LOCUS       sequence-385723-       10274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TGFbeta reporter driven nuclear mCherry and CreERT2 with PGK driven 
            rtTA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10274)
  AUTHORS   Oshimori N, Oristian D, Fuchs E
  TITLE     TGF-beta promotes heterogeneity and drug resistance in squamous cell
            carcinoma.
  JOURNAL   Cell. 2015 Feb 26;160(5):963-976. doi: 10.1016/j.cell.2015.01.043.
  PUBMED    25723170
REFERENCE   2  (bases 1 to 10274)
  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: 
            "10.1016/j.cell.2015.01"; journalName: "Cell"; date: "2015-02-26- 
            26"; volume: "160"; issue: "5"; pages: "963-976"
COMMENT     Sequence Label: SBE-NLS-mCherry-P2A-CreERT2 PGK-rtTA3
FEATURES             Location/Qualifiers
     source          1..10274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     426..444
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             529..1233
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     CDS             complement(1253..2146)
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     CDS             complement(1259..2194)
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             complement(2207..3235)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             complement(3236..3292)
                     /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             complement(3317..4024)
                     /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(3592..3611)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3861..3879
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(4034..4054)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     LTR             4447..4680
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4752..4873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4789..4808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4843..4862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4913..5048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5068..5087)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5069..5087)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5097..5114)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5097..5113)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5106..5128)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5255..5710
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5342..5361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5552..5573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5736..5840
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5841..5909,5910..6701)
                     /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(6059..6078)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6872..7460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7361..7380
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7614..7631
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7748..7769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7784..7789,7790..7807,7808..7814)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7822..7838
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7827..7849
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7846..7862
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7846..7862
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7881..7901
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7883..7901
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7929..8155
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8156..8336
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8383..8508
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9001..9234
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9419..9463
                     /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"
     promoter        9761..10009
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9761..9781
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9932..9951
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    10105..10222
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(10271..10274,1..507)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccaccgg agcttaccat gtctaggctg
      541 gacaagagca aagtcataaa cggagctctg gaattactca atggtgtcgg tatcgaaggc
      601 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
      661 gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat
      721 acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
      781 tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
      841 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
      901 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
      961 gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
     1021 tctatgcccc cacttctgag acaagcaatt gagctgttcg accggcaggg agccgaacct
     1081 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
     1141 agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     1201 cttgacgatt ttgaccttga catgctcccc ggggaaatgc atgactgcta gactaagctg
     1261 tggcagggaa accctctgcc tcccccgtga tgtaatactt ttgcaaggaa tgcgatgaag
     1321 tagagcccgc agtggccaag tggctttggt ccgtctcctc cacggatgcc cctccacggc
     1381 tagtgggcgc atgtaggcgg tgggcgtccg ccgcctccag cagcaggtca tagaggggca
     1441 ccacgttctt gcacttcatg ctgtacagat gctccatgcc tttgttactc atgtgcctga
     1501 tgtgggagag gatgaggagg agctgggcca gccgctggtg ctgctgctgc agggtcaggc
     1561 ctgccttggc catcaggtgg atcaaagtgt ctgtgatctt gtccaggact cggtggatat
     1621 ggtccttctc ttccagagac ttcagggtgc tggacagaaa tgtgtacact ccagaattaa
     1681 gcaaaataat agatttgagg cacacaaact cctctccctg cagattcatc atgcggaacc
     1741 gagatgatgt agccagcagc atgtcgaaga tctccaccat gccctctaca cattttccct
     1801 ggttcctgtc caagagcaag ttaggagcaa acagtagctt cactgggtgc tccatggagc
     1861 gccagacgag accaatcatc aggatctcta gccaggcaca ttcaagaagg tggacctgat
     1921 catggagggt caaatccaca aagcctggca ccctcttcgc ccagttgatc atgtgaacca
     1981 gctccctgtc tgccaggttg gtcagtaagc ccatcatcga agcttcactg aagggtctgg
     2041 taggatcata ctcggaatag agtatggggg gctcagcatc caacaaggca ctgaccatct
     2101 ggtcggccgt cagggacaag gccaggctgt tcttcttaga gcgtttgatc atgagcgggc
     2161 ttggccaaag gttggcagct ctcatgtctc cagcagatgg ctcgagatcg ccatcttcca
     2221 gcaggcgcac cattgcccct gtttcactat ccaggttacg gatatagttc atgacaatat
     2281 ttacattggt ccagccacca gcttgcatga tctccggtat tgaaactcca gcgcgggcca
     2341 tatctcgcgc ggctccgaca cgggcactgt gtccagacca ggccaggtat ctctgaccag
     2401 agtcatcctt agcgccgtaa atcaatcgat gagttgcttc aaaaatccct tccagggcgc
     2461 gagttgatag ctggctggtg gcagatggcg cggcaacacc attttttctg acccggcaaa
     2521 acaggtagtt attcggatca tcagctacac cagagacgga aatccatcgc tcgaccagtt
     2581 tagttacccc caggctaagt gccttctcta cacctgcggt gctaaccagc gttttcgttc
     2641 tgccaatatg gattaacatt ctcccaccgt cagtacgtga gatatcttta accctgatcc
     2701 tggcaatttc ggctatacgt aacagggtgt tataagcaat ccccagaaat gccagattac
     2761 gtatatcctg gcagcgatcg ctattttcca tgagtgaacg aacctggtcg aaatcagtgc
     2821 gttcgaacgc tagagcctgt tttgcacgtt caccggcatc aacgttttct tttcggatcc
     2881 gccgcataac cagtgaaaca gcattgctgt cacttggtcg tggcagcccg gaccgacgat
     2941 gaagcatgtt tagctggccc aaatgttgct ggatagtttt tactgccaga ccgcgcgcct
     3001 gaagatatag aagataatcg cgaacatctt caggttctgc gggaaaccat ttccggttat
     3061 tcaacttgca ccatgccgcc cacgaccggc aaacggacag aagcattttc caggtatgct
     3121 cagaaaacgc ctggcgatcc ctgaacatgt ccatcaggtt cttgcgaacc tcatcactcg
     3181 ttgcatcgac cggtaatgca ggcaaatttt ggtgtacggt cagtaaattg gacatagggc
     3241 cgggattctc ctccacgtca cctgcttgtt tgagtagtga gaagtttgtt gctccagatc
     3301 ctgctagctc ggccgccttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg
     3361 cgcgttcgta ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga
     3421 tgttgacgtt gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga
     3481 cctcagcgtc gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca
     3541 gggcgccgtc ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct
     3601 gcattacggg gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact
     3661 cgccgtcctg cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca
     3721 tcacgcgctc ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg
     3781 cggggtgctt cacgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg
     3841 cgaagggcag ggggccaccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg
     3901 ggcggccctc gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgt
     3961 gcaccttgaa gcgcatgaac tccttgatga tggccatgtt atcctcctcg cccttgctca
     4021 ccatcgaagc ctcaaccttt ctcttcttct ttggcgacat ggtggcgacc ggcagatctg
     4081 atcggtcccg gtgtcttcta tggaggtcaa aacagcgtgg atggcgtctc cagggatctg
     4141 acggttcact aaaccagctc tgcttatata ggcctcccac cgtacacgcc tagacttgtc
     4201 tggctcagta tgtctggctc cttctgtctg gctttcaatg tctggctaca aatgtctggc
     4261 taacgttgtc tggcttggac tgtctggctc gttatgtctg gctgaatgtg tctggctctg
     4321 attgtctggc tgctgatgtc tggcttaccc tgtctggctg actacgcgtg tcgacggtac
     4381 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     4441 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     4501 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     4561 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     4621 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     4681 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     4741 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4801 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4861 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     4921 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4981 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5041 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     5101 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     5161 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     5221 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     5281 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     5341 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     5401 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     5461 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     5521 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5581 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5641 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     5701 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5761 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5821 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     5881 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5941 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6001 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6061 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6121 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6181 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6241 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6301 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6361 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6421 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6481 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6541 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6601 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6661 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6721 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6781 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6841 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6901 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6961 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7021 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7081 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7141 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7201 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7261 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7321 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7381 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7441 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7501 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7561 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7621 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     7681 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     7741 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     7801 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     7861 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
     7921 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
     7981 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
     8041 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
     8101 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
     8161 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     8221 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     8281 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     8341 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     8401 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     8461 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     8521 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     8581 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     8641 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     8701 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     8761 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     8821 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     8881 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     8941 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     9001 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     9061 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     9121 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     9181 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     9241 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     9301 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     9361 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     9421 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     9481 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     9541 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     9601 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     9661 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     9721 tcgatcacga gactagcctc gagcggccgc ccccttcacc gagggcctat ttcccatgat
     9781 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
     9841 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     9901 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     9961 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg gcaacaagat
    10021 gaagagcacc aactcgagtt ggtgctcttc atcttgttgt ttttaattct cgacctcgag
    10081 acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc
    10141 aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca
    10201 aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcc
    10261 gcggctcgag gggg
//
