LOCUS       sequence-12070-e       13217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control lentiviral vector with Tet-based inducible expression of 
            Luciferase miR30-based shRNA and GFP, constitutive Neomycin 
            resistance gene coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13217)
  AUTHORS   Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI, 
            Rebres R, Roach T, Seaman W, Simon MI, Fraser ID
  TITLE     A single lentiviral vector platform for microRNA-based conditional 
            RNA interference and coordinated transgene expression.
  JOURNAL   Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.
  PUBMED    16945906
REFERENCE   2  (bases 1 to 13217)
  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: "Proc Natl 
            Acad Sci U S A. 2006 Sep 12. 103(37):13759-64."
COMMENT     Sequence Label: pSLIK-Neo-TGmiR-Luc
FEATURES             Location/Qualifiers
     source          1..13217
                     /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    2610..2634
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    join(2673..2691,2692..2714,2715..2733,2734..2756,2757..2775,
                     2776..2798,2799..2817,2818..2840,2841..2859,2860..2882,
                     2883..2901,2902..2924,2925..2943)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2976..3014
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3011..3035
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(3121..3123,3124..3126,3127..3840)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3121..3123,3124..3126,3127..3837)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3166..3187)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3427..3446)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3774..3795
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           3872..3993
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4021..4035
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4063..4189
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(4228..4252)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4299..5510
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5347..5366
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5569..6186,6187..6192,6193..6228,6229..6231,6232..6267,
                     6268..6270,6271..6306,6307..6315)
                     /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    6353..6927
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6520..6537)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             6943..7737
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6943..7737
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6997..7016)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7607..7626
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    7905..8493
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7958..7978)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8376..8387)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8496..8512)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8497..8513)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9022..9202
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9228..9245)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9234..9458
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9504..9932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9591..9610)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9801..9822
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9941..9961)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9946..10275
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10126..10261
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10188..10207
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10323..10370
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10389..10763
                     /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    10893..11014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10930..10949)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10984..11003
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11063..11079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11063..11079)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11076..11098)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11087..11103
                     /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(11111..11117,11118..11135,11136..11141))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11156..11177
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11294..11311)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11465..12053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11545..11564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12224..13015,13016..13084))
                     /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     12847..12866
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13085..13189)
                     /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 caagtttgta caaaaaagca ggctttaaag
     2641 gaaccaattc agtcgactgg atcctttacc actccctatc agtgatagag aaaagtgaaa
     2701 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
     2761 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
     2821 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     2881 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     2941 agaaaagtga aagtcgagct cggtacccgg gtcgaggtag gcgtgtacgg tgggaggcct
     3001 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
     3061 ttgacctcca tagaagacac cgggaccgat ccagcctccg cggccccgaa ctagtccacc
     3121 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3181 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3241 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3301 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3361 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3421 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3481 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3541 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3601 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3661 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3721 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3781 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     3841 actagtcgac tagggataac agggtaattg tttgaatgag gcttcagtac tttacagaat
     3901 cgttgcctgc acatcttgga aacacttgct gggattactt cttcaggtta acccaacaga
     3961 aggctcgaga aggtatattg ctgttgacag tgagcgcccg cctgaagtct ctgattaata
     4021 gtgaagccac agatgtatta atcagagact tcaggcggtt gcctactgcc tcggaattca
     4081 aggggctact ttaggagcaa ttatcttgtt tactaaaact gaataccttg ctatctcttt
     4141 gatacatttt tacaaagctg aattaaaatg gtataaatta aatcactttt ttcaattgga
     4201 agactaatgc gttttcgaga tatctagacc cagctttctt gtacaaagtg gttaacatct
     4261 cgagccataa agatggttaa ttaacccacc caagatctgg cctccgcgcc gggttttggc
     4321 gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag
     4381 cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     4441 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     4501 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     4561 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     4621 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
     4681 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     4741 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
     4801 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
     4861 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     4921 gcatggtggg cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta
     4981 ttcgggtgag atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     5041 ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt
     5101 gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg
     5161 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
     5221 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     5281 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     5341 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     5401 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     5461 ctagtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac gaagcttggt
     5521 accgagctcg gatctccacc ccgtaccggt cctgcagtcg aattcaccat gtctagactg
     5581 gacaagagca aagtcataaa cggagctctg gaattactca atggtgtcgg tatcgaaggc
     5641 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
     5701 gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat
     5761 acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
     5821 tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
     5881 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
     5941 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
     6001 gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
     6061 tctatgcccc cacttctgag acaagcaatt gagctgttcg accggcaggg agccgaacct
     6121 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
     6181 agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     6241 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     6301 atgctccccg ggtaactaag taaggatccg cggccgcact agaggaattc cgcccctctc
     6361 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgtgtttg
     6421 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     6481 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     6541 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     6601 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     6661 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     6721 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtagtcaac aaggggctga
     6781 aggatgccca gaaggtaccc cattgtatgg gaatctgatc tggggcctcg gtgcacatgc
     6841 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     6901 gttttccttt gaaaaacacg atgataagct taccggtcca ccatgattga acaagatgga
     6961 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     7021 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     7081 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     7141 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     7201 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     7261 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     7321 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     7381 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     7441 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     7501 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     7561 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     7621 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     7681 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgatgt
     7741 acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     7801 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     7861 tgttgttgtt tagtccctcc caattcgata tcaagcttat cgataatcaa cctctggatt
     7921 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     7981 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     8041 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     8101 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     8161 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     8221 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     8281 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     8341 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     8401 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     8461 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgatcgagac
     8521 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     8581 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     8641 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     8701 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
     8761 acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca
     8821 ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc
     8881 aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg
     8941 gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca catggcccga
     9001 gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9061 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9121 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9181 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     9241 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     9301 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     9361 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     9421 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     9481 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     9541 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     9601 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     9661 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     9721 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     9781 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     9841 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     9901 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     9961 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    10021 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10081 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    10141 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    10201 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
    10261 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
    10321 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
    10381 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
    10441 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
    10501 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
    10561 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
    10621 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
    10681 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
    10741 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
    10801 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    10861 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    10921 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    10981 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
    11041 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    11101 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    11161 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
    11221 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    11281 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
    11341 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
    11401 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    11461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    11521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    11581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    11641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    11701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    11761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    11821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    11881 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    11941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12001 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
    12061 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    12121 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
    12181 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
    12241 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
    12301 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
    12361 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    12421 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    12481 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    12541 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    12601 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    12661 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    12721 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    12781 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    12841 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    12901 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    12961 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    13021 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    13081 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    13141 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    13201 gaaaagtgcc acctgac
//
