LOCUS       sequence-376586-       12854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heat inducible Fluc reporter with constitutive Rluc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12854)
  AUTHORS   Wu Y, Liu Y, Huang Z, Wang X, Jin Z, Li J, Limsakul P, Zhu L, Allen 
            M, Pan Y, Bussell R, Jacobson A, Liu T, Chien S, Wang Y
  TITLE     Control of the activity of CAR-T cells within tumours via focused 
            ultrasound.
  JOURNAL   Nat Biomed Eng. 2021 Nov;5(11):1336-1347. doi: 
            10.1038/s41551-021-00779-w. Epub 2021 Aug 12.
  PUBMED    34385696
REFERENCE   2  (bases 1 to 12854)
  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: 
            "10.1038/s41551-021-00779-w"; journalName: "Nat Biomed Eng"; date: 
            "2021-11"; volume: "5"; issue: "11"; pages: "1336-1347"
COMMENT     Sequence Label: pHR-Hsp-Fluc-PGK-Rluc-mCherry
FEATURES             Location/Qualifiers
     source          1..12854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             23..256
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(281..299)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(282..299)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(589..608)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(822..841)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     941..963
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1001..1019)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1087..1191
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1192..1260,1261..2052)
                     /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(1410..1429)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2223..2811
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2712..2731
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2965..2982
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3052..3072)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3057..3386
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3237..3372
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          4520..4585
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4715..4735
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5160..5294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5197..5216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5251..5270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             5462..6095
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6142..6267
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6764..6997
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7182..7226
                     /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    7511..7628
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      8138..8147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8144..9796
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(8210..8228)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        9807..10306
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9842..9865)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     10239..10258
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             10327..11259
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             11269..11325
                     /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             11335..11391
                     /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             11392..12102
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(11537..11555)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     11805..11824
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    12179..12767
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12232..12252)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12650..12661)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12770..12786)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
       61 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      121 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      181 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      241 tggaaaatct ctagcagcat ctagaattaa ttccgtgtat tctatagtgt cacctaaatc
      301 gtatgtgtat gatacataag gttatgtatt aattgtagcc gcgttctaac gacaatatgt
      361 acaagcctaa ttgtgtagca tctggcttac tgaagcagac cctatcatct ctctcgtaaa
      421 ctgccgtcag agtcggtttg gttggacgaa ccttctgagt ttctggtaac gccgtcccgc
      481 acccggaaat ggtcagcgaa ccaatcagca gggtcatcgc tagccagatc ctctacgccg
      541 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
      601 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
      661 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
      721 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
      781 aggagtcgca taagggagag cgtcgaatgg tgcactctca gtacaatctg ctctgatgcc
      841 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      901 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
      961 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     1021 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     1081 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1141 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1201 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1261 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1321 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1381 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     1441 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     1501 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     1561 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     1621 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     1681 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     1741 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     1801 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     1861 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     1921 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1981 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2041 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2341 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2521 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2581 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2641 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2701 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2761 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2821 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2881 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     2941 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     3001 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg tggaatgtgt
     3061 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3121 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3181 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3241 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3301 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3361 tttttggagg cctaggcttt tgcaaaaagc ttggacacaa gacaggcttg cgagatatgt
     3421 ttgagaatac cactttatcc cgcgtcaggg agaggcagtg cgtaaaaaga cgcggactca
     3481 tgtgaaatac tggtttttag tgcgccagat ctctataatc tcgcgcaacc tattttcccc
     3541 tcgaacactt tttaagccgt agataaacag gctgggacac ttcacatgag cgaaaaatac
     3601 atcgtcacct gggacatgtt gcagatccat gcacgtaaac tcgcaagccg actgatgcct
     3661 tctgaacaat ggaaaggcat tattgccgta agccgtggcg gtctgtaccg ggtgcgttac
     3721 tggcgcgtga actgggtatt cgtcatgtcg ataccgtttg tatttccagc tacgatcacg
     3781 acaaccagcg cgagcttaaa gtgctgaaac gcgcagaagg cgatggcgaa ggcttcatcg
     3841 ttattgatga cctggtggat accggtggta ctgcggttgc gattcgtgaa atgtatccaa
     3901 aagcgcactt tgtcaccatc ttcgcaaaac cggctggtcg tccgctggtt gatgactatg
     3961 ttgttgatat cccgcaagat acctggattg aacagccgtg ggatatgggc gtcgtattcg
     4021 tcccgccaat ctccggtcgc taatcttttc aacgcctggc actgccgggc gttgttcttt
     4081 ttaacttcag gcgggttaca atagtttcca gtaagtattc tggaggctgc atccatgaca
     4141 caggcaaacc tgagcgaaac cctgttcaaa ccccgcttta aacatcctga aacctcgacg
     4201 ctagtccgcc gctttaatca cggcgcacaa ccgcctgtgc agtcggccct tgatggtaaa
     4261 accatccctc actggtatcg catgattaac cgtctgatgt ggatctggcg cggcattgac
     4321 ccacgcgaaa tcctcgacgt ccaggcacgt attgtgatga gcgatgccga acgtaccgac
     4381 gatgatttat acgatacggt gattggctac cgtggcggca actggattta tgagtgggcc
     4441 ccggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac
     4501 agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactact
     4561 gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat gggagcagtg
     4621 gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga
     4681 tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa aggtagaaga
     4741 ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt ttagtaatag
     4801 aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc tatacaagaa
     4861 aattatggaa aaatattctg taacctttat aagtaggcat aacagttata atcataacat
     4921 actgtttttt cttactccac acaggcatag agtgtctgct attaataact atgctcaaaa
     4981 attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt tgatgtatag
     5041 tgccttgact agagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     5101 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     5161 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5221 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5281 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     5341 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
     5401 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     5461 ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg gatctaccac
     5521 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
     5581 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
     5641 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
     5701 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
     5761 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
     5821 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
     5881 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
     5941 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     6001 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     6061 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
     6121 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     6181 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     6241 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     6301 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     6361 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     6421 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     6481 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     6541 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     6601 caagcggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     6661 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     6721 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     6781 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     6841 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     6901 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     6961 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     7021 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     7081 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     7141 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     7201 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     7261 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     7321 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     7381 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     7441 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgccaa atggcagtat
     7501 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     7561 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     7621 aaaattttcg ggtttattac agggacagca gagatccagt ttggatcgat aagcttgata
     7681 tcgtcgaggc gcgtcctcag agccagccgg gaggagctag aaccttcccc gcgtttcttt
     7741 cagcagccct gagtcagagg cgggctggcc tggcatagcc gcccagcctc tcggctcacg
     7801 gcccgatccg cccgaacctt ctcccggggt cagcgccgcg ctgcgccgcc cggctgactc
     7861 agcccgggcg ggcgggcggg aggctctcga ctgggcggga aggtgcggga aggttcgcgg
     7921 cggcggggtc ggggaggtgc aaaaggatga aaagcccgtg gaagcggagc tgagcagatc
     7981 cgagccgggc tggcggcaga gaaaccgcag ggagagcctc actgctgagc gcccctcgac
     8041 ggcggagcgg cagcagcctc cgtggcctcc agcatccgac aagaagctct ctagtcgacg
     8101 gtatcgataa gcttggcatt ccggtactgt tggtaaagcc accatggaag acgccaaaaa
     8161 cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact
     8221 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
     8281 tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc
     8341 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     8401 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     8461 cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt
     8521 gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat
     8581 ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac
     8641 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt
     8701 cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa
     8761 aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat
     8821 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     8881 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     8941 tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct
     9001 gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt
     9061 atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc
     9121 ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac
     9181 atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt
     9241 tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca
     9301 aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga
     9361 agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg
     9421 ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg
     9481 ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga
     9541 cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt
     9601 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     9661 aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     9721 taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg
     9781 aaagatcgcc gtgtaagaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     9841 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     9901 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     9961 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
    10021 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
    10081 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
    10141 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
    10201 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
    10261 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag
    10321 cttaccatgg cttccaaggt gtacgacccc gagcaacgca aacgcatgat cactgggcct
    10381 cagtggtggg ctcgctgcaa gcaaatgaac gtgctggact ccttcatcaa ctactatgat
    10441 tccgagaagc acgccgagaa cgccgtgatt tttctgcatg gtaacgctgc ctccagctac
    10501 ctgtggaggc acgtcgtgcc tcacatcgag cccgtggcta gatgcatcat ccctgatctg
    10561 atcggaatgg gtaagtccgg caagagcggg aatggctcat atcgcctcct ggatcactac
    10621 aagtacctca ccgcttggtt cgagctgctg aaccttccaa agaaaatcat ctttgtgggc
    10681 cacgactggg gggcttgtct ggcctttcac tactcctacg agcaccaaga caagatcaag
    10741 gccatcgtcc atgctgagag tgtcgtggac gtgatcgagt cctgggacga gtggcctgac
    10801 atcgaggagg atatcgccct gatcaagagc gaagagggcg agaaaatggt gcttgagaat
    10861 aacttcttcg tcgagaccat gctcccaagc aagatcatgc ggaaactgga gcctgaggag
    10921 ttcgctgcct acctggagcc attcaaggag aagggcgagg ttagacggcc taccctctcc
    10981 tggcctcgcg agatccctct cgttaaggga ggcaagcccg acgtcgtcca gattgtccgc
    11041 aactacaacg cctaccttcg ggccagcgac gatctgccta agatgttcat cgagtccgac
    11101 cctgggttct tttccaacgc tattgtcgag ggagctaaga agttccctaa caccgagttc
    11161 gtgaaggtga agggcctcca cttcagccag gaggacgctc cagatgaaat gggtaagtac
    11221 atcaagagct tcgtggagcg cgtgctgaag aacgagcagg gaagcggagc tactaacttc
    11281 agcctgctga agcaggctgg agacgtggag gagaaccctg gacctggttc cggcgcaaca
    11341 aacttctctc tgctgaaaca agccggagat gtcgaagaga atcctggacc gatggtgagc
    11401 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
    11461 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
    11521 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
    11581 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
    11641 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
    11701 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
    11761 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
    11821 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
    11881 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
    11941 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
    12001 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
    12061 ggccgccact ccaccggcgg catggacgag ctgtacaagt aaagcggccg cgactctaga
    12121 gtcgacctgc aggcatgcaa gcttgcaggc atgcaagctt gatatcaagc ttatcgataa
    12181 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
    12241 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
    12301 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
    12361 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
    12421 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
    12481 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
    12541 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
    12601 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
    12661 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
    12721 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
    12781 acctcgaggg aattaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg
    12841 tagatcttag ccac
//
