LOCUS       sequence-291733-       15341 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for an inducible BirA-NRF2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15341)
  AUTHORS   Pereira EJ, Burns JS, Lee CY, Marohl T, Calderon D, Wang L, Atkins 
            KA, Wang CC, Janes KA
  TITLE     Sporadic activation of an oxidative stress-dependent NRF2-p53 
            signaling network in breast epithelial spheroids and 
            premalignancies.
  JOURNAL   Sci Signal. 2020 Apr 14;13(627). pii: 13/627/eaba4200. doi: 
            10.1126/scisignal.aba4200.
  PUBMED    32291314
REFERENCE   2  (bases 1 to 15341)
  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: "Sci 
            Signal."; date: "2020-04-14"; volume: "13(627). pii: 
            13/627/eaba4200. doi"; pages: " 10.1126/scisignal.aba4200"
COMMENT     Sequence Label: pSLIK_hygro_BirA_NRF2
FEATURES             Location/Qualifiers
     source          1..15341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2377..2401
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    2441..2459
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2477..2495
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2513..2531
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2549..2567
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2579..2617
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2614..2638
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2648..2998,2999..3001,3002..3610)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     protein_bind    complement(5467..5491)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5541..6752
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6589..6608
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(6877..7494,7495..7500,7501..7536,7537..7539,7540..7575,
                     7576..7578,7579..7614,7615..7623)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    7786..8248
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7842..7859)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8069..8088
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8307..9326
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             8307..9326
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    9792..10380
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9845..9865)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10263..10274)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10383..10399)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10384..10400)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10909..11089
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11115..11132)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11121..11345
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11391..11819
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11478..11497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11688..11709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11828..11848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11833..12162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12013..12148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12075..12094
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12210..12257
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12276..12650
                     /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    12780..12901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12817..12836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12871..12890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12950..12966)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12950..12966)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12963..12985)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12974..12990
                     /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(12998..13004,13005..13022,13023..13028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13043..13064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13181..13198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13352..13940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13432..13451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14111..14902,14903..14971))
                     /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     14734..14753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14972..15076)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(15151..15170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttctagacag ttatcaacaa gtttgtacaa aaaagcaggc
     2401 tttaaaggaa ccaattcagt cgactggatc ccgagtttac tccctatcag tgatagagaa
     2461 cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc
     2521 agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagg
     2581 taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc
     2641 cactagaatg aaggacaaca ccgtgcccct gaagctgatc gccctgctgg ccaacggcga
     2701 gttccactct ggcgagcagc tgggagagac cctgggaatg agcagagccg ccatcaacaa
     2761 gcacatccag acactgagag actggggagt ggacgtgttc accgtgcctg gcaagggcta
     2821 cagcctgcct gagcctatcc agctgctgaa cgccaagcag atcctgggac agctggatgg
     2881 cggaagcgtg gccgtgctgc ctgtgatcga ctccaccaat cagtacctgc tggacagaat
     2941 cggagagctg aagtccggcg acgcctgcat cgccgagtac cagcaggctg gcagaggagg
     3001 cagaggacgg aagtggttca gcccattcgg agccaacctg tacctgtcca tgttctggag
     3061 actggagcag ggacctgctg ctgccatcgg actgagtctg gtgatcggaa tcgtgatggc
     3121 cgaggtgctg agaaagctgg gagccgacaa ggtgagagtg aagtggccta atgacctgta
     3181 cctccaggac cgcaagctgg ctggcatcct ggtggagctg acaggcaaga caggcgatgc
     3241 cgctcagatc gtgatcggag ccggaatcaa catggccatg agaagagtgg aggagagcgt
     3301 ggtgaaccag ggctggatca ccctgcagga ggctggcatc aacctggacc ggaacaccct
     3361 ggccgccatg ctgatcagag agctgagagc cgctctggag ctgttcgagc aggagggact
     3421 ggctccttac ctgagcagat gggagaagct ggacaacttc atcaacagac ctgtgaagct
     3481 gatcatcggc gacaaggaaa tcttcggcat ctccagagga atcgacaagc agggagctct
     3541 gctgctggag caggacggaa tcatcaagcc ctggatgggc ggagaaatct ccctgagaag
     3601 cgcagagaag actagtatgg acttggagct gccgccgccg ggactcccgt cccagcagga
     3661 catggatttg attgacatac tttggaggca agatatagat cttggagtaa gtcgagaagt
     3721 atttgacttc agtcagcgac ggaaagagta tgagctggaa aaacagaaaa aacttgaaaa
     3781 ggaaagacaa gaacaactcc aaaaggagca agagaaagcc tttttcgctc agttacaact
     3841 agatgaagag acaggtgaat ttctcccaat tcagccagcc cagcacatcc agtcagaaac
     3901 cagtggatct gccaactact cccaggttgc ccacattccc aaatcagatg ctttgtactt
     3961 tgatgactgc atgcagcttt tggcgcagac attcccgttt gtagatgaca atgaggtttc
     4021 ttcggctacg tttcagtcac ttgttcctga tattcccggt cacatcgaga gcccagtctt
     4081 cattgctact aatcaggctc agtcacctga aacttctgtt gctcaggtag cccctgttga
     4141 tttagacggt atgcaacagg acattgagca agtttgggag gagctattat ccattcctga
     4201 gttacagtgt cttaatattg aaaatgacaa gctggttgag actaccatgg ttccaagtcc
     4261 agaagccaaa ctgacagaag ttgacaatta tcatttttac tcatctatac cctcaatgga
     4321 aaaagaagta ggtaactgta gtccacattt tcttaatgct tttgaggatt ccttcagcag
     4381 catcctctcc acagaagacc ccaaccagtt gacagtgaac tcattaaatt cagatgccac
     4441 agtcaacaca gattttggtg atgaatttta ttctgctttc atagctgagc ccagtatcag
     4501 caacagcatg ccctcacctg ctactttaag ccattcactc tctgaacttc taaatgggcc
     4561 cattgatgtt tctgatctat cactttgcaa agctttcaac caaaaccacc ctgaaagcac
     4621 agcagaattc aatgattctg actccggcat ttcactaaac acaagtccca gtgtggcatc
     4681 accagaacac tcagtggaat cttccagcta tggagacaca ctacttggcc tcagtgattc
     4741 tgaagtggaa gagctagata gtgcccctgg aagtgtcaaa cagaatggtc ctaaaacacc
     4801 agtacattct tctggggata tggtacaacc cttgtcacca tctcaggggc agagcactca
     4861 cgtgcatgat gcccaatgtg agaacacacc agagaaagaa ttgcctgtaa gtcctggtca
     4921 tcggaaaacc ccattcacaa aagacaaaca ttcaagccgc ttggaggctc atctcacaag
     4981 agatgaactt agggcaaaag ctctccatat cccattccct gtagaaaaaa tcattaacct
     5041 ccctgttgtt gacttcaacg aaatgatgtc caaagagcag ttcaatgaag ctcaacttgc
     5101 attaattcgg gatatacgta ggaggggtaa gaataaagtg gctgctcaga attgcagaaa
     5161 aagaaaactg gaaaatatag tagaactaga gcaagattta gatcatttga aagatgaaaa
     5221 agaaaaattg ctcaaagaaa aaggagaaaa tgacaaaagc cttcacctac tgaaaaaaca
     5281 actcagcacc ttatatctcg aagttttcag catgctacgt gatgaagatg gaaaacctta
     5341 ttctcctagt gaatactccc tgcagcaaac aagagatggc aatgttttcc ttgttcccaa
     5401 aagtaagaag ccagatgtta agaaaaacta gcaattggaa gactaatgcg ttttcgagat
     5461 atctagaccc agctttcttg tacaaagtgg ttgataacat ctcgagccat aaagatggtt
     5521 aattaaccca cccaagatct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     5581 cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc
     5641 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     5701 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     5761 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     5821 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     5881 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     5941 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     6001 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     6061 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     6121 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     6181 acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     6241 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     6301 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     6361 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     6421 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     6481 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     6541 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     6601 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     6661 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta
     6721 aattctggcc gtttttggct tttttgttag acgaagcttg gtaccgagct cggatctcca
     6781 ccccgtaccg gtcctgcagg atttaaatgg cgcgccgcta gcgctaccgg actcagatct
     6841 cgagctcaag cttcgaattc tgcagtcgaa ttcaccatgt ctagactgga caagagcaaa
     6901 gtcataaacg gagctctgga attactcaat ggtgtcggta tcgaaggcct gacgacaagg
     6961 aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag
     7021 cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc
     7081 cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc ataccgctgt
     7141 gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa
     7201 cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag
     7261 aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa
     7321 caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca
     7381 cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc
     7441 ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg
     7501 accgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt
     7561 gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg
     7621 taactaagta aggatccgcg gccgcactag aggaattccg cccctctccc cccccccntc
     7681 tcccnnnnnn ncccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     7741 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     7801 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     7861 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     7921 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     7981 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     8041 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     8101 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     8161 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     8221 ggttttcctt tgaaaaacac gatgataagc ttaccggtac gcgtgatagc ttgccacaac
     8281 ccgtaccaaa gatggataga tccggaaagc ctgaactcac cgcgacgtct gtcgagaagt
     8341 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     8401 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     8461 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     8521 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     8581 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     8641 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     8701 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     8761 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     8821 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     8881 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     8941 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     9001 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     9061 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     9121 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     9181 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     9241 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     9301 gcactcgtcc gagggcaaag gaatagagta gatgccgacc gaacaagagc tgatttcgag
     9361 aacgcctcag ccagcaactc gcgcgagcct agcaaggcaa atgcgagaga acggccttac
     9421 gcttggtggc acagttctcg tccacagttc gctaagctcg ctcggctggg tcgcgggagg
     9481 gccggtcgca gtgattcagg cccttctgga ttgtgttggt ccccagggca cgattgtcat
     9541 gcccacgcac tcgggtgatc tgactgatcc cgcagattgg agatcgccgc ccgtgcctgc
     9601 cgattgggtg cagatctcta gatctgtaca agtaaagcgg ccgcgactct agatcataat
     9661 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     9721 gaacctgaaa cataaaatga atgcaattgt tgttgtttag tccctcccaa ttcgatatca
     9781 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     9841 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     9901 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     9961 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
    10021 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
    10081 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
    10141 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
    10201 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
    10261 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
    10321 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
    10381 atcgataccg tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat
    10441 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt
    10501 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
    10561 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
    10621 gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac
    10681 acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca
    10741 gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct
    10801 gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc
    10861 cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg
    10921 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
    10981 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
    11041 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
    11101 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
    11161 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
    11221 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
    11281 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
    11341 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
    11401 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
    11461 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
    11521 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
    11581 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
    11641 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
    11701 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
    11761 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
    11821 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
    11881 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
    11941 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
    12001 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
    12061 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
    12121 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
    12181 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
    12241 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
    12301 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
    12361 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
    12421 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
    12481 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
    12541 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
    12601 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
    12661 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
    12721 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
    12781 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    12841 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    12901 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
    12961 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    13021 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    13081 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    13141 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    13201 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    13261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    13321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    13381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    13441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    13501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    13561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    13621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    13681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    13741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    13801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    13861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    13921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    13981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    14041 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    14101 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    14161 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    14221 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    14281 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    14341 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    14401 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    14461 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    14521 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    14581 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    14641 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    14701 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    14761 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    14821 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    14881 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    14941 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    15001 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    15061 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    15121 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
    15181 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
    15241 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
    15301 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
