LOCUS       sequence-410263-       12928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables inducible expression of low-affinity Avitag-SUMO1nc; 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 12928)
  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 12928)
  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-SUMO1nc-PURO
FEATURES             Location/Qualifiers
     source          1..12928
                     /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      3099..3108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        3679..4889
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4726..4745
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(4902..4935)
                     /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             4942..5649
                     /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(5650..5683)
                     /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    5706..6278
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5873..5890)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6100..6119
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6281..6883
                     /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     6780..6800
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6899..7487
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6952..6972)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7370..7381)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7694..7927
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8028..8045)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8034..8258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8304..8732
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8391..8410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8601..8622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8741..8761)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8746..9075
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8926..9061
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8988..9007
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    10283..10404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10453..10469)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10453..10469)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10466..10488)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10477..10493
                     /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(10501..10507,10508..10525,10526..10531))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10546..10567
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10684..10701)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10855..11443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10935..10954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11614..12405,12406..12474))
                     /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     12237..12256
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12475..12579)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12627..12761
                     /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 cgcaccggtg ctggtttagt gaaccgtcag atccgctagc
     3061 gctaccggac tcagatctcg agctcaagct tcgaattcgc caccatgggt ttgaatgaca
     3121 tatttgaagc ccagaagatc gagggtgagt tcggcagcgg cgaattcatg tctgaccagg
     3181 aggcaaaacc ttcaactgag gacttggggg ataagaagga aggtgaatat attaaactca
     3241 aagtcattgg acaggatagc agtgagattc acttcaaagt gaaaatgaca acacatctca
     3301 agaaactcaa agaatcatac tgtcaaagac agggtgttcc aatgaatagc ctcaggtttc
     3361 tctttgaggg tcagagaatt gctgataatc atactccaaa agaactggga atggaggaag
     3421 aagatgtgat tgaagtttat caggaaccta cgggtggtta agcggccgcg actctagatc
     3481 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     3541 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     3601 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc acgcgtggcc
     3661 tccgcgccgg gttttggcgc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     3721 cctcacggcg agcgctgcca cgtcagacga agggcgcagc gagcgtcctg atccttccgc
     3781 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     3841 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     3901 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     3961 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     4021 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     4081 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     4141 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     4201 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     4261 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     4321 caaggtcttg aggccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggg
     4381 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     4441 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     4501 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     4561 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     4621 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     4681 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     4741 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     4801 tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat
     4861 tctggccgtt tttggctttt ttgttagacg ctagcggatc cataacttcg tatagtatac
     4921 attatacgaa gttatgccac catgtctagg ctggacaaga gcaaagtcat aaacggagct
     4981 ctggaattac tcaatggtgt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag
     5041 ctgggagttg agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat
     5101 gccctgccaa tcgagatgct ggacaggcat catacccact tctgccccct ggaaggcgag
     5161 tcatggcaag actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat
     5221 cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg
     5281 gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct
     5341 ctgtccgccg tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta
     5401 gcaaaagagg aaagagagac acctaccacc gattctatgc ccccacttct gagacaagca
     5461 attgagctgt tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc
     5521 atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac
     5581 gattttgact tagacatgct cccagccgat gcccttgacg attttgacct tgacatgctc
     5641 cccgggtaaa taacttcgta tagtatacat tatacgaagt tatggatccg cggccgcaaa
     5701 ttccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     5761 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     5821 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     5881 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     5941 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     6001 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     6061 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     6121 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     6181 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     6241 cacggggacg tggttttcct ttgaaaaaca cgataatacc atggccaccg agtacaagcc
     6301 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
     6361 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
     6421 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     6481 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     6541 ggtgttcgcc gagatcggct cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     6601 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     6661 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     6721 cggagtggag gcggccgagc gcgctggggt gcccgccttc ctggagacct ccgcgccccg
     6781 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     6841 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgagttcgcg tctggaacaa
     6901 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     6961 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     7021 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     7081 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     7141 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     7201 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     7261 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     7321 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     7381 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     7441 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc
     7501 tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     7561 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     7621 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     7681 gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     7741 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7801 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7861 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7921 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     7981 atatcagaga gtgagaggcc ttgacattgt ttaaacccgc tgatcagcct cgactgtgcc
     8041 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     8101 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     8161 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     8221 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     8281 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     8341 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     8401 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     8461 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     8521 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     8581 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     8641 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     8701 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     8761 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     8821 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     8881 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     8941 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     9001 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     9061 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     9121 gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     9181 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     9241 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     9301 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     9361 attggggaat taattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     9421 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     9481 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     9541 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     9601 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     9661 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     9721 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     9781 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     9841 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     9901 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     9961 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
    10021 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
    10081 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
    10141 ggcaaaggaa tagcacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
    10201 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
    10261 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
    10321 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    10381 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
    10441 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
    10501 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
    10561 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
    10621 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
    10681 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
    10741 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
    10801 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
    10861 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
    10921 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
    10981 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
    11041 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
    11101 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
    11161 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
    11221 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
    11281 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
    11341 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
    11401 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
    11461 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
    11521 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
    11581 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
    11641 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
    11701 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
    11761 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
    11821 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
    11881 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
    11941 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
    12001 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
    12061 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
    12121 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
    12181 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
    12241 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
    12301 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
    12361 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
    12421 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
    12481 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
    12541 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    12601 acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat ggttacaaat
    12661 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    12721 gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata actcaagcta
    12781 accaaaatca tcccaaactt cccaccccat accctattac cactgccaat tacctgtggt
    12841 ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga aattgtattt
    12901 gttaaatatg tactacaaac ttagtagt
//
