LOCUS       sequence-257022-       15904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of doxycycline-inducible constitutively active
            MLCK and co-expression of Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15904)
  AUTHORS   Wong SY, Ulrich TA, Deleyrolle LP, MacKay JL, Lin JM, Martuscello 
            RT, Jundi MA, Reynolds BA, Kumar S
  TITLE     Constitutive activation of myosin-dependent contractility sensitizes
            glioma tumor-initiating cells to mechanical inputs and reduces 
            tissue invasion.
  JOURNAL   Cancer Res. 2015 Mar 15;75(6):1113-22. doi: 
            10.1158/0008-5472.CAN-13-3426. Epub 2015 Jan 29.
  PUBMED    25634210
REFERENCE   2  (bases 1 to 15904)
  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: 
            "10.1158/0008-5472.CAN-13-3426"; journalName: "Cancer Res"; date: 
            "2015-03-15- 15"; volume: "75"; issue: "6"; pages: "1113-22"
COMMENT     Sequence Label: pSLIK CA MLCK
FEATURES             Location/Qualifiers
     source          1..15904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2377..2401
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        2433..2747
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2441..2459
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2477..2495
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2512..2530
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2548..2566
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2584..2602
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2619..2637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2655..2673
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2720..2744
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     2807..2823
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6466..6489
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(6780..6804)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        6854..8064
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7901..7920
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(8189..8806,8807..8812,8813..8848,8849..8851,8852..8887,
                     8888..8890,8891..8926,8927..8935)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    8986..9559
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9153..9170)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9380..9399
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(9587..9589,9590..9592,9593..10306)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9632..9653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9893..9912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10240..10261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    10355..10943
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10408..10428)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10826..10837)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10946..10962)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             11472..11652
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11678..11695)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11684..11908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11954..12382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12041..12060)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12251..12272
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12391..12411)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12396..12725
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12576..12711
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12638..12657
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12773..12820
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12839..13213
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    13343..13464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13380..13399)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13434..13453
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13513..13529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13513..13529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13526..13548)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13537..13553
                     /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(13561..13567,13568..13585,13586..13591))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13606..13627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13744..13761)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13915..14503)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13995..14014)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14674..15465,15466..15534))
                     /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     15297..15316
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15535..15639)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(15714..15733)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttctagacag ttatcaacaa gtttgtacaa aaaagcaggc
     2401 tttaaaggaa ccaattcagt cgactggatc ccgagtttac tccctatcag tgatagagaa
     2461 cgtatgtcga gtttactccc tatcagtgat agagaacgat gtcgagttta ctccctatca
     2521 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt
     2581 tactccctat cagtgataga gaacgtatgt cgagtttatc cctatcagtg atagagaacg
     2641 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgaggtagg cgtgtacggt
     2701 gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgccact agtcgactgg
     2761 atccggtacc ggccgcctgg gccggtgacc gtaccgggcc ccccctcgag gtcgacggta
     2821 tcgataagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt gctggaattc
     2881 tgcagatatc catcacactg gcggccgcag gcgatccgcc agcaggaggt ggagcagctg
     2941 gacttccgtg agctgctggg gaagaaggtg agtaccaaga ccctgtcgga ggaagccctg
     3001 aaggagatcc cggccgagca gatggatttc cgcgccaacc tgcagcggca ggtgaagccc
     3061 aagactgtgt ctgaggagga gaggaaggtg cacagccccc agcaggtcga cttccgctcc
     3121 gtcctagcca agaaggggac tcccaagacc cccgtgcctg agaaggcgcc acctccaaaa
     3181 cctgccactc cggattttcg ctccgtgctg ggtagcaaga agaaattacc ggcagagaat
     3241 ggtagcagca atgctgaggc cttgaatgtc aaggcaacag aaagccccaa gcttgtgggc
     3301 aatgcaccac tgtcagggtc cctgaaacct gtggccaacg ccaagcctgc agagaccctg
     3361 aaacctgtgg ccaacaccaa acctgccgag accctgaaac ctgtggccaa cgcagagacc
     3421 ctgaaaccca tgggcaacgc caaacctgcc gagagctcga aacctgtggg caacaccaaa
     3481 cctgccgaga ccctgaaacc cgtgggcaac atcaaacctg ccgagaccct gaaacccgtg
     3541 ggcaacatca aacctgccga gaccctgaaa cccgtgggca acatcaaacc tgccgagacc
     3601 ttgaaacccg tgggcaacac caagcctact gagaccctga aacctgtggc caatgccaag
     3661 cctgccgaga ccctgaaacc cgtgggcaac accaagcctg ccgagaccct gaaacccgtg
     3721 ggcaatgcca agcctgctga gaccctgaaa cccgtgggca acgccaagcc tgctgagacc
     3781 ctgaaacccg tgggcaatgc caagcctgcc gagaccctga aacctgtggg caacgccaag
     3841 cctgccgaga ccctgaaagc cgtggccaac gcaaagcctg ccgagacccc caaaccagca
     3901 gggaaagagg aactcaagaa ggaggtccag aatgatgtaa actgcaagag agagaaggca
     3961 ggagctgcgg ataacgaaaa accacctgcg agcccaggta cagccccaac cttcaaggag
     4021 aagctgcagg acgtccgtgt ggcagagggt gagaagctgc tgctccagtg ccaggtgtcc
     4081 tctgagcccc cggccaccat cacctggaca ctgaacggga agacactcaa gaccaccaaa
     4141 ttcgtcatcc tctcccaaga aggctcgctg tgctccgtct ccattgagaa ggcgctgccg
     4201 gaggacagag ggctgtacaa gtgtgtggcc aagaatgccg caggccaggc cgagtgctcc
     4261 tgccacgtca ccgtgcacga tgccccggcc agcgagaatg ccaaggcccc ggagatgaaa
     4321 tcccggaggc ccaagagctc tcttcccccc gtgctaggaa cagagagtga cgccactgtg
     4381 aaaaagaaac ctgcccccaa gacgcccccg aaagcagcca tgccccctca gatcctccag
     4441 tttcctgagg accaaaaggt gcgcgcggga gagcgtgtgg agctgtttgg gaaggtggcc
     4501 ggcacccagc ccatcacctg tacctggatg aagttccgca agcagatcca ggacagcgag
     4561 cacatcaagg tggagaacag cgaggctggc agcaagctca ccatcctggc cgcacgccag
     4621 gagcattgtg gctgctacac gctgctggtg gagaacaagc tgggcagcag gcaggcccag
     4681 gttaacctca ctgtcgtgga taagccagac ccccccgccg gcacgccgtg cgcctctgac
     4741 atccgcagct cctcgctgac gctgtcctgg tacggatcct cctacgacgg gggcagcgcc
     4801 gtgcagtcct acagcgtgga gatctgggac tctgtggaca agatgtggac ggagctggcc
     4861 acatgccgca gcacctcgtt caacgtgcgc gacctgctgc cggaccgcga atataaattc
     4921 cgcgtgcgcg caatcaacgt gtacggcacc agtgagccga gccaggagtc ggagctcacg
     4981 acagtgggcg agaagcccga ggagccgaag gatgaagtgg aggaggtgtc ggatgatgac
     5041 gagaaggagc ccgaggtcga ctaccggaca gtgaccgtca acaccgagca gaaagtgtct
     5101 gacttctatg acatcgaaga gaggctagga tctgggaaat ttggacaggt cttccgactc
     5161 gtagagaaga aaactggaaa aatctgggca gggaagttct tcaaggcgta ctcggccaaa
     5221 gagaaggaga acatccggca ggagattggc atcatgaact gcctccacca ccccaagctg
     5281 gtgcagtgcg tggacgcctt cgaggaaaag gccaacatcg tcatggtcct tgagatagta
     5341 tcaggtggcg agctgtttga gcgcatcatc gacaagaagt tcgagctgac ggagcgcgag
     5401 tgcatcaagt acatgcgtca gatctcggag ggggtggagt acatccacaa gcagggcatc
     5461 gtccacctcg acctcaagcc tgagaacatc atgtgcgtca acaagacggg caccaggatc
     5521 aagctcatcg acttcggcct ggccagaagg ctggagaacg ccgggtctct gaaggtcctc
     5581 ttcggcaccc cggaattcgt ggcacccgaa gtgatcaact acgagcccat cagctatgcc
     5641 acggacatgt ggagcatcgg cgtcatctgc tacatcctgg tcagcggcct ctcccccttc
     5701 atgggtgaca atgacaatga gaccttagcc aatgttacct cagccacctg ggacttcgac
     5761 gacgaggcat ttgatgagat ctccgacgac gccaaggatt tcatcagcaa cttgctgaag
     5821 aaggacatga aaaaccgcct ggactgtacc cagtgcctcc agcacccgtg gctaatgaag
     5881 gataccaaga acatggaggc caagaagctc tccaaggacc gcatgaagaa gtacatggca
     5941 agaagaaaat ggcagaaaac gggcaatgct gtgagagcca ttggaagact gtcctctatg
     6001 gcaatgatct cagggctcag cggcaggaaa tcctccacgg ggtcacccac cagcccactc
     6061 accgcagaga gactggaaac cgaagaagat gtctcccaag cgttcctgga ggctgtggca
     6121 gaggagaagc cgcacgtgaa accttacttc tctaagacca ttcgtgacct ggaggtggtg
     6181 gagggcagcg ccgccagatt tgactgcaag atcgaaggat acccagaccc cgaggtcgtc
     6241 tggttcaaag acgaccagtc aatcagggag tctcgccact tccagataga ctacgacgaa
     6301 gacgggaact gctctttaat tatcagcgac gtgtgcgggg acgacgatgc caagtacacc
     6361 tgcaaggctg tcaacagtct tggagaagcc acctgcacgg cagagctcat tgtggaaacc
     6421 atggaggaag gagaaggaga aggggaagag gaggaagagg aggttgacta caaggacgac
     6481 gatgacaagt gaaacaaagc cagggggaag cagtttgtaa gtcgtgttac aaggactaac
     6541 tctctaaagc tcagaaactc aagatagcaa aagcacctag tgtggcaggg tctctaggta
     6601 ggtcccttgg gggctgatgg ttcgccacag cagttgaaac tcagcagcat cgctgatggg
     6661 cattaatctg aattagctgg caccttaaga tcctggtgca gcaggatttc atttagggaa
     6721 atttccagta acttgcctgg tgaattccag tctagagcgg ccgcactcga gatatctaga
     6781 cccagctttc ttgtacaaag tggttgataa catctcgagc cataaagatg gttaattaac
     6841 ccacccaaga tctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc
     6901 acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc gtcctgatcc tccgcccgga
     6961 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     7021 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     7081 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     7141 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     7201 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     7261 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     7321 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     7381 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     7441 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     7501 tcttgaggcc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tggggcacca
     7561 tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc
     7621 gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc
     7681 cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccacctg
     7741 ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag
     7801 gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag
     7861 tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact
     7921 atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa
     7981 tcatttgggt caatatgtaa ttttcagtgt tagactagta aattgtccgc taaattctgg
     8041 ccgtttttgg cttttttgtt agacgaagct tggtaccgag ctcggatctc caccccgtac
     8101 cggtcctgca ggatttaaat ggcgcgccgc tagcgctacc ggactcagat ctcgagctca
     8161 agcttcgaat tctgcagtcg aattcaccat gtctagactg gacaagagca aagtcataaa
     8221 cggagctctg gaattactca atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc
     8281 tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct
     8341 gctcgatgcc ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga
     8401 aggcgagtca tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct
     8461 ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga
     8521 aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact
     8581 gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca
     8641 tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag
     8701 acaagcaatt gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga
     8761 actaatcata tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc
     8821 ccttgacgat tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga
     8881 tatgctgcct gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaactaag
     8941 taaggatccg cggccgcact agaggaattc cgcccctccc ccccccccct ctccctcccc
     9001 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     9061 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     9121 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     9181 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     9241 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     9301 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     9361 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     9421 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     9481 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     9541 ttgaaaaaca cgatgataag cttaccggta cgcgtgatga tccaccatgg tgagcaaggg
     9601 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     9661 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     9721 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     9781 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     9841 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     9901 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     9961 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
    10021 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
    10081 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
    10141 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
    10201 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
    10261 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat tgttgttgtt
    10321 tagtccctcc caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
    10381 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
    10441 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
    10501 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
    10561 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
    10621 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
    10681 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
    10741 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
    10801 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
    10861 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
    10921 ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac
    10981 atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag
    11041 cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga
    11101 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
    11161 taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct
    11221 acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg
    11281 gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag
    11341 agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
    11401 tattagagtg gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc
    11461 cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
    11521 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
    11581 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
    11641 aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
    11701 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
    11761 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
    11821 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
    11881 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
    11941 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
    12001 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
    12061 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
    12121 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
    12181 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
    12241 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
    12301 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
    12361 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
    12421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
    12481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
    12541 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
    12601 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
    12661 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
    12721 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
    12781 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
    12841 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
    12901 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
    12961 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa
    13021 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
    13081 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
    13141 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
    13201 ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
    13261 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
    13321 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
    13381 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
    13441 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
    13501 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
    13561 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
    13621 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
    13681 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
    13741 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
    13801 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
    13861 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
    13921 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
    13981 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
    14041 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
    14101 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
    14161 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
    14221 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
    14281 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
    14341 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
    14401 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
    14461 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
    14521 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
    14581 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
    14641 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
    14701 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
    14761 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
    14821 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
    14881 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
    14941 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
    15001 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
    15061 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
    15121 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
    15181 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
    15241 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
    15301 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
    15361 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
    15421 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
    15481 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
    15541 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
    15601 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
    15661 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
    15721 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
    15781 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
    15841 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
    15901 cgtt
//
