LOCUS       sequence-333012-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Disrupt IREB2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Terzi EM, Sviderskiy VO, Alvarez SW, Whiten GC, Possemato R
  TITLE     Iron-sulfur cluster deficiency can be sensed by IRP2 and regulates 
            iron homeostasis and sensitivity to ferroptosis independent of IRP1 
            and FBXL5.
  JOURNAL   Sci Adv. 2021 May 26;7(22). pii: 7/22/eabg4302. doi: 
            10.1126/sciadv.abg4302. Print 2021 May.
  PUBMED    34039609
REFERENCE   2  (bases 1 to 13013)
  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 Adv."; 
            date: "2021-05-26"; volume: "7(22). pii: 7/22/eabg4302. doi"; pages:
            " 10.1126/sciadv.abg4302. Print 2021 May"
COMMENT     Sequence Label: pLentiCRISPR V2 sgIREB2_1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..45)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(199..787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(279..298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(958..1749,1750..1818))
                     /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     1581..1600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1819..1923)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1998..2017)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2189..2568
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2570..2768
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2723..2743
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2786..2966
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3013..3138
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3631..3864
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4049..4093
                     /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    4391..4508
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4559..4807
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4559..4579
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4730..4749
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4829..4904
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4966..5177
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5196..5205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5202..9305
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5202..9305
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9306..9353
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9354..9377
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9387..9443
                     /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             9444..10040
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9937..9957
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10056..10644
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10109..10129)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10527..10538)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10716..10949
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10975..10992)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10981..11205
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11251..11679
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11338..11357)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11548..11569
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11688..11708)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11693..12022
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11873..12008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11935..11954
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12070..12117
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12136..12510
                     /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    12640..12761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12677..12696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12731..12750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12810..12826)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12810..12826)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12823..12845)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12834..12850
                     /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(12858..12864,12865..12882,12883..12888))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12903..12924
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
       61 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      601 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1921 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     1981 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     2041 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     2101 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     2161 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     2221 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     2281 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     2341 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     2401 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     2461 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     2521 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     2581 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     2641 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2701 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     2761 ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
     2821 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     2881 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     2941 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     3001 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     3061 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     3121 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     3181 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     3241 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     3301 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     3361 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     3421 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     3481 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     3541 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     3601 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     3661 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     3721 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     3781 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     3841 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     3901 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     3961 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     4021 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     4081 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     4141 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     4201 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     4261 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     4321 cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat
     4381 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     4441 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     4501 aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt aaggtaccga
     4561 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     4621 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     4681 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     4741 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     4801 aaacaccgaa tgcaccaaat cctggagggt tttagagcta gaaatagcaa gttaaaataa
     4861 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gaattcgcta
     4921 gctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     4981 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
     5041 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     5101 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     5161 ttgccgccag aacacaggac cggttctaga gcgctgccac catggacaag aagtacagca
     5221 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     5281 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     5341 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     5401 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     5461 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     5521 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     5581 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     5641 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     5701 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     5761 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     5821 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     5881 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
     5941 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     6001 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     6061 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     6121 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     6181 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     6241 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     6301 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     6361 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     6421 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     6481 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     6541 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     6601 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     6661 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     6721 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     6781 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     6841 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     6901 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     6961 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     7021 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     7081 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     7141 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     7201 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     7261 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     7321 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     7381 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     7441 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     7501 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     7561 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     7621 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     7681 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     7741 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     7801 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     7861 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     7921 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     7981 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     8041 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     8101 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     8161 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     8221 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     8281 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     8341 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     8401 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     8461 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     8521 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     8581 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     8641 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     8701 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     8761 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     8821 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     8881 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     8941 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     9001 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     9061 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     9121 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     9181 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     9241 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     9301 gcgacaagcg acctgccgcc acaaagaagg ctggacaggc taagaagaag aaagattaca
     9361 aagacgatga cgataaggga tccggcgcaa caaacttctc tctgctgaaa caagccggag
     9421 atgtcgaaga gaatcctgga ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     9481 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     9541 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     9601 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     9661 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     9721 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     9781 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc
     9841 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     9901 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     9961 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
    10021 cccgcaagcc cggtgcctga acgcgttaag tcgacaatca acctctggat tacaaaattt
    10081 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
    10141 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
    10201 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
    10261 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
    10321 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
    10381 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
    10441 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
    10501 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
    10561 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
    10621 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
    10681 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
    10741 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
    10801 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    10861 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    10921 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
    10981 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
    11041 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
    11101 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
    11161 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
    11221 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
    11281 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    11341 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    11401 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    11461 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    11521 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    11581 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    11641 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
    11701 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    11761 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    11821 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
    11881 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    11941 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
    12001 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
    12061 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    12121 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    12181 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    12241 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    12301 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    12361 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    12421 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    12481 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    12541 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    12601 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    12661 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    12721 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    12781 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    12841 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    12901 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
    12961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cgg
//
