LOCUS       sequence-410267-       12884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables inducible expression of low-affinity Avitag-SUMO2nc; to 
            perform bioE3; nc = non-cleavable mutation; selection with 
            puromycin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12884)
  AUTHORS   Barroso-Gomila O, Merino-Cacho L, Muratore V, Perez C, Taibi V, 
            Maspero E, Azkargorta M, Iloro I, Trulsson F, Vertegaal ACO, Mayor 
            U, Elortza F, Polo S, Barrio R, Sutherland JD
  TITLE     BioE3 identifies specific substrates of ubiquitin E3 ligases.
  JOURNAL   Nat Commun. 2023 Nov 23;14(1):7656. doi: 10.1038/s41467-023-43326-8.
  PUBMED    37996419
REFERENCE   2  (bases 1 to 12884)
  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: "Nat 
            Commun."; date: "2023-11-23"; volume: "14(1):7656. doi"; pages: " 
            10.1038/s41467-023-43326-8"
COMMENT     Sequence Label: TRIPZ-bioGEF-SUMO2nc-PURO
FEATURES             Location/Qualifiers
     source          1..12884
                     /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 "
     protein_bind    2722..2740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2793..2811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2864..2882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2900..2918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2930..2968
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3067..3076
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3136..3414
                     /codon_start=1
                     /product="human small ubiquitin-related modifier 2"
                     /label=SUMO2
                     /translation="MADEKPKEGVKTENNDHINLKVAGQDGSVVQFKIKRHTPLSKLMK
                     AYCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQPTGG"
     promoter        3635..4845
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4682..4701
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(4858..4891)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     CDS             4898..5605
                     /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"
     protein_bind    complement(5606..5639)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     misc_feature    5662..6234
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5829..5846)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6056..6075
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6237..6839
                     /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     6736..6756
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6855..7443
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6908..6928)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7326..7337)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7650..7883
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7984..8001)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7990..8214
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8260..8688
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8347..8366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8557..8578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8697..8717)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8702..9031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8882..9017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8944..8963
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    10239..10360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10409..10425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10409..10425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10422..10444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10433..10449
                     /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(10457..10463,10464..10481,10482..10487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10502..10523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10640..10657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10811..11399)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10891..10910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11570..12361,12362..12430))
                     /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     12193..12212
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12431..12535)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12583..12717
                     /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 gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc
     2761 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
     2821 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga
     2881 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt
     2941 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg caccggtgct
     3001 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc
     3061 gaattcgcca ccatgggttt gaatgacata tttgaagccc agaagatcga gggtgagttc
     3121 ggcagcggcg aattcatggc cgacgaaaag cccaaggaag gagtcaagac tgagaacaac
     3181 gatcatatta atttgaaggt ggcggggcag gatggttctg tggtgcagtt taagattaag
     3241 aggcatacac cacttagtaa actaatgaaa gcctattgtg aacgacaggg attgtcaatg
     3301 aggcagatca gattccgatt tgacgggcaa ccaatcaatg aaacagacac acctgcacag
     3361 ttggaaatgg aggatgaaga tacaattgat gtgttccaac agccaacggg aggttaagcg
     3421 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3481 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3541 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3601 taaagcacgc gtggcctccg cgccgggttt tggcgcctcc gcgccgggtt ttggcgcctc
     3661 ccgcgggcgc ccccctcctc acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc
     3721 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     3781 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     3841 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     3901 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     3961 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     4021 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     4081 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     4141 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     4201 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     4261 gtgggcggca agaacccaag gtcttgaggc cttcgctaat gcgggaaagc tcttattcgg
     4321 gtgagatggg ctggggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
     4381 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     4441 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     4501 aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     4561 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     4621 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     4681 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     4741 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagactagt
     4801 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacgctag cggatccata
     4861 acttcgtata gtatacatta tacgaagtta tgccaccatg tctaggctgg acaagagcaa
     4921 agtcataaac ggagctctgg aattactcaa tggtgtcggt atcgaaggcc tgacgacaag
     4981 gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa
     5041 gcgggccctg ctcgatgccc tgccaatcga gatgctggac aggcatcata cccacttctg
     5101 ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cataccgctg
     5161 tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa
     5221 acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga
     5281 gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga
     5341 acaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc
     5401 acttctgaga caagcaattg agctgttcga ccggcaggga gccgaacctg ccttcctttt
     5461 cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc
     5521 gaccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgattt
     5581 tgaccttgac atgctccccg ggtaaataac ttcgtatagt atacattata cgaagttatg
     5641 gatccgcggc cgcaaattcc gcccctctcc ctcccccccc cctaacgtta ctggccgaag
     5701 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     5761 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     5821 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     5881 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     5941 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     6001 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     6061 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     6121 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     6181 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat aataccatgg
     6241 ccaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     6301 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     6361 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     6421 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     6481 gcgtcgaagc gggggcggtg ttcgccgaga tcggctcgcg catggccgag ttgagcggtt
     6541 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     6601 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     6661 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc tggggtgccc gccttcctgg
     6721 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     6781 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
     6841 ttcgcgtctg gaacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     6901 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     6961 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     7021 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     7081 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     7141 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     7201 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     7261 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     7321 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     7381 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     7441 cgcctggaat taattctgca gtcgagacct agaaaaacat ggagcaatca caagtagcaa
     7501 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     7561 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     7621 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
     7681 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7741 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7801 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7861 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     7921 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgtttaa acccgctgat
     7981 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     8041 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     8101 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     8161 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     8221 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     8281 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     8341 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     8401 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8461 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8521 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     8581 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     8641 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     8701 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     8761 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     8821 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     8881 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     8941 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     9001 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     9061 cggatctgat cagcacgtga tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     9121 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     9181 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     9241 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat
     9301 tccggaagtg cttgacattg gggaattaat tcagcgagag cctgacctat tgcatctccc
     9361 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     9421 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     9481 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     9541 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     9601 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     9661 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     9721 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     9781 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     9841 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     9901 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     9961 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
    10021 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
    10081 ccagcactcg tccgagggca aaggaatagc acgtgctacg agatttcgat tccaccgccg
    10141 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
    10201 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
    10261 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
    10321 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
    10381 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
    10441 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
    10501 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
    10561 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
    10621 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
    10681 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
    10741 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
    10801 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
    10861 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
    10921 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
    10981 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
    11041 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
    11101 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
    11161 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
    11221 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
    11281 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
    11341 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
    11401 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
    11461 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
    11521 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
    11581 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
    11641 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
    11701 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
    11761 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
    11821 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
    11881 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
    11941 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
    12001 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
    12061 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
    12121 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
    12181 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
    12241 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
    12301 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
    12361 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
    12421 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
    12481 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    12541 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tcaacttgtt tattgcagct
    12601 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
    12661 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac
    12721 tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact
    12781 gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt attttcattt
    12841 taaagaaatt gtatttgtta aatatgtact acaaacttag tagt
//
