt
    12181 taacacatta tacacttaaa aattttatat ttaccttaga gctttaaatc tctgtaggta
    12241 gtttgtccaa ttatgtcaca ccacagaagt aaggttcctt cacaaagatc cggggcccac
    12301 tcataaatcc agttgccgcc acggtagcca atcaccgtat cgtataaatc atcgtcggta
    12361 cgttcggcat cgctcatcac aatacgtgcc tggacgtcga ggatttcgcg tgggtcaatg
    12421 ccgcgccaga tccacatcag acggttaatc atgcgatacc agtgagggat ggttttacca
    12481 tcaagggccg actgcacagg cggttgtgcg ccgtgattaa agcggcggac tagcgtcgag
    12541 gtttcaggat gtttaaagcg gggtttgaac agggtttcgc tcaggtttgc ctgtgtcatg
    12601 gatgcagcct ccagaatact tactggaaac tattgtaacc cgcctgaagt taaaaagaac
    12661 aacgcccggc agtgccaggc gttgaaaaga ttagcgaccg gagattggcg ggacgaatac
    12721 gacgcccata tcccacggct gttcaatcca ggtatcttgc gggatatcaa caacatagtc
    12781 atcaaccagc ggacgaccag ccggttttgc gaagatggtg acaaagtgcg cttttggata
    12841 catttcacga atcgcaaccg cagtaccacc ggtatccacc aggtcatcaa taacgatgaa
    12901 gccttcgcca tcgccttctg cgcgtttcag cactttaagc tcgcgctggt tgtcgtgatc
    12961 gtagctggaa atacaaacgg tatcgacatg acgaataccc agttcacgcg ccagtaacgc
    13021 acccggtaca gaccgccacg gcttacggca ataatgcctt tccattgttc agaaggcatc
    13081 agtcggcttg cgagtttacg tgcatggatc tgcaacatgt cccaggtgac gatgtatttt
    13141 tcgctcatgt gaagtgtccc agcctgttta tctacggctt aaaaagtgtt cgaggggaaa
    13201 ataggttgcg cgagattata gagatctggc gcactaaaaa ccagtatttc acatgagtcc
    13261 gcgtcttttt acgcactgcc tctccctgac gcgggataaa gtggtattct caaacatatc
    13321 tcgcaagcct gtcttgtgtc caagcttttt gcaaaagcct aggcctccaa aaaagcctcc
    13381 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
    13441 aattagtcag ccatggggcg
//
