LOCUS       sequence-410266-       12833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables inducible expression of low-affinity Avitag-UBnc; 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 12833)
  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 12833)
  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-UBnc-PURO
FEATURES             Location/Qualifiers
     source          1..12833
                     /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    2865..2883
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2901..2919
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2931..2969
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3068..3077
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3137..3364
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRPRGG"
     promoter        3584..4794
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4631..4650
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(4807..4840)
                     /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             4847..5554
                     /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(5555..5588)
                     /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    5611..6183
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5778..5795)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6005..6024
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6186..6788
                     /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     6685..6705
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6804..7392
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6857..6877)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7275..7286)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7599..7832
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7933..7950)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7939..8163
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8209..8637
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8296..8315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8506..8527
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8646..8666)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8651..8980
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8831..8966
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8893..8912
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    10188..10309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10358..10374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10358..10374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10371..10393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10382..10398
                     /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(10406..10412,10413..10430,10431..10436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10451..10472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10589..10606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10760..11348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10840..10859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11519..12310,12311..12379))
                     /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     12142..12161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12380..12484)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12532..12666
                     /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 ttactcccta tcagtgatag
     2881 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg
     2941 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcaccggtgc
     3001 tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga gctcaagctt
     3061 cgaattcgcc accatgggtt tgaatgacat atttgaagcc cagaagatcg agggtgagtt
     3121 cggatccggc tccggaatgc aaatcttcgt gaagaccctg actggtaaga ccatcactct
     3181 cgaagtggag ccgagtgaca ccattgagaa tgtcaaggca aagatccaag acaaggaagg
     3241 catccctcct gaccagcaga ggttgatctt tgctgggaaa cagctggaag atggacgcac
     3301 cctgtctgac tacaacatcc agaaagagtc caccctgcac ctggtcctcc gtcctagagg
     3361 tggttaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     3421 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3481 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3541 attcacaaat aaagcacgcg tggcctccgc gccgggtttt ggcgcctccg cgccgggttt
     3601 tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc
     3661 gcagcgagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag
     3721 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact
     3781 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg
     3841 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc
     3901 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg
     3961 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc
     4021 cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg
     4081 tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt
     4141 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
     4201 gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg cgggaaagct
     4261 cttattcggg tgagatgggc tggggcacca tctggggacc ctgacgtgaa gtttgtcact
     4321 gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg
     4381 cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg
     4441 ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt
     4501 cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc
     4561 tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt
     4621 cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt
     4681 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt
     4741 tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgctagc
     4801 ggatccataa cttcgtatag tatacattat acgaagttat gccaccatgt ctaggctgga
     4861 caagagcaaa gtcataaacg gagctctgga attactcaat ggtgtcggta tcgaaggcct
     4921 gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt
     4981 gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac
     5041 ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc
     5101 ataccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc
     5161 aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt
     5221 ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt
     5281 attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc
     5341 tatgccccca cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc
     5401 cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag
     5461 cggcgggccg accgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct
     5521 tgacgatttt gaccttgaca tgctccccgg gtaaataact tcgtatagta tacattatac
     5581 gaagttatgg atccgcggcc gcaaattccg cccctctccc tccccccccc ctaacgttac
     5641 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     5701 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     5761 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     5821 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     5881 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     5941 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     6001 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     6061 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     6121 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata
     6181 ataccatggc caccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc
     6241 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     6301 acccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     6361 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     6421 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggctcgcgc atggccgagt
     6481 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     6541 ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg
     6601 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgct ggggtgcccg
     6661 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     6721 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     6781 gtgcctgagt tcgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga
     6841 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     6901 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     6961 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     7021 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     7081 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     7141 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     7201 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     7261 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     7321 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     7381 ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac
     7441 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     7501 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     7561 tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa ttcactccca
     7621 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     7681 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     7741 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     7801 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     7861 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac attgtttaaa
     7921 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     7981 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     8041 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     8101 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     8161 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     8221 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     8281 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     8341 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     8401 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     8461 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     8521 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     8581 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     8641 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     8701 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     8761 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     8821 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     8881 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     8941 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     9001 atccattttc ggatctgatc agcacgtgat gaaaaagcct gaactcaccg cgacgtctgt
     9061 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     9121 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     9181 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     9241 gctcccgatt ccggaagtgc ttgacattgg ggaattaatt cagcgagagc ctgacctatt
     9301 gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg
     9361 ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga
     9421 cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt
     9481 tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
     9541 tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg
     9601 aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc
     9661 gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg
     9721 ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg
     9781 agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg
     9841 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc
     9901 agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg
     9961 cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa
    10021 accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtgctacga gatttcgatt
    10081 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
    10141 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
    10201 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    10261 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
    10321 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    10381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    10441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    10501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    10561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    10621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    10681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    10741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    10801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    10861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    10921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    10981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    11041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    11101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    11161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
    11221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    11281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    11341 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
    11401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
    11461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
    11521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
    11581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
    11641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
    11701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
    11761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    11821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    11881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    11941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    12001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    12061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    12121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    12181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    12241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    12301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    12361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    12421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    12481 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt
    12541 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
    12601 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    12661 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct
    12721 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta
    12781 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
