LOCUS       sequence-192699-       13735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of doxycycline-inducible constitutively active
            ROCK2 and co-expression of Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13735)
  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 13735)
  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 ROCK2
FEATURES             Location/Qualifiers
     source          1..13735
                     /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     complement(4580..4596)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4580..4596)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    complement(4627..4651)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4701..5912
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5749..5768
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(6037..6654,6655..6660,6661..6696,6697..6699,6700..6735,
                     6736..6738,6739..6774,6775..6783)
                     /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    6928..7390
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6984..7001)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7211..7230
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(7418..7420,7421..7423,7424..8137)
                     /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(7463..7484)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7724..7743)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8071..8092
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    8186..8774
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8239..8259)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8657..8668)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8777..8793)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8778..8794)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9303..9483
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9509..9526)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9515..9739
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9785..10213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9872..9891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10082..10103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10222..10242)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10227..10556
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10407..10542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        10604..10651
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10670..11044
                     /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    11174..11295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11211..11230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11265..11284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11344..11360)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11344..11360)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11357..11379)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11368..11384
                     /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(11392..11398,11399..11416,11417..11422))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11437..11458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11575..11592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11746..12334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11826..11845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12505..13296,13297..13365))
                     /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     13128..13147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13366..13470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13545..13564)
                     /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 gaattcctgc agcccggggg atccgggtac catgggccga cggggaagat
     2821 gcccggcgcc cccgaggccg tgtcggggga cggcgcgggc gcgagccgcc agaggaagct
     2881 ggaagcgctg atccgagacc ctcgttcgcc catcaacgtg gagagcttgc tggatggctt
     2941 aaattctttg gtccttgatt tggattttcc tgctttgagg aaaaacaaaa atatagataa
     3001 tttcttaaat agatatgaga aaattgtgaa aaaaattaga ggtttacaga tgaaggcaga
     3061 agactacgat gttgtaaaag ttatcggaag aggtgctttt ggtgaagtcc agttggttcg
     3121 tcataaggca tcacagaaag tttatgctat gaagcttctt agtaagtttg aaatgataaa
     3181 aagatcagat tctgcttttt tctgggagga aagagatatt atggcctttg ccaacagtcc
     3241 ctgggtggtt cagctctttt gtgcctttca agatgataag tatctgtaca tggtaatgga
     3301 gtacatgcct ggtggagacc ttgtaaacct tatgagtaac tatgatgtac ctgaaaaatg
     3361 ggccaaattt tatactgctg aagttgttct tgctttggat gccatacact ccatgggttt
     3421 aattcacaga gatgtgaagc ctgacaacat gctcttggat aaacatgggc atctaaaatt
     3481 agcagatttt ggcacatgta tgaagatgga tgaaacaggc atggtgcatt gtgatacagc
     3541 agttggaaca cccgattata tatcacccga ggtcctgaaa tcacaagggg gtgatggtta
     3601 ctatgggcga gaatgtgatt ggtggtccgt gggtgttttc ctttttgaaa tgctggtggg
     3661 ggatactcca ttttatgcag attcacttgt aggaacatat agcaaaatta tggatcataa
     3721 aaactcacta tgtttccctg aagatgcaga aatttctaaa catgcgaaga atctcatctg
     3781 tgccttctta acagataggg aggtacgcct tggaagaaac ggggtagaag aaatcaaaca
     3841 acatcctttc tttaagaatg atcagtggaa ttgggataac ataagagaga ctgcagctcc
     3901 tgtggtacct gaactcagca gtgacataga cagcagcaat tttgatgaca ttgaagatga
     3961 taaaggagat gtagaaacct tcccaattcc caaggctttt gtgggaaatc agctaccttt
     4021 tataggattt acctactaca gagaaaattt gctactaagt gactctccat cttgtaaaga
     4081 aaatgactca attcaatcaa ggaagaatga agagagtcaa gagattcaga aaaaactgta
     4141 cacattagaa gaacacctta gcactgagat tcaggccaaa gaggaactag aacagaagtg
     4201 caagtctgtt aatactcgct tagagaaagt ggcaaaggag ttagaagaag agattacctt
     4261 aaggaaaaat gtggaatcaa cattaagaca attagaaaga gaaaaagcac ttcttcagca
     4321 caaaaatgca gaatatcagc ggaaagctga tcatgaagca gacaagaagc gaaatttgga
     4381 gaatgatgtt aacagtttaa aagatcagct tgaagatttg aaaaaaagaa atcagaactc
     4441 tcagatatcc accctttcct gtatttaaaa caaaaaaaaa agtatttgcc agctcttagg
     4501 atgcaaattt gctttgcaga agaaaattag tgcactattt ttacacatag tagttatcat
     4561 tgtcggccca tggtacccgg atccactagt tctagaggcc gaggcggccg cactcgagat
     4621 atctagaccc agctttcttg tacaaagtgg ttgataacat ctcgagccat aaagatggtt
     4681 aattaaccca cccaagatct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     4741 cctcctcacg gcgagcgctg ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc
     4801 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     4861 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     4921 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     4981 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     5041 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     5101 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     5161 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     5221 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     5281 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     5341 acccaaggtc ttgaggcctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     5401 gggcaccatc tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg
     5461 tctgttgcgg gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga
     5521 gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg
     5581 gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc
     5641 tagggtaggc tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga
     5701 ggcgtcagtt tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt
     5761 tttgaactat gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg
     5821 aaatgtaatc atttgggtca atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta
     5881 aattctggcc gtttttggct tttttgttag acgaagcttg gtaccgagct cggatctcca
     5941 ccccgtaccg gtcctgcagg atttaaatgg cgcgccgcta gcgctaccgg actcagatct
     6001 cgagctcaag cttcgaattc tgcagtcgaa ttcaccatgt ctagactgga caagagcaaa
     6061 gtcataaacg gagctctgga attactcaat ggtgtcggta tcgaaggcct gacgacaagg
     6121 aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag
     6181 cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc
     6241 cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc ataccgctgt
     6301 gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa
     6361 cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag
     6421 aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa
     6481 caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca
     6541 cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc
     6601 ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg
     6661 accgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt
     6721 gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg
     6781 taactaagta aggatccgcg gccgcactag aggaattccg cccctctccc ccccccctaa
     6841 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     6901 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     6961 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     7021 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     7081 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     7141 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     7201 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     7261 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     7321 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     7381 acgatgataa gcttaccggt acgcgtgatg atccaccatg gtgagcaagg gcgaggagct
     7441 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     7501 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     7561 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     7621 cctgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     7681 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     7741 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     7801 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     7861 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     7921 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     7981 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     8041 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     8101 cgggatcact ctcggcatgg acgagctgta caagtaagaa ttgttgttgt ttagtccctc
     8161 ccaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     8221 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     8281 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     8341 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     8401 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     8461 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     8521 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     8581 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     8641 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     8701 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     8761 ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa catggagcaa
     8821 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     8881 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     8941 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     9001 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     9061 attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct
     9121 acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc
     9181 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt
     9241 ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta
     9301 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     9361 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     9421 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     9481 gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     9541 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     9601 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     9661 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     9721 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     9781 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     9841 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     9901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     9961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
    10021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
    10081 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
    10141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
    10201 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
    10261 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
    10321 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
    10381 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
    10441 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
    10501 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
    10561 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
    10621 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
    10681 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
    10741 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
    10801 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
    10861 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
    10921 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
    10981 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
    11041 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
    11101 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
    11161 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
    11221 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    11281 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
    11341 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
    11401 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
    11461 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
    11521 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
    11581 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
    11641 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
    11701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
    11761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
    11821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
    11881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
    11941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
    12001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
    12061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
    12121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
    12181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
    12241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
    12301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
    12361 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
    12421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
    12481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
    12541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
    12601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
    12661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
    12721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
    12781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
    12841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
    12901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
    12961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
    13021 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
    13081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    13141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    13201 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    13261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    13321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    13381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    13441 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
    13501 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
    13561 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
    13621 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
    13681 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
