LOCUS       sequence-228953-       11705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses control HA-birA* in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11705)
  AUTHORS   Kovalski JR, Bhaduri A, Zehnder AM, Neela PH, Che Y, Wozniak GG, 
            Khavari PA
  TITLE     The Functional Proximal Proteome of Oncogenic Ras Includes mTORC2.
  JOURNAL   Mol Cell. 2019 Jan 4. pii: S1097-2765(18)31031-1. doi: 
            10.1016/j.molcel.2018.12.001.
  PUBMED    30639242
REFERENCE   2  (bases 1 to 11705)
  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. 
            2019 Jan 4. pii: S1097-2765(18)31031-1. doi: 
            10.1016/j.molcel.2018.12.001."
COMMENT     Sequence Label: pLEX-uORF-HA-birA*-STOP-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..11705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     regulatory      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     enhancer        2791..3094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3095..3298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3249..3269
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3295..3319
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3411..3437
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(3438..3785,3786..3788,3789..4397)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     misc_feature    4496..5068
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4663..4680)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4890..4909
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5071..5673
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5570..5590
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5680..6268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5733..5753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6151..6162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6475..6708
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6809..6826)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6815..7039
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7085..7513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7172..7191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7382..7403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7522..7542)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7527..7856
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7707..7842
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7769..7788
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7905..8930
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7905..8930
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    9060..9181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9230..9246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9230..9246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9243..9265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9254..9270
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(9278..9284,9285..9302,9303..9308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9323..9344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9461..9478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9632..10220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9712..9731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10391..11182,11183..11251))
                     /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     11014..11033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11252..11356)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    11404..11538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
     2761 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2821 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2881 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2941 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     3001 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     3061 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     3121 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     3181 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     3241 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     3301 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     3361 acaccgactc tactagagga tctaccatgg cataacgact aatatttatg tacccatacg
     3421 atgtgcctga ctatgctaag gacaacaccg tgcccctgaa gctgatcgcc ctgctggcca
     3481 acggcgagtt ccactctggc gagcagctgg gagagaccct gggaatgagc agagccgcca
     3541 tcaacaagca catccagaca ctgagagact ggggagtgga cgtgttcacc gtgcctggca
     3601 agggctacag cctgcctgag cctatccagc tgctgaacgc caagcagatc ctgggacagc
     3661 tggatggcgg aagcgtggcc gtgctgcctg tgatcgactc caccaatcag tacctgctgg
     3721 acagaatcgg agagctgaag tccggcgacg cctgcatcgc cgagtaccag caggctggca
     3781 gaggaggcag aggacggaag tggttcagcc cattcggagc caacctgtac ctgtccatgt
     3841 tctggagact ggagcaggga cctgctgctg ccatcggact gagtctggtg atcggaatcg
     3901 tgatggccga ggtgctgaga aagctgggag ccgacaaggt gagagtgaag tggcctaatg
     3961 acctgtacct ccaggaccgc aagctggctg gcatcctggt ggagctgaca ggcaagacag
     4021 gcgatgccgc tcagatcgtg atcggagccg gaatcaacat ggccatgaga agagtggagg
     4081 agagcgtggt gaaccagggc tggatcaccc tgcaggaggc tggcatcaac ctggaccgga
     4141 acaccctggc cgccatgctg atcagagagc tgagagccgc tctggagctg ttcgagcagg
     4201 agggactggc tccttacctg agcagatggg agaagctgga caacttcatc aacagacctg
     4261 tgaagctgat catcggcgac aaggaaatct tcggcatctc cagaggaatc gacaagcagg
     4321 gagctctgct gctggagcag gacggaatca tcaagccctg gatgggcgga gaaatctccc
     4381 tgagaagcgc agagaagtga ggatccacta gtccagtgtg gtggaattct gcagatatcc
     4441 agcacagtgg cggccgctcg agtctagagg gcccgacgcg tctggccacc ggtcgcccct
     4501 ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     4561 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     4621 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     4681 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     4741 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     4801 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     4861 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     4921 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     4981 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     5041 tggttttcct ttgaaaaaca cgataatacc atggccaccg agtacaagcc cacggtgcgc
     5101 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     5161 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     5221 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     5281 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     5341 gagatcggct cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5401 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5461 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5521 gcggccgagc gcgctggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5581 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5641 acctggtgca tgacccgcaa gcccggtgcc tgagttaaca atcaacctct ggattacaaa
     5701 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     5761 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     5821 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     5881 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     5941 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     6001 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     6061 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     6121 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     6181 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     6241 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa
     6301 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     6361 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     6421 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag
     6481 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     6541 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6601 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6661 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6721 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     6781 cttgacattg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     6841 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6901 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     6961 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     7021 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     7081 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     7141 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     7201 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     7261 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     7321 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7381 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7441 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7501 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     7561 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     7621 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     7681 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     7741 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     7801 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     7861 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgatgaaa aagcctgaac
     7921 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
     7981 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
     8041 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
     8101 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga
     8161 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa
     8221 ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg
     8281 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
     8341 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
     8401 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
     8461 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
     8521 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
     8581 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
     8641 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
     8701 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
     8761 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
     8821 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
     8881 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtgctac
     8941 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     9001 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     9061 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     9121 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     9181 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     9241 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     9301 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     9361 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     9421 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     9481 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     9541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     9601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     9661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     9721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     9781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     9841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     9901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     9961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    10021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    10081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    10141 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    10201 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    10261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    10321 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    10381 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    10441 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    10501 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    10561 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    10621 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    10681 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    10741 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    10801 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    10861 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    10921 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    10981 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    11041 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    11101 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    11161 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    11221 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    11281 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    11341 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    11401 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
    11461 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    11521 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
    11581 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
    11641 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
    11701 gtagt
//
