LOCUS       sequence-372945-       12389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TetON-APEX2-V5-BPNLS-dRfxCas13d-dsRBD-BPNLS-P2A-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12389)
  AUTHORS   Han S, Zhao BS, Myers SA, Carr SA, He C, Ting AY
  TITLE     RNA-protein interaction mapping via MS2- or Cas13-based APEX 
            targeting.
  JOURNAL   Proc Natl Acad Sci U S A. 2020 Aug 24. pii: 2006617117. doi: 
            10.1073/pnas.2006617117.
  PUBMED    32839320
REFERENCE   2  (bases 1 to 12389)
  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: "Proc Natl 
            Acad Sci U S A. 2020 Aug 24. pii: 2006617117. doi: 
            10.1073/pnas.2006617117."
COMMENT     Sequence Label: dCas13d-dsRBD-APEX2
FEATURES             Location/Qualifiers
     source          1..12389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     132..151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     385..402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    519..540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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)."
     primer_bind     598..620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1124..1158
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1921..2042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1932..1951)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1986..2005
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2075..2674)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2158..2178)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2655..2674
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(2675..2728)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(2729..3472)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     LTR             3506..3774
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(3787..3998)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    4229..4277
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4302..4321
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(4360..5073,5074..5076,5077..5079))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(4363..5073,5074..5076,5077..5079))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4405..4426)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4754..4773
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5013..5034
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(5086..5142)
                     /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             complement(5152..5172)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5461..8358)
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLAHWVVANNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFANKAVALEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             complement(8368..8388)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8428..8469)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(8470..9219)
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="MGKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAG
                     TFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVA
                     VEVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGH
                     TIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVD
                     KYAADEDAFFADYAEAHQKLSELGFADA"
     regulatory      9216..9225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(9238..9616)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     primer_bind     complement(9305..9329)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    9374..9392
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9410..9428
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9446..9464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9482..9500
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9518..9536
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9554..9572
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9590..9608
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     repeat_region   10065..10127
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10257..10274)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10257..10273)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10266..10288)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10415..10870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10502..10521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10712..10733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10896..11000
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11001..11069,11070..11861)
                     /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     complement(11219..11238)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(12032..12389,1..231)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
       61 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      121 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      181 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      241 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      301 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      361 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcccgccgg
      661 gtaactcacg gggtatccat gtccatttct gcggcatcca gccaggatac ccgtcctcgc
      721 tgacgtaata tcccagcgcc gcaccgctgt cattaatctg cacaccggca cggcagttcc
      781 ggctgtcgcc ggtattgttc gggttgctga tgcgcttcgg gctgaccatc cggaactgtg
      841 tccggaaaag ccgcgacgaa ctggtatccc aggtggcctg aacgaacagt tcaccgttaa
      901 aggcgtgcat ggccacacct tcccgaatca tcatggtaaa cgtgcgtttt cgctcaacgt
      961 caatgcagca gcagtcatcc tcggcaaact ctttccatgc cgcttcaacc tcgcgggaaa
     1021 aggcacgggc ttcttcctcc ccgatgccca gatagcgcca gcttgggcga tgactgagcc
     1081 ggaaaaaaga cccgacgata tgatcctgat gcagctagat taaccctaga aagatagtct
     1141 gcgtaaaatt gacgcatgca ttcttgaaat attgctctct ctttctaaat agcgcgaatc
     1201 cgtcgctgtg catttaggac atctcagtcg ccgcttggag ctcccgtgag gcgtgcttgt
     1261 caatgcggta agtgtcactg attttgaact ataacgaccg cgtgagtcaa aatgacgcat
     1321 gattatcttt tacgtgactt ttaagattta actcatacga taattatatt gttatttcat
     1381 gttctactta cgtgataact tattatatat atattttctt gttatagata tcagggacag
     1441 ccccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg
     1501 agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg
     1561 cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg
     1621 ctgctctttg agcctgcaga cacctggggg gatacgggga aaaagccata ccaatgggcc
     1681 ctaaaaaagg aatccagtca attccggggc taaacctggc tgccactgtt tctttaggga
     1741 cttcgttcct gtgaggacag gaaaccctgc aacctggttg accagagcaa gggatataag
     1801 aacccagttt gggaacactg ctgtttaaat attaaacagg gatgtcttgt cccccggaag
     1861 ggagaccaga tgagcttctg atagactaat tgacatcatt tgagtcaatt ccagacatga
     1921 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     1981 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2041 tttacccaca aattttgtaa tccagaggtt gatttcaggc accgggcttg cgggtcatgc
     2101 accaggtgcg cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct
     2161 cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct
     2221 cggccgcctc cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg
     2281 gcgagacgcc gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga
     2341 ggccttccat ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc
     2401 cgatctcggc gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg
     2461 cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga
     2521 gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg
     2581 cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg
     2641 tggcgaggcg caccgtgggc ttgtactcgg tcatagggcc gggattctcc tccacgtcac
     2701 cgcatgttag aagacttcct ctgccctccc cggggagcat gtcaaggtca aaatcgtcaa
     2761 gagcgtcagc aggcagcata tcaaggtcaa agtcgtcaag ggcatcggct gggagcatgt
     2821 ctaagtcaaa atcgtcaagg gcgtcggtcg gcccgccgct ttcgcacttt agctgtttct
     2881 ccaggccaca tatgattagt tccaggccga aaaggaaggc aggttcggct ccctgccggt
     2941 cgaacagctc aattgcttgt ttcagaagtg ggggcataga atcggtggta ggtgtctctc
     3001 tttcctcttt tgctacttga tgctcctgtt cctccaatac gcagcccagt gtaaagtggc
     3061 ccacggcgga cagagcgtac agtgcgttct ccagggagaa gccttgctga cacaggaacg
     3121 cgagctgatt ttccagggtt tcgtactgtt tctctgttgg gcgggtgccg agatgcactt
     3181 tagccccgtc gcgatgtgag aggagagcac agcggtatga cttggcgttg ttccgcagaa
     3241 agtcttgcca tgactcgcct tccagggggc aggagtgggt atgatgcctg tccagcatct
     3301 cgattggcag ggcatcgagc agggcccgct tgttcttcac gtgccagtac agggtaggct
     3361 gctcaactcc cagcttttga gcgagtttcc ttgtcgtcag gccttcgata ccgactccat
     3421 tgagtaattc cagagcagag tttatgactt tgctcttgtc cagtctagac atggcggctc
     3481 tagagtaggc gccggtcaca gcttagatct gtaacggcgc agaacagaaa acgaaacaaa
     3541 gacgtagagt tgagcaagca gggtcaggca aagcgtggag agccggctga gtctaggtag
     3601 gctccaaggg agcgccggac aaaggcccgg tctcgacctg agctttaaac ttacctagac
     3661 ggcggacgca gttcaggagg caccacaggc gggaggcggc agaacgcgac tcaaccggcg
     3721 tggatggcgg cctcaggtag ggcggcgggc gcgtgaagga gagatgcgag cccctcgaag
     3781 cttcagctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc
     3841 acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact
     3901 ttcccagttt accccgcgcc accttctcta ggcacccgtt caattgccga cccctccccc
     3961 caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac cggagcgatc
     4021 gcagatccac ttagattcag ctggatccga tttcgcagat ctgctatggc agggcctgcc
     4081 gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg gggtggggaa
     4141 aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc gacagagtgc
     4201 cagccctggg accgaacccc gcgtttatga acaaacgacc caacacccgt gcgttttatt
     4261 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     4321 agttagcctc ccccgtttaa actcattact aaccggttac tacttgtaca gctcgtccat
     4381 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     4441 gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac
     4501 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     4561 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
     4621 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
     4681 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     4741 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
     4801 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     4861 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     4921 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     4981 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     5041 caccaccccg gtgaacagct cctcgccctt gctcaccatg aattcaggtc cagggttctc
     5101 ctccacgtct ccagcctgct tcagcaggct gaagttagta gctccgcttc cgaccttgcg
     5161 ctttttcttg gggctttcga actcggaacc atcggcggtg cgcttggatc ccgctttctt
     5221 ttctttattg aggatttcga ctgccaattt agcggctgcg ttcttcgctt ctttcttact
     5281 tcggccttcg ccttctggga actctctacc atcgataatt acttgaaaag tgaatcttcg
     5341 gtcgtgaggt gggcctgagt tcggcaactc ttggtacttc agcacaactc cctgtttctg
     5401 gcgataagtg ttcagttctt ccatgaagaa acctgcggac aaatcgcctg ctccggatcc
     5461 ggaattgccg gacaccttct ttttctcctt gtcgaacttg gcggcctcgt tcctatcgaa
     5521 cagggcctcg atgctcaggt tcttaaacct ggggatacag tagccgaaag gcacacacag
     5581 cagtttcagg agcctatcgt tgtacttctt ctcgtcattc acagcgtcga agtactcgga
     5641 cacctttccg ctgcttttct cgtacctctc attcatgata attctctgca tgatgtaatg
     5701 gtacagttgg aagtaggaat tgacctcggc aatgtcgttg atataggcgt ggacatacct
     5761 ggccacttcc agggcgacgg ccttgtttgc gaacagggta catgtcttgt tgtcaattct
     5821 caggaggtcc tccctgatga tcacattcca cttggtgttc ctcaggtagg cgttcagggc
     5881 ggttttggcc ttctccctgt taatctgcct ggtgaactcc tcggctttct tctcgtcgct
     5941 gtatttgatg taattggcat acagcttggg gttggcgctc tcgaggctgt caatgctctc
     6001 cttggctctt tcagccatct ccttttccac gtcctttctc ttatcggggg cagtttcatc
     6061 aatgccagcg cagagcttgg tgacggagct gaatcccttc tcttccagtt tcttgagatt
     6121 gatgtcgtag cctttctcct tgtacagttg agcatcacgc tcgacgcaat ggaatccgat
     6181 gacgtacctg gcgttgatat tgacaatatt cttgaggatg tggtagatca cggtgaggta
     6241 caggctgatg atctttttaa acttctccct ctcggcgttt tccctgccgg tgtcctcaat
     6301 gacgctcctt ttcttgtcga attggtcgta gttcattccg gtgatgatct ttgtgagagc
     6361 gtccaccttt tcgctcacgc tcttgccctt atcctttccg atacaagttt cgtagtacct
     6421 gtcgatctgg ttcttgccgt tctggccctg ttttttctgg atgtcagcga tcctgccgag
     6481 cacgaacttc accacggcct cgtttttggc gatctcatgg aggtgggcag gatcaccgta
     6541 tctgatcagg tagtggaacc ttttattgct gatcacgtta ttaataatga aatttctcat
     6601 gccgtgcttg cctttcttga gcttgtttcc gttctcgtcc agggaaaagg tgtcggcgag
     6661 ggccttgagc tcatcatagg acaggttggt tcctaaaata cggatggcgt cgatatacat
     6721 ggccctcctg gcatcggcaa taggttctcc cattctagcg aaggacttga tcagcctcag
     6781 ctcatcggcg atcttggcgg agtctttgaa aaaggcgtat tcctccacga acttagcgtt
     6841 gactccgatg agaggcatca ccttcaggaa gctctggatg ttatcgaatt tattaatcag
     6901 ggtggtcagg aggtcgttga tctccttgcc atccaggaac atggtcaggg catacatgag
     6961 tttgctgaag gcggaaacat cacggccagc gggcaggatt ctgggcagtc taggggcgtc
     7021 cttcttctta tactctcttg tcttgtttcc cctaaattcc ttgatgttgt gcatgatatt
     7081 ttcgagcttt ctccaaattc tattagcttc atcgtagtag agggcatcct tctggtcgtc
     7141 gttgaaggag cccctcaggt taatcacaaa gatatccttc tcgctcaggg acttctcatt
     7201 atcggggagg gacttattgg cggcagccac cttggcatcc tcttcgatgt aatacctata
     7261 aatcacaaag tccatcatgg tgtagacctt ggtcctgatg gagtcgaaca ccttatgatt
     7321 tttcctgatc tcggacatat ccttcctgtc cagcatcact tccctgagct tggtgatatt
     7381 gaatccgagg tttttctgct ctttcataat gctgaatctg aaatattgtt cggcgaattc
     7441 ggcagggttg attcccagag tttcggcaat atagttcacg ttggcggcgg agttcttgga
     7501 gaaggagttg gtcagctcat tggtgatcct gtcgtagagg tagttgaggg tggagatgta
     7561 ttcgttgtcg aggttcttat cgaggttgta gagccaggtc ctggagatcc tggactcttc
     7621 ttcgttgtta gcgaccaccc agtgcgccag tccgctcagg agggccagaa tgtcatagca
     7681 ttcgttgccg taattgatga tgtaatttct gccctccttg ctgaaaaagg cctggccgaa
     7741 atagccgagt ctggggttat cgaggaagtt gtcgaactcg tcatactggg ccttgatggc
     7801 gttgatgagc ttatcgttat tgttgaaagc ggccctatgg tgctcggggt ctttgaattc
     7861 gtcgtaggta tacactgtgg agaacttgcc gaatccaata atgtccttat ccaggccgga
     7921 gatattgttg acggcgtagg cggcgttggt aatgtattcg gcgaggattt tttcaatgtc
     7981 caggatgtta tggatcacct ggatacaaat attgtcattg ccatcagcgc tctcgccgaa
     8041 gtacctcttt tccagagttt ccttcaggcc gagcatatcc tgctggacgg gtcctgtata
     8101 cagagggttg ttagccacca cggcgtagcc cttaggatgg ctgaatttgg cgttgccgat
     8161 cttatagccg gcgtttttat cggccatttc agcgctgaag gcctcgccct cattcacgct
     8221 cctgatgctg tcgccctcca cgatcttttc cagcctggcg tcgctgcctt cggcgaaggt
     8281 tgtcatgtac actttggagc cggacacgag tgtggacttc acgcccatgc ccttggcgaa
     8341 ggactttttt ttttcgatgc tggcctccac ctttctcttc ttcttggggc tctcaaattc
     8401 actgccatca gcagtccgct tgctaccggt gctgtccagg cccagcaggg ggttggggat
     8461 gggcttgccg gcatcagcaa acccaagctc ggaaagcttt tggtgagcct cagcgtaatc
     8521 agcaaagaag gcatcttcgt ccgctgcata tttgtcaacg agagggcgga atacagggtc
     8581 agacaaaaga gccttgtcag aaggtagctg aaggagacct tccttctcac cactcaacaa
     8641 ctccgtgaag tatgagttgt cgaaaataag aggattagag gtccagggac cctcaaatcc
     8701 agaacgctcc ttgtgtgcag ctccaatagt gtgaccccca gatagagcaa cgatatcttg
     8761 gtcagtaagc cccatagctt tgccaaacac atctctcaaa tggtcagaac ccttagtggg
     8821 atcgggcaag cgaccctctg gtggtggctc aggcttgtcc tctcttccag ggtggaatgg
     8881 aaccttaggt ccacccgtga cctcaacggc aacaacgcca gccaactggt agaaatcggc
     8941 gtagctcaaa atagggaact ccgccttgag tggctccaaa agcctaacag cgatgtcaag
     9001 accgttgtta gcgctgtgag ccagttcggc agggtgcttg atggttccga agggtccacc
     9061 ggtcttcgtg cccttgtcaa aggttccagc agagtggaat gccaaacgga gcattagagg
     9121 agcgcatctc ttctcagcga tgaagcctct gagcttcttc ttcgccttct caacggcgtc
     9181 ctggtaatca gcactcacag ttgggtaaga ctttcccatg gtggctagct ctagaccttt
     9241 acgagggtag gaagtggtac ggaaagttgg tataagacaa aagtgttgtg gaattgctcc
     9301 aggcgatctg acggttcact aaacgagctc tgcttttata ggcgcccacc gtacacgcct
     9361 aaagcttata cgttctctat cactgatagg gagtaaactg gatatacgtt ctctatcact
     9421 gatagggagt aaactgtaga tacgttctct atcactgata gggagtaaac tggtcatacg
     9481 ttctctatca ctgataggga gtaaactcct tatacgttct ctatcactga tagggagtaa
     9541 agtctgcata cgttctctat cactgatagg gagtaaactc ttcatacgtt ctctatcact
     9601 gatagggagt aaactcgagg aattcttgaa gacgaaagac tagtggccgt ggaggccttt
     9661 tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa
     9721 gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat
     9781 gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg
     9841 ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctca taaaagtttt
     9901 gttactttat agaagaaatt ttgagttttt gtttttttta ataaataaat aaacataaat
     9961 aaattgtttg ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct
    10021 attcaaatta ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac
    10081 acatgattat ctttaacgta cgtcacaata tgattatctt tctagggtta atctagctgc
    10141 gtgttctgca gcgtgtcgag catcttcatc tgctccatca cgctgtaaaa cacatttgca
    10201 ccgcgagtct gcccgtcctc cacgggttca aaaacgtgaa tgaacgaggc gcgctcactg
    10261 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
    10321 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
    10381 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
    10441 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
    10501 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
    10561 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
    10621 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
    10681 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
    10741 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
    10801 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
    10861 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
    10921 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
    10981 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
    11041 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
    11101 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
    11161 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
    11221 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
    11281 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
    11341 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
    11401 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
    11461 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
    11521 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
    11581 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
    11641 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
    11701 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
    11761 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
    11821 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
    11881 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
    11941 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
    12001 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
    12061 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
    12121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
    12181 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
    12241 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
    12301 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
    12361 cgtgcacaca gcccagcttg gagcgaacg
//
