LOCUS       sequence-159108-       14623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for doxycycline-inducible expression of FRNK in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14623)
  AUTHORS   Kim NG, Gumbiner BM
  TITLE     Adhesion to fibronectin regulates Hippo signaling via the 
            FAK-Src-PI3K pathway.
  JOURNAL   J Cell Biol. 2015 Aug 3;210(3):503-15. doi: 10.1083/jcb.201501025. 
            Epub 2015 Jul 27.
  PUBMED    26216901
REFERENCE   2  (bases 1 to 14623)
  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"; doi: 
            "10.1083/jcb.201501025"; journalName: "J Cell Biol"; date: 
            "2015-08-3- 3"; volume: "210"; issue: "3"; pages: "503-15"
COMMENT     Sequence Label: Tet-FRNK-GFP
FEATURES             Location/Qualifiers
     source          1..14623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /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      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2722..2740
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2793..2811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2829..2847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2864..2882
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2900..2918
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2930..2968
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2965..2989
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2998..3022
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4106..4130)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      4150..4159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4156..4158,4159..4161,4162..4875)
                     /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             join(4156..4158,4159..4161,4162..4872)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4201..4222)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4462..4481)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4809..4830
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           4926..5047
                     /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           5075..5089
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           5135..5247
                     /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(5342..5360)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5373..6584
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6421..6440
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    complement(6597..6630)
                     /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             6637..7344
                     /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(7345..7378)
                     /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    7401..7973
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7568..7585)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7795..7814
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7976..8578
                     /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     8475..8495
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8594..9182
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8647..8667)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9065..9076)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9389..9622
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9723..9740)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9729..9953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9999..10427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10086..10105)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10296..10317
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10436..10456)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10441..10770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10621..10756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10683..10702
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    11978..12099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12148..12164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12148..12164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12161..12183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12172..12188
                     /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(12196..12202,12203..12220,12221..12226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12241..12262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12379..12396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12550..13138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12630..12649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13309..14100,14101..14169))
                     /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     13932..13951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14170..14274)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    14322..14456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gacgagttta ctccctatca gtgatagaga acgatgtcga gtttactccc
     2761 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc
     2821 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat cagtgataga
     2881 gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag gtaggcgtgt
     2941 acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg caccggcaca
     3001 agtttgtaca aaaaagttgg catggaatcc aggcgacaag tcacagtatc ctgggactca
     3061 ggaggatcag atgaagctcc tcccaagccc agcaggcctg gttaccccag cccaaggtcc
     3121 agtgaagggt tttatccgag tcctcagcat atggtacagc caaatcacta ccaggtatct
     3181 ggctactctg gttctcatgg gataccagcc atggcaggca gcatttatcc tgggcaagct
     3241 tctctcttgg atcaaacaga ttcctggaac catcgacctc aggaagtatc agcatggcag
     3301 ccaaacatgg aggattcggg cactttggat gtacgaggaa tggggcaggt tctgcccaca
     3361 catctcatgg aggagaggtt aataagacaa cagcaagaga tggaagaaga tcaacgctgg
     3421 cttgagaaag aggaacgatt cctggtaatg aaacctgatg tgcggctctc cagaggcagc
     3481 attgaacggg aggacggagg tctccagggc ccagctggta accagcacat atatcagcct
     3541 gtgggtaaac cagatcatgc cgctccacca aagaagcccc ctcgccctgg agccccccac
     3601 ttgggcagcc tcgcgagcct gaacagcccc gtggacagct acaacgaagg cgtgaagatc
     3661 aagccacagg aaatcagccc tcctcctacg gccaacctgg accgctccaa tgacaaagtc
     3721 tatgagaatg taaccgggct ggtgaaagct gtcatagaga tgtccagtaa aatacagcca
     3781 gctccgccag aggagtacgt gcccatggta aaggaggttg gcttggcgct gagaaccttg
     3841 ctagcaacag tggatgagtc gctgccagtg cttcctgcaa gcacccacag agagattgag
     3901 atggcccaga aactgctgaa ctctgacctg gctgagctca ttaacaagat gaagctggcc
     3961 cagcagtacg tcatgaccag cctgcagcag gagtacaaga agcaaatgct gacggctgct
     4021 cacgctctgg ctgtggatgc caagaacttg ctggatgtca tcgatcaagc cagactgaaa
     4081 atgatcagcc agtccaggcc ccactaccca actttcttgt acaaagtggt tcgatgggat
     4141 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     4201 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     4261 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     4321 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     4381 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4441 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4501 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4561 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     4621 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     4681 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4741 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     4801 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4861 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatcgatag
     4921 tttgtttgaa tgaggcttca gtactttaca gaatcgttgc ctgcacatct tggaaacact
     4981 tgctgggatt acttcttcag gttaacccaa cagaaggctc gagaaggtat attgctgttg
     5041 acagtgagcg atctcgcttg ggcgagagta agtagtgaag ccacagatgt acttactctc
     5101 gcccaagcga gagtgcctcg gatcctgatc agaattcaag gggctacttt aggagcaatt
     5161 atcttgttta ctaaaactga ataccttgct atctctttga tacattttta caaagctgaa
     5221 ttaaaatggt ataaattaaa tcactttttt caattggaag actaatgcgg ccggccatta
     5281 ctccgtctcg tgtcttgttg catatgtctg ctggtttgtt tgatgttgtt tgcgggcggg
     5341 ccctatagtg agtcgtatta cctaggacgc gtggcctccg cgccgggttt tggcgcctcc
     5401 cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca gacgaagggc gcagcgagcg
     5461 tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag actcggcctt
     5521 agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact ctagggcact
     5581 ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg gcgattctgc
     5641 ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc cgggtgtggc
     5701 acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga
     5761 tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc cgccgggccg
     5821 ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg tccgcgagca
     5881 aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt tcccgagtct
     5941 tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg
     6001 tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg cgggaaagct cttattcggg
     6061 tgagatgggc tggggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     6121 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     6181 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     6241 atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc
     6301 agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg
     6361 agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc
     6421 tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt
     6481 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagactagta
     6541 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgctagc ggatccataa
     6601 cttcgtatag tatacattat acgaagttat gccaccatgt ctaggctgga caagagcaaa
     6661 gtcataaacg gagctctgga attactcaat ggtgtcggta tcgaaggcct gacgacaagg
     6721 aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag
     6781 cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc
     6841 cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc ataccgctgt
     6901 gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa
     6961 cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag
     7021 aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa
     7081 caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca
     7141 cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc
     7201 ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg
     7261 accgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgatttt
     7321 gaccttgaca tgctccccgg gtaaataact tcgtatagta tacattatac gaagttatgg
     7381 atccgcggcc gcaaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc
     7441 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     7501 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     7561 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     7621 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     7681 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     7741 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     7801 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     7861 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     7921 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatggc
     7981 caccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     8041 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     8101 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     8161 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     8221 cgtcgaagcg ggggcggtgt tcgccgagat cggctcgcgc atggccgagt tgagcggttc
     8281 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     8341 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     8401 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgct ggggtgcccg ccttcctgga
     8461 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     8521 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt
     8581 tcgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     8641 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     8701 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     8761 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     8821 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     8881 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     8941 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     9001 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     9061 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     9121 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     9181 gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac aagtagcaat
     9241 acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt
     9301 tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     9361 agccactttt taaaagaaaa gaggggactg gaagggctaa ttcactccca acgaagacaa
     9421 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9481 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9541 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9601 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     9661 cttgcaaaga aatgaatatc agagagtgag aggccttgac attgtttaaa cccgctgatc
     9721 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     9781 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     9841 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     9901 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     9961 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
    10021 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
    10081 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
    10141 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
    10201 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
    10261 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
    10321 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
    10381 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
    10441 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    10501 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
    10561 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
    10621 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
    10681 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
    10741 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
    10801 ggatctgatc agcacgtgat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt
    10861 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
    10921 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
    10981 gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt
    11041 ccggaagtgc ttgacattgg ggaattaatt cagcgagagc ctgacctatt gcatctcccg
    11101 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
    11161 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
    11221 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
    11281 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
    11341 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
    11401 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
    11461 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
    11521 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
    11581 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
    11641 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
    11701 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
    11761 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
    11821 cagcactcgt ccgagggcaa aggaatagca cgtgctacga gatttcgatt ccaccgccgc
    11881 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
    11941 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
    12001 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
    12061 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
    12121 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
    12181 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
    12241 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
    12301 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
    12361 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
    12421 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
    12481 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
    12541 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
    12601 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
    12661 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
    12721 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    12781 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    12841 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    12901 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    12961 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    13021 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    13081 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    13141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
    13201 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
    13261 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
    13321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
    13381 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
    13441 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
    13501 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
    13561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
    13621 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
    13681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
    13741 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
    13801 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
    13861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
    13921 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
    13981 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
    14041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
    14101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    14161 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
    14221 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
    14281 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt
    14341 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    14401 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
    14461 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
    14521 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
    14581 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
