LOCUS       sequence-410205-       10262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enables constitutive expression of N-terminal BirAopt-fused RNF4ca 
            (catalytically inactive RING mutant); to perform bioE3; selection 
            with blasticidin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10262)
  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 10262)
  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: Lenti-EFS-BirAopt-GSQ-RNF4ca-P2A-BLAST
FEATURES             Location/Qualifiers
     source          1..10262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2654..2865
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2878..3840
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             4492..4548
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4549..4947
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    4975..5563
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5028..5048)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5446..5457)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5566..5582)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5567..5583)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6088..6268
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6294..6311)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6300..6524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6570..6998
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6657..6676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6867..6888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7007..7027)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7172..7306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7233..7252
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7368..7415
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7434..7808
                     /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"
     polyA_signal    7938..8059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7975..7994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8029..8048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8108..8124)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8108..8124)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8121..8143)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8132..8148
                     /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(8156..8162,8163..8180,8181..8186))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8201..8222
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8339..8356)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8510..9098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8590..8609)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9269..10060,10061..10129))
                     /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     9892..9911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10130..10234)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta attagctagc taggtcttga aaggagtggg aattggctcc
     2641 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     2701 gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     2761 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     2821 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggaccg gtccaccatg
     2881 aaggacaaca ccgtgcccct gaagctgatc gccctgctgg ccaacggcga gttccactct
     2941 ggcgagcagc tgggagagac cctgggaatg agcagagccg ccatcaacaa gcacatccag
     3001 acactgagag actggggagt ggacgtgttc accgtgcctg gcaagggcta cagcctgcct
     3061 gagcctatcc agctgctgaa cgccaagcag atcctgggac agctggatgg cggaagcgtg
     3121 gccgtgctgc ctgtgatcga ctccaccaat cagtacctgc tggacagaat cggagagctg
     3181 aagtccggcg acgcctgcat cgccgagtac cagcaggctg gcagaggacg cagaggacgg
     3241 aagtggttca gcccattcgg agccaacctg tacctgtcca tgttctggag actggagcag
     3301 ggacctgctg ctgccatcgg actgagtctg gtgatcggaa tcgtgatggc cgaggtgctg
     3361 agaaagctgg gagccgacaa ggtgagagtg aagtggccta atgacctgta cctccaggac
     3421 cgcaagctgg ctggcatcct ggtggagctg acaggcaaga caggcgatgc cgctcagatc
     3481 gtgatcggag ccggaatcaa catggccatg agaagagtgg aggagagcgt ggtgaaccag
     3541 ggctggatca ccctgcagga ggctggcatc aacctggacc ggaacaccct ggccgccatg
     3601 ctgatcagag agctgagagc cgctctggag ctgttcgagc aggagggact ggctccttac
     3661 ctgagcagat gggagaagct ggacaacttc atcaacagac ctgtgaagct gatcatcggc
     3721 gacaaggaaa tcttcggcat ctccagagga atcgacaagc agggagctct gctgctggag
     3781 caggacggaa tcatcaagcc ctggatgggc ggagaaatct ccctgagaag cgcagagaag
     3841 gggcgcgccg gcgggggatc ttccggaggc gggcaaattt cttacgcaag tagaggggaa
     3901 ttcatgagta caagaaagcg tcgtggtgga gcaataaatt ctagacaagc tcagaagcga
     3961 actcgggaag caacctccac ccccgagatc tccttggaag cagaacccat agaactcgtg
     4021 gaaactgctg gagatgaaat tgtggacctc acttgtgaat ctttagagcc tgtggtggtt
     4081 gatctgactc acaatgactc tgttgtgatt gttgacgaaa gaagaagacc aaggaggaat
     4141 gctaggaggc tgccccagga ccatgctgac agctgtgtgg tgagcagtga cgatgaggag
     4201 ttgtccaggg acagagacgt atatgtgact acccatactc ccagaaacgc cagggatgag
     4261 ggcgctacag gcctcaggcc ctcaggtact gtcagttgtc ccatctgcat ggacggatac
     4321 tcagagatcg tgcagaatgg acgtctcatc gtttccacag aatgcggcca tgtcttcgct
     4381 agccagtgcc tccgtgattc cctgaagaat gctaatactt gcccaacttg taggaaaaag
     4441 atcaaccaca aacggtacca ccccatttat ataggcggcc gcggatccgg cgcaacaaac
     4501 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgat ggccaagcct
     4561 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     4621 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     4681 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     4741 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     4801 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     4861 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     4921 ccctctggtt atgtgtggga gggctaagaa taattcgata tcaagcttat cggtaatcaa
     4981 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     5041 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     5101 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     5161 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     5221 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     5281 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     5341 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     5401 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     5461 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     5521 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     5581 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
     5641 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
     5701 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     5761 gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
     5821 taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc agggatcaga
     5881 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa
     5941 gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca tgggatggat
     6001 gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt tcatcacatg
     6061 gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg
     6121 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     6181 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     6241 ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac
     6301 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     6361 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     6421 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6481 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     6541 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     6601 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     6661 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     6721 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     6781 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     6841 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     6901 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6961 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     7021 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     7081 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     7141 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc cagttccgcc
     7201 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     7261 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     7321 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     7381 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     7441 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     7501 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     7561 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     7621 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     7681 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     7741 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     7801 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     7861 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7921 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     7981 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     8041 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     8101 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     8161 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     8221 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     8281 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     8341 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     8401 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     8461 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     8521 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     8581 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     8641 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     8701 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     8761 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     8821 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     8881 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     8941 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     9001 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     9061 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     9121 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     9181 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     9241 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     9301 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     9361 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     9421 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     9481 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     9541 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     9601 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     9661 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     9721 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     9781 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     9841 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     9901 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     9961 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
    10021 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
    10081 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
    10141 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
    10201 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
    10261 ac
//
