LOCUS       sequence-402526-       10309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-ON 3rd generation expression vector with inducible expression of
            PSMD4 and a puromycin selection cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10309)
  AUTHORS   Girish V, Lakhani AA, Thompson SL, Scaduto CM, Brown LM, Hagenson 
            RA, Sausville EL, Mendelson BE, Kandikuppa PK, Lukow DA, Yuan ML, 
            Stevens EC, Lee SN, Schukken KM, Akalu SM, Vasudevan A, Zou C, 
            Salovska B, Li W, Smith JC, Taylor AM, Martienssen RA, Liu Y, Sun R,
            Sheltzer JM
  TITLE     Oncogene-like addiction to aneuploidy in human cancers.
  JOURNAL   Science. 2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521.
  PUBMED    37410869
REFERENCE   2  (bases 1 to 10309)
  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: "Science. 
            2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521."
COMMENT     Sequence Label: pLVX-TetOne-Puro-PSMD4
FEATURES             Location/Qualifiers
     source          1..10309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..377
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    18..36
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    54..72
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    90..108
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    126..144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    162..180
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    198..216
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    234..252
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     polyA_signal    1713..1847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    1891..2006
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(2269..2313)
                     /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    2498..2731
                     /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    3228..3353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             3400..4033
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    4201..4335
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4383..4487
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4488..4556,4557..5348)
                     /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(4706..4725)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5519..6107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6008..6027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6261..6278
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6395..6416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6431..6436,6437..6454,6455..6461)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6469..6485
                     /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     6474..6496
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6493..6509
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6493..6509
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             6637..7270
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7477..8065
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7583..7594
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7992..8012
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8079..8678)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8162..8182)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     8659..8678
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(8687..9016)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8701..8836
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             complement(9027..9773)
                     /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"
     promoter        complement(9792..10302)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9855..9873)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
ORIGIN
        1 tatcacctcg agtttactcc ctatcagtga tagagaacgt atgaagagtt tactccctat
       61 cagtgataga gaacgtatgc agactttact ccctatcagt gatagagaac gtataaggag
      121 tttactccct atcagtgata gagaacgtat gaccagttta ctccctatca gtgatagaga
      181 acgtatctac agtttactcc ctatcagtga tagagaacgt atatccagtt tactccctat
      241 cagtgataga gaacgtataa gctttgctta tgtaaaccag ggcgcctata aaagagtgct
      301 gattttttga gtaaacttca attccacaac acttttgtct tataccaact ttccgtacca
      361 cttcctaccc tcgtaaagaa ttcggcatgg tgttggaaag cactatggtg tgtgtggaca
      421 acagtgagta tatgcggaat ggagacttct tacccaccag gctgcaggcc cagcaggatg
      481 ctgtcaacat agtttgtcat tcaaagaccc gcagcaaccc tgagaacaac gtgggcctta
      541 tcacactggc taatgactgt gaagtgctga ccacactcac cccagacact ggccgtatcc
      601 tgtccaagct acatactgtc caacccaagg gcaagatcac cttctgcacg ggcatccgcg
      661 tggcccatct ggctctgaag caccgacaag gcaagaatca caagatgcgc atcattgcct
      721 ttgtgggaag cccagtggag gacaatgaga aggatctggt gaaactggct aaacgcctca
      781 agaaggagaa agtaaatgtt gacattatca attttgggga agaggaggtg aacacagaaa
      841 agctgacagc ctttgtaaac acgttgaatg gcaaagatgg aaccggttct catctggtga
      901 cagtgcctcc tgggcccagt ttggctgatg ctctcatcag ttctccgatt ttggctggtg
      961 aaggtggtgc catgctgggt cttggtgcca gtgactttga atttggagta gatcccagtg
     1021 ctgatcctga gctggccttg gcccttcgtg tatctatgga agagcagcgg cagcggcagg
     1081 aggaggaggc ccggcgggca gctgcagctt ctgctgctga ggccgggatt gctacgactg
     1141 ggactgaaga ctcagacgat gccctgctga agatgaccat cagccagcaa gagtttggcc
     1201 gcactgggct tcctgaccta agcagtatga ctgaggaaga gcagattgct tatgccatgc
     1261 agatgtccct gcagggagca gagtttggcc aggcggaatc agcagacatt gatgccagct
     1321 cagctatgga cacatctgag ccagccaagg aggaggatga ttacgacgtg atgcaggacc
     1381 ccgagttcct tcagagtgtc ctagagaacc tcccaggtgt ggatcccaac aatgaagcca
     1441 ttcgaaatgc tatgggctcc ctggcctccc aggccaccaa ggacggcaag aaggacaaga
     1501 aggaggaaga caagaagtaa accggtgccg gcggatccag accacctccc ctgcgagcta
     1561 agctggacag ccaatgacgg gtaagagagt gacatttttc actaacctaa gacaggaggg
     1621 ccgtcagagc tactgcctaa tccaaagacg ggtaaaagtg ataaaaatgt atcactccaa
     1681 cctaagacag gcgcagcttc cgagggattt gagatccaga catgataaga tacattgatg
     1741 agtttggaca aaccaaaact agaatgcagt gaaaaaaatg ccttatttgt gaaatttgtg
     1801 atgctattgc cttatttgta accattataa gctgcaataa acaagttaag tcgataaact
     1861 ggatctctgc tgtccctgta ataaacccga aaattttgaa tttttgtaat ttgtttttgt
     1921 aattctttag tttgtatgtc tgttgctatt atgtctacta ttctttcccc tgcactgtac
     1981 cccccaatcc ccccttttct tttaaaggcg ataccgtcga gatccgttca ctaatcgaat
     2041 ggatctgtct ctgtctctct ctccaccttc ttcttctatt ccttcgggcc tgtcgggtcc
     2101 cctcggggtt gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt
     2161 cactatagaa agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat
     2221 gaataatttt atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca
     2281 tttatctaat tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat
     2341 taaggagtgt attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt
     2401 gtgattccaa atctgttcca gagatttatt actccaacta gcattccaag gcacagcagt
     2461 ggtgcaaatg agttttccag agcaacccca aatccccagg agctgttgat cctttaggta
     2521 tctttccaca gccaggattc ttgcctggag ctgcttgatg ccccagactg tgagttgcaa
     2581 cagatgctgt tgcgcctcaa tagccctcag caaattgttc tgctgctgca ctataccaga
     2641 caataattgt ctggcctgta ccgtcagcgt cattgacgct gcgcccatag tgcttcctgc
     2701 tgctcccaag aacccaagga acaaagctcc tattcccact gctctttttt ctctctgcac
     2761 cactcttctc tttgccttgg tgggtgctac tcctaatggt tcaattttta ctactttata
     2821 tttatataat tcacttctcc aattgtccct catatctcct cctccaggtc tgaagatcag
     2881 cggccggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct
     2941 aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact
     3001 gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc
     3061 agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct
     3121 agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt
     3181 aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac
     3241 ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc
     3301 ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct
     3361 ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg
     3421 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta
     3481 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag
     3541 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca aaaagcagct
     3601 gcttatatgc aggatctgag ggctcgccac tccccagtcc cgcccaggcc acgcctccct
     3661 ggaaagtccc cagcggaaag tcccttgtag caagctcgat atcagcagtt cttgaagtac
     3721 tccggatgca gctctcgggc cacgtgatga aatgctaggc ggctgtcaaa cctccactct
     3781 aacacttctc tctccgggtc atccatccca tgcaggctca cagggtgtaa caagctggtg
     3841 ttctctcctt tattggcctc ttctacctta tctggctcaa ctggtactag cttgtagcac
     3901 catccaaagg tcagtggata tctgacccct ggccctggtg tgtagttctg ctaatcaggg
     3961 aagtagcctt gtgtgtggta gatccacaga tcaaggatat cttgtcttct ttgggagtga
     4021 attagccctt ccaactacta agtttgtagt acatatttaa caaatacaat ttctttaaaa
     4081 tgaaaataat tcagaggaat cacaggttta gagtaaatga aaccacaggt aattggcagt
     4141 ggtaataggg tatggggtgg gaagtttggg atgattttgg ttagcttgag ttatccagtt
     4201 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     4261 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     4321 tgcaataaac aagttgatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
     4381 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4441 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4501 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4561 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4621 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4681 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4741 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4801 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4861 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4921 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4981 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5041 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5101 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5161 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5221 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5281 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5341 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5401 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5461 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5521 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5581 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5641 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5701 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5761 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5821 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5881 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5941 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6001 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6061 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6121 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6181 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6241 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6301 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6361 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     6421 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     6481 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     6541 acggtaaacg ctagcaatgt caaggcctct cactctctga tattcatttc tttgcaagtt
     6601 ataaatactg aataataaga tgacatgaac tactactgct agagattttc cacactgact
     6661 aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt
     6721 gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga
     6781 gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa agcagctgct
     6841 tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg cctccctgga
     6901 aagtccccag cggaaagtcc cttgtagcaa gctcgatatc agcagttctt gaagtactcc
     6961 ggatgcagct ctcgggccac gtgatgaaat gctaggcggc tgtcaaacct ccactctaac
     7021 acttctctct ccgggtcatc catcccatgc aggctcacag ggtgtaacaa gctggtgttc
     7081 tctcctttat tggcctcttc taccttatct ggctcaactg gtactagctt gtagcaccat
     7141 ccaaaggtca gtggatatct gacccctggc cctggtgtgt agttctgcta atcagggaag
     7201 tagccttgtg tgtggtagat ccacagatca aggatatctt gtcttcgttg ggagtgaatt
     7261 agcccttcca gtcccctctt ttcttttaaa aagtggctaa gatctacagc tgccttgtaa
     7321 gtcattggtc ttaaaggtac ctgaggtgtg actggaaaac ccacctcctc ctcctcttgt
     7381 gcttctagcc aggcacaatc agcattggta gctgctgtat tgctacttgt gattgctcca
     7441 tgtttttcta ggtctcgact gcagaattaa ttccaggcgg ggaggcggcc caaagggaga
     7501 tccgactcgt ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc
     7561 cgcgggaagg aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc
     7621 gcagaatcca ggtggcaaca caggcgagca gccatggaaa ggacgtcagc ttccccgaca
     7681 acaccacgga attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg
     7741 cggcgatgag ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct
     7801 gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca
     7861 cgccacgttg cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat
     7921 acaaggagga gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag
     7981 cagcgtatcc acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac
     8041 aaattttgta atccagaggt tgattgttcc agacgcgttc aggcaccggg cttgcgggtc
     8101 atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc
     8161 cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg
     8221 cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg
     8281 tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc
     8341 aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc
     8401 gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc
     8461 accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg
     8521 aagagttctt gcagctcggt gacccgctcg atgtggcggt ccgggtcgac ggtgtggcgc
     8581 gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggcccgggg gacgtcgtcg
     8641 cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg tcgcgttttg caaaagccta
     8701 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     8761 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     8821 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     8881 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     8941 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     9001 accctaactg acacacattc gcgcgtttac ccggggagca tgtcaaggtc aaaatcgtca
     9061 agagcgtcag caggcagcat atcaaggtca aagtcgtcaa gggcatcggc tgggagcatg
     9121 tctaagtcaa aatcgtcaag ggcgtcggtc ggcccgccgc tttcgcactt tagctgtttc
     9181 tccaggccac atatgattag ttccaggccg aaaaggaagg caggttcggc tccctgccgg
     9241 tcgaacagct caattgcttg tttcagaagt gggggcatag aatcggtggt aggtgtctct
     9301 ctttcctctt ttgctacttg atgctcctgt tcctccaata cgcagcccag tgtaaagtgg
     9361 cccacggcgg acagagcgta cagtgcgttc tccagggaga agccttgctg acacaggaac
     9421 gcgagctgat tttccagggt ttcgtactgt ttctctgttg ggcgggtgcc gagatgcact
     9481 ttagccccgt cgcgatgtga gaggagagca cagcggtatg acttggcgtt gttccgcaga
     9541 aagtcttgcc atgactcgcc ttccaggggg caggagtggg tatgatgcct gtccagcatc
     9601 tcgattggca gggcatcgag cagggcccgc ttgttcttca cgtgccagta cagggtaggc
     9661 tgctcaactc ccagcttttg agcgagtttc cttgtcgtca ggccttcgat accgactcca
     9721 ttgagtaatt ccagagcaga gtttatgact ttgctcttgt ccagtctaga catggtaatt
     9781 cgatgatccc cctggggaga gaggtcggtg attcggtcaa cgagggagcc gactgccgac
     9841 gtgcgctccg gaggcttgca gaatgcggaa caccgcgcgg gcaggaacag ggcccacact
     9901 accgccccac accccgcctc ccgcaccgcc ccttcccggc cgctgctctc ggcgcgccct
     9961 gctgagcagc cgctattggc cacagcccat cgcggtcggc gcgctgccat tgctccctgg
    10021 cgctgtccgt ctgcgagggt actagtgaga cgtgcggctt ccgtttgtca cgtccggcac
    10081 gccgcgaacc gcaaggaacc ttcccgactt aggggcggag caggaagcgt cgccgggggg
    10141 cccacaaggg tagcggcgaa gatccgggtg acgctgcgaa cggacgtgaa gaatgtgcga
    10201 gacccagggt cggcgccgct gcgtttcccg gaaccacgcc cagagcagcc gcgtccctgc
    10261 gcaaacccag ggctgccttg gaaaaggcgc aaccccaacc ccgtggaat
//
