LOCUS       sequence-204053-       14724 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for Tet-inducible WTIP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14724)
  AUTHORS   Ibar C, Kirichenko E, Keepers B, Enners E, Fleisch K, Irvine KD
  TITLE     Tension-dependent regulation of mammalian Hippo signaling through 
            LIMD1.
  JOURNAL   J Cell Sci. 2018 Feb 13. pii: jcs.214700. doi: 10.1242/jcs.214700.
  PUBMED    29440237
REFERENCE   2  (bases 1 to 14724)
  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: "J Cell Sci.
            2018 Feb 13. pii: jcs.214700. doi: 10.1242/jcs.214700."
COMMENT     Sequence Label: pTRIPZ-EGFP:WTIP
FEATURES             Location/Qualifiers
     source          1..14724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    668..789
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(838..854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(838..854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(851..873)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    862..878
                     /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(886..892,893..910,911..916))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    931..952
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1069..1086)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1240..1828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1320..1339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1999..2790,2791..2859))
                     /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     2622..2641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2860..2964)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    3012..3146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             3314..3947
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3994..4119
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4616..4849
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5034..5078
                     /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    5376..5493
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5558..5932)
                     /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"
     regulatory      5929..5938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5942..5960)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5952..5999)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    6035..6053
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6070..6088
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6106..6124
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6142..6160
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6177..6195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6213..6231
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        6243..6281
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6278..6302
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(6306..6308,6309..6311,6312..7022)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6306..6308,6309..6311,6312..7022)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6351..6372)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6612..6631)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6959..6980
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           8430..8523
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           8549..8661
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        complement(8756..8774)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        8787..9998
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     9835..9854
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(10011..10044)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     CDS             10051..10758
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     protein_bind    complement(10759..10792)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     misc_feature    10815..11387
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(10982..10999)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     11209..11228
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             11390..11992
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     11889..11909
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    12008..12596
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12061..12081)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12479..12490)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12803..13036
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(13137..13154)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13143..13367
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      13413..13841
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13500..13519)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13710..13731
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(13850..13870)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13855..14184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      14035..14170
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     14097..14116
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
       61 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
      121 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
      181 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
      241 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
      301 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
      361 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
      421 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
      481 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
      541 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
      601 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
      661 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
      721 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
      781 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
      841 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      901 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
      961 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1021 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1081 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1141 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1201 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1261 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1321 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1381 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1441 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1501 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1561 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1621 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1681 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1741 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1801 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1861 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1921 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1981 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2041 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2101 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2161 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2221 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2281 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2341 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2401 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2461 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2521 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2581 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2641 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2701 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2761 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2821 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     2881 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     2941 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     3001 atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3061 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3121 tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca
     3181 aacttcccac cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa
     3241 cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta
     3301 caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat ccttgatctg
     3361 tggatctacc acacacaagg ctacttccct gattagcaga actacacacc agggccaggg
     3421 gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gccagataag
     3481 gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg
     3541 atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat
     3601 cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac
     3661 aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg
     3721 cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt
     3781 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     3841 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     3901 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     3961 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     4021 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     4081 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     4141 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     4201 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     4261 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     4321 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     4381 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     4441 accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg
     4501 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     4561 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     4621 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     4681 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     4741 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     4801 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     4861 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     4921 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     4981 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     5041 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     5101 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     5161 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     5221 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     5281 gtggagagag agacagagac agatccattc gattagtgaa cggatcggca ctgcgtgcgc
     5341 caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     5401 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     5461 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     5521 ccagtttggt tagtaccggg cccgctctag tccggaatca gtcctgctcc tcggccacga
     5581 agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc
     5641 cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg acctccgacc
     5701 actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg ttgtccggca
     5761 ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc acaccggcga
     5821 agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac tcgaccgctc
     5881 cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc atggtggccc
     5941 tcctatagtg agtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca
     6001 cgtgctgcag gtccgaggtt ctagacgagt ttactcccta tcagtgatag agaacgatgt
     6061 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
     6121 gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttatccc
     6181 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
     6241 gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg aaccgtcaga
     6301 tcgcaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     6361 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     6421 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     6481 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     6541 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     6601 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     6661 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     6721 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     6781 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     6841 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     6901 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     6961 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     7021 agtactcaga tctcgagctc aagcttcgaa ttctgcagtc gacggtacca atgcagcgct
     7081 ccagggcggg cgcggacgag gcggccctac tcctggccgg gctggccctg cgggagctgg
     7141 agcccgggtg cggctctccc ggtcgggggc ggcgggggcc gcggcctggg cctggagacg
     7201 aggcggcgcc cgcgctgggc cgcagaggga agggcagcgg cggccccgag gccggggcgg
     7261 acggactgag ccgcggggag cggggtcccc ggcgcgcggc ggttccggag ctcagcgcgc
     7321 agcctgcggg cagcccacgg gccagcctgg cggggtccga cggcggcggc ggtggcggca
     7381 gcgcccgatc cagcggcatc agcctgggct acgaccagcg ccacggcagc ccgcgctccg
     7441 gtcgctcgga cccgcgtccc ggtcccgggc cgccttcggt gggcagcgcc cgctccagcg
     7501 tttccagcct cggctcccgg ggctcggccg gcgcctacgc tgacttcctc ccgcccggcg
     7561 cctgccccgc gcccgctcgc tccccggagc ctgcggggcc ggctcccttc ccgctgcctg
     7621 cactcccgct gccccctggc cgggagggcg gcccaagcgc ggccgagcgg cggctggagg
     7681 cgctcacccg ggagctggag cgggcgctcg aggcgcgcac ggcgcgggac tacttcggca
     7741 tttgcatcaa gtgtgggctt ggcatctacg gagcccagca ggcgtgccag gcaatgggga
     7801 gtctttatca cactgactgc ttcacctgcg actcgtgtgg gagacgactc cgtgggaagg
     7861 cgttctacaa cgtgggtgag aaagtgtact gccaggagga cttcctgtac tccgggttcc
     7921 agcagacggc cgacaaatgc agcgtgtgtg gacatctcat catggaaatg atcctgcagg
     7981 ccctgggcaa gtcctaccac ccaggctgct tccggtgctc cgtgtgcaat gagtgcctgg
     8041 acggggttcc cttcaccgtg gacgtggaga acaacatcta ctgcgtgcga gactatcaca
     8101 cggtttttgc accaaaatgc gcctcctgtg cccgtcctat cctccctgca cagggctgcg
     8161 agacaaccat ccgtgtggtg tccatggaca gagactacca cgtggcatgt taccactgtg
     8221 aggactgcgg gctgcagctg agcggggagg agggacgccg ttgctatccc ctggcgggcc
     8281 acctactgtg tcgtcgttgc cacctgcggc gcctccaacc tgggcctctt ccctcaccca
     8341 ctgtgcacgt cactgagctc tgatctagat aactgatcat aatcagccat accacatttg
     8401 tagaggtttt acttgcttta atagtttgtt tgaatgaggc ttcagtactt tacagaatcg
     8461 ttgcctgcac atcttggaaa cacttgctgg gattacttct tcaggttaac ccaacagaag
     8521 gctcgaggta accggatcct gatcagaatt caaggggcta ctttaggagc aattatcttg
     8581 tttactaaaa ctgaatacct tgctatctct ttgatacatt tttacaaagc tgaattaaaa
     8641 tggtataaat taaatcactt ttttcaattg gaagactaat gcggccggcc attactccgt
     8701 ctcgtgtctt gttgcatatg tctgctggtt tgtttgatgt tgtttgcggg cgggccctat
     8761 agtgagtcgt attacctagg acgcgtggcc tccgcgccgg gttttggcgc ctcccgcggg
     8821 cgcccccctc ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg agcgtcctga
     8881 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     8941 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     9001 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     9061 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     9121 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     9181 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     9241 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     9301 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     9361 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     9421 gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa agctcttatt cgggtgagat
     9481 gggctggggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
     9541 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
     9601 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
     9661 ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
     9721 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
     9781 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
     9841 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
     9901 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact agtaaattgt
     9961 ccgctaaatt ctggccgttt ttggcttttt tgttagacgc tagcggatcc ataacttcgt
    10021 atagtataca ttatacgaag ttatgccacc atgtctaggc tggacaagag caaagtcata
    10081 aacggagctc tggaattact caatggtgtc ggtatcgaag gcctgacgac aaggaaactc
    10141 gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc
    10201 ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg
    10261 gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc
    10321 ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac
    10381 gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca
    10441 ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag
    10501 catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg
    10561 agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg
    10621 gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac
    10681 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ttttgacctt
    10741 gacatgctcc ccgggtaaat aacttcgtat agtatacatt atacgaagtt atggatccgc
    10801 ggccgcaaat tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg
    10861 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
    10921 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
    10981 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
    11041 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
    11101 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
    11161 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
    11221 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
    11281 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc
    11341 cccccgaacc acggggacgt ggttttcctt tgaaaaacac gataatacca tggccaccga
    11401 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
    11461 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
    11521 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
    11581 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
    11641 agcgggggcg gtgttcgccg agatcggctc gcgcatggcc gagttgagcg gttcccggct
    11701 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
    11761 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
    11821 cgtgctcccc ggagtggagg cggccgagcg cgctggggtg cccgccttcc tggagacctc
    11881 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
    11941 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagttcgcgt
    12001 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
    12061 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
    12121 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
    12181 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
    12241 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
    12301 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
    12361 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
    12421 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
    12481 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
    12541 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
    12601 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
    12661 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
    12721 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
    12781 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatctg
    12841 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
    12901 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
    12961 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
    13021 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
    13081 aagaaatgaa tatcagagag tgagaggcct tgacattgtt taaacccgct gatcagcctc
    13141 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
    13201 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
    13261 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
    13321 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
    13381 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
    13441 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
    13501 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
    13561 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
    13621 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
    13681 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
    13741 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
    13801 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
    13861 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    13921 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
    13981 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
    14041 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
    14101 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
    14161 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
    14221 gatcagcacg tgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
    14281 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
    14341 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
    14401 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
    14461 gtgcttgaca ttggggaatt aattcagcga gagcctgacc tattgcatct cccgccgtgc
    14521 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
    14581 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
    14641 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
    14701 tgatccccat gtgtatcact ggca
//
