LOCUS       sequence-375289-       13205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression vector for an inducible mCh-V5-NES-EGFP-SV40 
            NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13205)
  AUTHORS   Wang L, Paudel BB, McKnight RA, Janes KA
  TITLE     Nucleocytoplasmic transport of active HER2 causes fractional escape 
            from the DCIS-like state.
  JOURNAL   Nat Commun. 2023 Apr 13;14(1):2110. doi: 10.1038/s41467-023-37914-x.
  PUBMED    37055441
REFERENCE   2  (bases 1 to 13205)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2023-04-13"; volume: "14(1):2110. doi"; pages: " 
            10.1038/s41467-023-37914-x"
COMMENT     Sequence Label: pSLIK mCh-V5-NES-EGFP-SV40 NLS zeo
FEATURES             Location/Qualifiers
     source          1..13205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2614..2638
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        2670..2984
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2678..2696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2714..2732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2749..2767
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2785..2803
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2821..2839
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2856..2874
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2892..2910
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2957..2981
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2991..3698
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3136..3154)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3404..3423
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3699..3740
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             join(3777..3779,3780..3782,3783..4490)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(3822..3843)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4083..4102)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4430..4451
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4497..4517
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(4556..4580)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4630..5841
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5678..5697
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5900..6517,6518..6523,6524..6559,6560..6562,6563..6598,
                     6599..6601,6602..6637,6638..6646)
                     /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    6684..7257
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6851..6868)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7078..7097
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     promoter        7279..7326
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7345..7719
                     /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"
     misc_feature    7893..8481
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7946..7966)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8364..8375)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8484..8500)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8485..8501)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9010..9190
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9216..9233)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9222..9446
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9492..9920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9579..9598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9789..9810
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9929..9949)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9934..10263
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10114..10249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10176..10195
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10311..10358
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10377..10751
                     /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    10881..11002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10918..10937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10972..10991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11051..11067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11051..11067)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11064..11086)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11075..11091
                     /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(11099..11105,11106..11123,11124..11129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11144..11165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11282..11299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11453..12041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11533..11552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12212..13003,13004..13072))
                     /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     12835..12854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13073..13177)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaattc tagacagtta tcaacaagtt tgtacaaaaa agcaggcttt
     2641 aaaggaacca attcagtcga ctggatcccg agtttactcc ctatcagtga tagagaacgt
     2701 atgtcgagtt tactccctat cagtgataga gaacgatgtc gagtttactc cctatcagtg
     2761 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
     2821 tccctatcag tgatagagaa cgtatgtcga gtttatccct atcagtgata gagaacgtat
     2881 gtcgagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg
     2941 aggcctatat aagcagagct cgtttagtga accgtcagat cgccactagt atggtgagca
     3001 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
     3061 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     3121 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
     3181 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
     3241 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     3301 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca
     3361 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
     3421 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
     3481 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
     3541 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
     3601 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg
     3661 gccgccactc caccggcggc atggacgagc tgtacaaggg taagcctatc cctaaccctc
     3721 tcctcggtct cgattctacg accggtttag ccttgaaatt agcaggtctt gatatcatgg
     3781 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     3841 acgtaaacgg ccacaagttc agcgtgtctg gcgagggcga gggcgatgcc acctacggca
     3901 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     3961 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     4021 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     4081 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     4141 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     4201 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     4261 tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     4321 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     4381 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     4441 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac ggtaccccaa
     4501 aaaagaagag aaaggtatga caattggaag actaatgcgt tttcgagata tctagaccca
     4561 gctttcttgt acaaagtggt tgataacatc tcgagccata aagatggtta attaacccac
     4621 ccaagatctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg
     4681 cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc
     4741 tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg
     4801 acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa
     4861 caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg
     4921 aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg
     4981 gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc
     5041 tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg
     5101 agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg
     5161 gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc
     5221 gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct
     5281 tgaggccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg ggcaccatct
     5341 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     5401 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     5461 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     5521 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     5581 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     5641 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     5701 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     5761 tttgggtcaa tatgtaattt tcagtgttag actagtaaat tgtccgctaa attctggccg
     5821 tttttggctt ttttgttaga cgaagcttgg taccgagctc ggatctccac cccgtaccgg
     5881 tcctgcagtc gaattcacca tgtctagact ggacaagagc aaagtcataa acggagctct
     5941 ggaattactc aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct
     6001 gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc
     6061 cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc
     6121 atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg
     6181 cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga
     6241 aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct
     6301 gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc
     6361 aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat
     6421 tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat
     6481 atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga
     6541 ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc
     6601 tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcc
     6661 gcggccgcac tagaggaatt ccgcccctct cccycccccc cccctaacgt tactggccga
     6721 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     6781 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     6841 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     6901 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     6961 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     7021 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     7081 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     7141 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     7201 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagct
     7261 taccggtacg cgtgatgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     7321 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     7381 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     7441 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     7501 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     7561 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     7621 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     7681 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acatctgtac aagtaaagcg
     7741 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     7801 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttta
     7861 gtccctccca attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     7921 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     7981 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     8041 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     8101 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     8161 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     8221 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     8281 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     8341 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     8401 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     8461 ccctttgggc cgcctccccg catcgatacc gtcgacctcg atcgagacct agaaaaacat
     8521 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     8581 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     8641 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     8701 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     8761 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     8821 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     8881 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     8941 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     9001 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     9061 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     9121 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     9181 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     9241 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     9301 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     9361 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     9421 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     9481 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     9541 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     9601 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     9661 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     9721 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     9781 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     9841 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     9901 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     9961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
    10021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    10081 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
    10141 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
    10201 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
    10261 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
    10321 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
    10381 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
    10441 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
    10501 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
    10561 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
    10621 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
    10681 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
    10741 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
    10801 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
    10861 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
    10921 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
    10981 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
    11041 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
    11101 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
    11161 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
    11221 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
    11281 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
    11341 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
    11401 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
    11461 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
    11521 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
    11581 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
    11641 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
    11701 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
    11761 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
    11821 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
    11881 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
    11941 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
    12001 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    12061 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    12121 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    12181 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    12241 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    12301 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    12361 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    12421 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    12481 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    12541 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    12601 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    12661 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    12721 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    12781 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    12841 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    12901 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    12961 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    13021 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    13081 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    13141 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    13201 ctgac
//
