LOCUS       Exported               16277 bp ds-DNA     circular SYN 19-DEC-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 16277)
  AUTHORS   sidhartm
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Dec 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16277
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     CDS             327..986
                     /codon_start=1
                     /label=AmpR
                     /note="AmpR"
                     /note="color: #99ff33"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      1137..1804
                     /label=pUC Ori
                     /note="pUC Ori"
                     /note="color: #ffff99"
     primer_bind     1649..1668
                     /label=pBR322ori-FSEQ
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     misc_recomb     2208..3044
                     /label=HA-R
                     /note="HA-R"
                     /note="color: #dd0000"
     primer_bind     2890..2910
                     /label=nipun fwd
                     /note="color: black; sequence: gttctggcaaggagagagatg"
     promoter        3074..3389
                     /note="TRE"
                     /note="color: #0000dd; direction: RIGHT"
     promoter        3075..3389
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    3083..3101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    3119..3137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    3154..3172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    3190..3208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    3226..3244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    3261..3279
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    3297..3315
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     primer_bind     3362..3386
                     /label=435R
                     /note="color: black; sequence: AGCTCGTTTAGTGAACCGTCAGATC; 
                     added: 2022-03-22"
     primer_bind     3369..3390
                     /label=seq primerprom
                     /note="color: black; sequence: TTAGTGAACCGTCAGATCGCCT"
     misc_recomb     3406..3430
                     /label=AttB1
                     /note="AttB1"
                     /note="color: #000000"
     primer_bind     3414..3434
                     /label=292F
                     /note="color: black; sequence: GTACAAAAAAGCAGGCTCCGC; 
                     added: 2021-08-25"
     primer_bind     complement(3435..3465)
                     /label=421R
                     /note="color: black; sequence: 
                     AAGGAAAAAAGCGGCCGCGCACCGGTCCAACCTGAAAACC; added: 
                     2021-11-22"
     primer_bind     3465..3484
                     /label=T7fwSEQ
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     primer_bind     3465..3484
                     /label=T7
                     /note="T7 promoter, forward primer"
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     promoter        3465..3483
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3533..3553
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             3554..7654
                     /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"
                     /note="color: #993366"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     7686..7717
                     /label=455F (Fragment 1.FOR)
                     /note="color: black; sequence: 
                     ccgccgacggctctgaattcgagagccctaag; added: 2022-01-25"
     CDS             7712..7732
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             9833..9853
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             9863..9889
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /note="color: #993366"
                     /translation="PAAKRVKLD"
     CDS             9899..9955
                     /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."
                     /note="color: #cc99b2
                     Cleavage site after base 9952"
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(9935..9963)
                     /label=456R (Fragment 1.REV)
                     /note="color: black; sequence: 
                     GGCTCcatcggtccaggattctcttcgac; added: 2022-01-25"
     primer_bind     9949..9973
                     /label=457F (Fragment 2.FOR)
                     /note="color: black; sequence: tggaccgatgGAGCCTCACCACGAG; 
                     added: 2022-01-25"
     misc_feature    9956..9958
                     /label=Kozak sequence
                     /note="color: #a6acb3"
     CDS             9959..10261
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53DD
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /note="color: #993366"
                     /translation="EPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERF
                     EMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     misc_feature    9959..10045
                     /label=NLS
                     /note="color: #a6acb3"
     misc_feature    9977..10261
                     /label=hP53DD part based on addgene 25989
                     /note="color: #ffff00"
     misc_feature    10046..10159
                     /label=Tet.
                     /note="color: #0000ff"
     primer_bind     10046..10083
                     /label=442F (P53DD-Tet-F)
                     /note="P53DD-Tet-F"
                     /note="color: black; sequence: 
                     tccCCCGGGgccaccatgCTGGATGGAGAATATTTCACCCTTCAGATCCGTGGGCG; 
                     added: 2021-12-20"
     primer_bind     complement(10139..10159)
                     /label=444R (Tet-rev)
                     /note="color: black; sequence: 
                     AAGGAAAAAAGCGGCCGCGGCGCGCCtcaCCCTGGCTCCTTCCCAGCCTG; added: 
                     2021-12-20"
     misc_feature    10160..10261
                     /label=3' terminal
                     /note="color: #a6acb3"
     primer_bind     10254..10273
                     /label=501F
                     /note="color: black; sequence: CAGACTGAccggtgtttaGG; added:
                     2022-03-22"
     primer_bind     complement(10263..10279)
                     /label=458R (Fragment 2.REV)
                     /note="color: black; sequence: 
                     tggtgatggtgatgatgaccggtGGCGCGCCtaaacaccg; added: 
                     2022-01-25"
     primer_bind     complement(10263..10279)
                     /label=Fragment 2.REV
                     /note="color: black; sequence: 
                     tggtgatggtgatgatgaccggtGGCGCGCCtaaacaccg; added: 
                     2022-01-25"
     misc_feature    10280..10866
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="This feature has 3 segments:
                        1: 10280 .. 10854 / #a6acb3
                        2: 10855 .. 10857 / #a6acb3 / ATG
                        3: 10858 .. 10866 / #a6acb3"
     misc_feature    10870..13128
                     /label=MLH1dn
                     /note="color: #a6acb3"
     primer_bind     11015..11034
                     /label=503F
                     /note="color: black; sequence: tcgtgaaggagggcggactg; added:
                     2022-03-22"
     primer_bind     complement(11201..11220)
                     /label=547R
                     /note="color: black; sequence: tttggtggtgatggtcacat; added:
                     2022-04-05"
     primer_bind     11800..11819
                     /label=504F
                     /note="color: black; sequence: cacgaggtgcacttcctgca; added:
                     2022-03-22"
     primer_bind     12790..12809
                     /label=506F
                     /note="color: black; sequence: ctgatcgataactacgtgcc; added:
                     2022-03-22"
     primer_bind     complement(13109..13148)
                     /label=502R
                     /note="color: black; sequence: 
                     AAGAAAGCTGGGTCGGCGCGttagaaaaccttataaaggt; added: 
                     2022-03-22"
     primer_bind     13129..13142
                     /label=420F
                     /note="color: black; sequence: GGTGGGCGCGCCGACCCAGC; added:
                     2021-11-22"
     misc_recomb     13136..13160
                     /label=AttB2
                     /note="AttB2"
                     /note="color: #000000"
     primer_bind     13151..13170
                     /label=PolyA-F
                     /note="color: black; sequence: ACAAAGTGGTTGATCCGGGG"
     primer_bind     complement(13181..13200)
                     /label=505R
                     /note="color: black; sequence: AATTCTGCAGATAAACGCTG; added:
                     2022-03-22"
     polyA_signal    13216..13409
                     /label=pA
                     /note="pA"
                     /note="color: #0099ff; direction: RIGHT"
     polyA_signal    complement(13410..13637)
                     /label=pA
                     /note="pA"
                     /note="color: #ffffcc; direction: LEFT"
     CDS             complement(13672..14697)
                     /codon_start=1
                     /label=Hygro
                     /note="Hygro"
                     /note="color: #999900"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    14707..14769
                     /label=2A
                     /note="2A"
                     /note="color: #ffff00"
     misc_feature    14785..14809
                     /label=SA
                     /note="SA"
                     /note="color: #33cc00"
     misc_recomb     14816..15619
                     /label=HA-L
                     /note="HA-L"
                     /note="color: #dd0000"
     primer_bind     14847..14866
                     /label=HA-L-F
                     /note="color: black; sequence: AGTACCCAGAACCAGAGCCA"
     primer_bind     complement(14872..14892)
                     /label=near reverse left
                     /note="color: black; sequence: ccttatattcccagggccggt"
     primer_bind     complement(14952..14978)
                     /label=TEST4
                     /note="color: black; sequence: TTCTTCCTCCAACCCGGGCCCCTATGT"
     primer_bind     complement(15633..15652)
                     /label=T7fwSEQ
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     primer_bind     complement(15633..15652)
                     /label=T7
                     /note="T7 promoter, forward primer"
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     rep_origin      15837..16143
                     /direction=RIGHT
                     /label=f1(+) Ori
                     /note="f1(+) Ori"
                     /note="color: #ffff99"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctcaga gcagagccag
     2221 gaacccctgt agggaagggg caggagagcc aggggcatga gatggtggac gaggaagggg
     2281 gacagggaag cctgagcgcc tctcctgggc ttgccaagga ctcaaaccca gaagcccaga
     2341 gcagggcctt agggaagcgg gaccctgctc tgggcggagg aatatgtccc agatagcact
     2401 ggggactctt taaggaaaga aggatggaga aagagaaagg gagtagaggc ggccacgacc
     2461 tggtgaacac ctaggacgca ccattctcac aaagggagtt ttccacacgg acacccccct
     2521 cctcaccaca gccctgccag gacggggctg gctactggcc ttatctcaca ggtaaaactg
     2581 acgcacggag gaacaatata aattggggac tagaaaggtg aagagccaaa gttagaactc
     2641 aggaccaact tattctgatt ttgtttttcc aaactgcttc tcctcttggg aagtgtaagg
     2701 aagctgcagc accaggatca gtgaaacgca ccagacagcc gcgtcagagc agctcaggtt
     2761 ctgggagagg gtagcgcagg gtggccactg agaaccgggc aggtcacgca tccccccctt
     2821 ccctcccacc ccctgccaag ctctccctcc caggatcctc tctggctcca tcgtaagcaa
     2881 accttagagg ttctggcaag gagagagatg gctccaggaa atgggggtgt gtcaccagat
     2941 aaggaatctg cctaacagga ggtgggggtt agacccaata tcaggagact aggaaggagg
     3001 aggcctaagg atggggcttt tctgtcacca atcctgtccc tagtttaatt aaccctctag
     3061 agcttagtac tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc
     3121 cctatcagtg atagagaacg atgtcgagtt tactccctat cagtgataga gaacgtatgt
     3181 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
     3241 gagaacgtat gtcgagttta tccctatcag tgatagagaa cgtatgtcga gtttactccc
     3301 tatcagtgat agagaacgta tgtcgaggta ggcgtgtacg gtgggaggcc tatataagca
     3361 gagctcgttt agtgaaccgt cagatcgcct ggagaattcc gatcaacaag tttgtacaaa
     3421 aaagcaggct ccgcggtttt caggttggac cggtgcgcgg ccgctaatac gactcactat
     3481 agggagagcc gccaccatga aacggacagc cgacggaagc gagttcgagt caccaaagaa
     3541 gaagcggaaa gtcgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     3601 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     3661 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     3721 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     3781 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     3841 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     3901 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     3961 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     4021 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     4081 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     4141 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     4201 caagagcaga aagctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     4261 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     4321 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     4381 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     4441 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     4501 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     4561 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     4621 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     4681 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga agagagagga
     4741 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     4801 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     4861 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     4921 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     4981 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     5041 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     5101 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     5161 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     5221 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     5281 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     5341 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     5401 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     5461 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     5521 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     5581 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     5641 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     5701 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     5761 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     5821 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     5881 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     5941 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     6001 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     6061 tgtggacgct atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     6121 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     6181 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     6241 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     6301 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     6361 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     6421 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     6481 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     6541 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     6601 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     6661 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     6721 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     6781 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     6841 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     6901 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     6961 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     7021 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     7081 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     7141 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     7201 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     7261 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     7321 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     7381 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     7441 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     7501 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     7561 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     7621 gacacggatc gacctgtctc agctgggagg tgactccggc ggaagctctg gtggcagcaa
     7681 gcggaccgcc gacggctctg aattcgagag ccctaagaag aaaagaaagg tgagcggagg
     7741 ctctagcggc ggaagcaccc tgaacattga agacgagtat agactgcatg aaacaagcaa
     7801 ggaacccgac gtgtccctgg gctccacctg gctgtccgac tttccccagg cctgggccga
     7861 gacaggagga atgggcctgg ccgtgcggca ggcacccctg atcatccctc tgaaggccac
     7921 ctctacaccc gtgagcatca agcagtaccc tatgtctcag gaggccagac tgggcatcaa
     7981 gcctcacatc cagaggctgc tggaccaggg catcctggtg ccatgccaga gcccctggaa
     8041 cacaccactg ctgcccgtga agaagccagg caccaatgac tatagacccg tgcaggatct
     8101 gagagaggtg aacaagaggg tggaggatat ccaccccacc gtgcccaacc cttacaatct
     8161 gctgtccggc ctgccccctt ctcaccagtg gtatacagtg ctggacctga aggatgcctt
     8221 cttttgtctg agactgcacc ctaccagcca gccactgttc gcctttgagt ggagggaccc
     8281 tgagatgggc atctctggcc agctgacctg gacacgcctg cctcagggct tcaagaatag
     8341 cccaacactg tttaacgagg ccctgcaccg cgacctggca gatttccgga tccagcaccc
     8401 agatctgatc ctgctgcagt acgtggacga tctgctgctg gccgccacca gcgagctgga
     8461 ttgccagcag ggaacacgcg ccctgctgca gaccctggga aacctgggat atagggcatc
     8521 cgccaagaag gcccagatct gtcagaagca ggtgaagtac ctgggctatc tgctgaagga
     8581 gggccagaga tggctgacag aggccaggaa ggagacagtg atgggccagc caacacccaa
     8641 gaccccaaga cagctgaggg agttcctggg caaagcagga ttttgcaggc tgttcatccc
     8701 aggattcgca gagatggcag cacctctgta cccactgacc aagccgggca ccctgtttaa
     8761 ttggggccct gaccagcaga aggcctatca ggagatcaag caggccctgc tgacagcacc
     8821 agccctgggc ctgccagacc tgaccaagcc tttcgagctg tttgtggatg agaagcaggg
     8881 ctacgccaag ggcgtgctga cccagaagct gggaccatgg agacggcccg tggcctatct
     8941 gtccaagaag ctggacccag tggcagcagg atggccacca tgcctgagga tggtggcagc
     9001 aatcgccgtg ctgacaaagg atgccggcaa gctgaccatg ggacagccac tggtcatcct
     9061 ggcaccacac gcagtggagg ccctggtgaa gcagcctcca gatcgctggc tgtctaacgc
     9121 ccggatgaca cactaccagg ccctgctgct ggacaccgat cgcgtgcagt ttggccctgt
     9181 ggtggccctg aatccagcca ccctgctgcc tctgccagag gagggcctgc agcacaactg
     9241 tctggacatc ctggcagagg cacacggaac aaggccagac ctgaccgatc agcccctgcc
     9301 tgacgccgat cacacatggt ataccgatgg aagctccctg ctgcaggagg gccagaggaa
     9361 ggcaggagca gcagtgacca cagagacaga agtgatctgg gccaaggccc tgccagcagg
     9421 cacatccgcc cagcgggccg agctgatcgc cctgacccag gccctgaaga tggccgaggg
     9481 caagaagctg aacgtgtaca cagactccag atatgccttc gccaccgcac acatccacgg
     9541 agagatctac aggcgccggg gctggctgac ctctgagggc aaggagatca agaacaagga
     9601 tgagatcctg gccctgctga aggccctgtt tctgcccaag cggctgagca tcatccactg
     9661 tcctggacac cagaagggac actccgccga ggcaaggggc aatcggatgg ccgaccaggc
     9721 cgccagaaag gctgctatta ctgaaactcc cgacacttcc actctgctga ttgaaaactc
     9781 ctccccttct ggcggctcaa aaagaaccgc cgacggcagc gaattcgagt ctcccaagaa
     9841 gaagaggaaa gtcggctctg gccctgccgc taagagagtg aagctggacg gatccggcgc
     9901 aacaaacttc tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgatgga
     9961 gcctcaccac gagctgcccc cagggagcac taagcgagca ctgcccaaca acaccagctc
    10021 ctctccccag ccaaagaaga aaccactgga tggagaatat ttcacccttc agatccgtgg
    10081 gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc ttggaactca aggatgccca
    10141 ggctgggaag gagccagggg ggagcagggc tcactccagc cacctgaagt ccaaaaaggg
    10201 tcagtctacc tcccgccata aaaaactcat gttcaagaca gaagggcctg actcagactg
    10261 accggtgttt aggcgcgccc ccctctccct cccccccccc taacgttact ggccgaagcc
    10321 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
    10381 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
    10441 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
    10501 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
    10561 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
    10621 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
    10681 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
    10741 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
    10801 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
    10861 acaaccatga gcttcgttgc tggagtcatc cggagactgg acgagacagt ggtgaacaga
    10921 attgccgccg gcgaggtgat ccagagacct gccaatgcaa taaaggagat gatcgagaac
    10981 tgtctggacg ccaagtccac aagcattcag gtgatcgtga aggagggcgg actgaagctg
    11041 atccagatcc aagacaacgg cacaggcatc agaaaggaag atctggacat cgtgtgtgaa
    11101 cggttcacca catctaagct gcagtctttt gaggatctgg cctctatcag tacctacggc
    11161 ttcagaggcg aggccctggc cagcatcagc cacgtggccc atgtgaccat caccaccaaa
    11221 accgccgacg gcaaatgcgc ttatcgcgct agctacagcg acggcaagct gaaagccccg
    11281 ccaaagcctt gcgccggcaa ccagggtaca cagataacag tggaggatct gttctacaac
    11341 atcgccaccc ggagaaaggc cctgaaaaat cccagcgagg agtacggcaa gatcctggaa
    11401 gtcgtcggca gatactccgt gcacaacgcc ggaatcagct ttagcgtaaa gaagcaggga
    11461 gaaaccgtgg ccgatgtgcg caccctgcca aatgccagca ccgtggataa catcagaagc
    11521 attttcggaa atgccgtgtc cagagaactg atcgagatcg gctgcgaaga taagaccctg
    11581 gcttttaaga tgaacggcta catcagcaac gccaattact ctgtgaagaa gtgcatcttt
    11641 cttctgttca tcaaccacag actggtggaa agcaccagcc tgcggaaagc catcgagaca
    11701 gtgtacgccg cctacctgcc taagaacacc caccccttcc tgtacctgag cctcgagatc
    11761 agccctcaga acgtggacgt caatgtgcat cctacaaagc acgaggtgca cttcctgcac
    11821 gaggaaagca tcctggaaag agtgcagcag cacattgaga gcaagctgct gggctctaac
    11881 agcagcagaa tgtacttcac acagaccctg ctgcctggcc tggccggccc ctcaggcgaa
    11941 atggttaagt ccacaacctc tctgacctca tccagcacca gcggttcttc cgataaggtg
    12001 tacgcccacc agatggtgcg gaccgactct cgggagcaga agctggacgc ctttctgcaa
    12061 cctctgagca aacctctgag ctctcagcct caggccatcg tgaccgagga caagacagat
    12121 atctcctccg gccgtgccag acagcaggac gaagaaatgc tcgagctgcc agctcctgcc
    12181 gaggtggccg ccaagaacca gagcctggag ggagatacca caaagggcac cagcgaaatg
    12241 agcgagaagc ggggccctac ctccagcaac cccagaaaac ggcaccggga ggacagcgac
    12301 gtggaaatgg tggaggacga cagcagaaag gaaatgacag ccgcttgtac ccctagaaga
    12361 agaatcatca acctgacctc cgtgctgagc ctgcaggagg agatcaacga gcagggccac
    12421 gaggtgctga gagagatgct gcacaatcac agcttcgtgg gctgcgtgaa ccctcaatgg
    12481 gccctggctc agcatcaaac aaagctgtac ctgctgaaca ccaccaagct gagcgaagag
    12541 ctgttctacc agatcctcat ctacgacttc gccaacttcg gcgtgctacg cctgagcgag
    12601 cccgcccctc tgtttgacct ggccatgctg gctctggata gcccagaaag cggctggaca
    12661 gaagaggacg gacctaaaga ggggctggct gaatacatcg tggagttcct gaagaaaaag
    12721 gccgagatgc tggccgacta cttttctctg gaaatcgacg aggaaggcaa cctgatcggc
    12781 ctgcctctgc tgatcgataa ctacgtgcct cccctggaag gcctgcccat cttcatcctg
    12841 agactggcta cagaggtgaa ctgggacgag gaaaaggaat gcttcgagtc tctgagcaag
    12901 gagtgcgcca tgttctatag catcagaaaa cagtacatct ctgaagagag cactctgtct
    12961 ggccagcaga gtgaagtgcc cggaagcatc cccaacagct ggaagtggac cgtggaacac
    13021 atcgtgtaca aggccctgcg gagccacatt ctccctccta agcacttcac cgaggacggc
    13081 aacatcctgc agctggccaa cctgcccgac ctttataagg ttttctaacg cgccgaccca
    13141 gctttcttgt acaaagtggt tgatccgggg atcctctagt cagcgtttat ctgcagaatt
    13201 cgcccttgtt taaacgatca taatcagcca taccacattt gtagaggttt tacttgcttt
    13261 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
    13321 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
    13381 aaataaagca tttttttcac tgcctcgacc catagagccc accgcatccc cagcatgcct
    13441 gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga
    13501 atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg
    13561 agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca
    13621 actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctctag actattcctt
    13681 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
    13741 atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
    13801 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
    13861 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg
    13921 cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc
    13981 caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc
    14041 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
    14101 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
    14161 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
    14221 cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc
    14281 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
    14341 catggcctcc gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa
    14401 cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctga attccccaat
    14461 gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc
    14521 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
    14581 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc
    14641 gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatctg
    14701 cagcctaggg ccgggattct cctccacgtc accgcatgtt agaagacttc ctctgccctc
    14761 tccgctgcca gatctctcga ggccctgtgg gaggaagaga agaggtcagg gatctgcccc
    14821 actgtggggt ggaggggaca gataaaagta cccagaacca gagccacatt aaccggccct
    14881 gggaatataa ggtggtccca gctcggggac acaggatccc tggaggcagc aaacatgctg
    14941 tcctgaagtg gacatagggg cccgggttgg aggaagaaga ctagctgagc tctcggaccc
    15001 ctggaagatg ccatgacagg gggctggaag agctagcaca gactagagag gtaagggggg
    15061 taggggagct gcccaaatga aaggagtgag aggtgacccg aatccacagg agaacggggt
    15121 gtccaggcaa agaaagcaag aggatggaga ggtggctaaa gccagggaga cggggtactt
    15181 tggggttgtc cagaaaaacg gtgatgatgc aggcctacaa gaaggggagg cgggacgcaa
    15241 gggagacatc cgtcggagaa ggccatccta agaaacgaga gatggcacag gccccagaag
    15301 gagaaggaaa agggaaccca gcgagtgaag acggcatggg gttgggtgag ggaggagaga
    15361 tgcccggaga ggacccagac acggggagga tccgctcaga ggacatcacg tggtgcagcg
    15421 ccgagaagga agtgctccgg aaagagcatc cttgggcagc aacacagcag agagcaaggg
    15481 gaagagggag tggaggaaga cggaacctga aggaggcggc agggaaggat ctgggccagc
    15541 cgtagaggtg acccaggcca caagctgcag acagaaagcg gcacaggccc aggggagaga
    15601 atgctggtca gagaaagcag gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
    15661 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
    15721 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
    15781 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
    15841 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
    15901 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
    15961 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
    16021 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
    16081 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
    16141 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
    16201 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
    16261 taacgcttac aatttag
//
