LOCUS       sequence-12067-e       12752 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector with Tet-based inducible expression of both mouse 
            G alpha 12 and G alpha 13 miR30-based shRNAs, constitutive Venus 
            fluorescent protein coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12752)
  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 12752)
  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-Venus-TmiR-G13-G12
FEATURES             Location/Qualifiers
     source          1..12752
                     /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"
     ncRNA           3141..3262
                     /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           3290..3304
                     /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           3332..3458
                     /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"
     ncRNA           3523..3644
                     /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           3672..3686
                     /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           3714..3840
                     /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(3879..3903)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3950..5161
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4998..5017
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5286..5903,5904..5909,5910..5945,5946..5948,5949..5984,
                     5985..5987,5988..6023,6024..6032)
                     /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    6070..6644
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6237..6254)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6464..6483
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6672..6674,6675..6677,6678..7391)
                     /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(6717..6738)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6978..6997)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7325..7346
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7440..8028
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7493..7513)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7911..7922)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8031..8047)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8032..8048)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8557..8737
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8763..8780)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8769..8993
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9039..9467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9126..9145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9336..9357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9476..9496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9481..9810
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9661..9796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9723..9742
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9858..9905
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9924..10298
                     /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    10428..10549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10465..10484)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10519..10538
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10598..10614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10598..10614)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10611..10633)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10622..10638
                     /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(10646..10652,10653..10670,10671..10676))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10691..10712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10829..10846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11000..11588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11080..11099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11759..12550,12551..12619))
                     /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     12382..12401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12620..12724)
                     /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 ctagtcgact
     3121 agggataaca gggtaattgt ttgaatgagg cttcagtact ttacagaatc gttgcctgca
     3181 catcttggaa acacttgctg ggattacttc ttcaggttaa cccaacagaa ggctcgagaa
     3241 ggtatattgc tgttgacagt gagcgatggg tgagtctgta aagtatttag tgaagccaca
     3301 gatgtaaata ctttacagac tcacccagtg cctactgcct cggaattcaa ggggctactt
     3361 taggagcaat tatcttgttt actaaaactg aataccttgc tatctctttg atacattttt
     3421 acaaagctga attaaaatgg tataaattaa atcacttttt tcaattggaa gactaatgcg
     3481 ttttcgagat atctagtcga ctagggataa cagggtaatt gtttgaatga ggcttcagta
     3541 ctttacagaa tcgttgcctg cacatcttgg aaacacttgc tgggattact tcttcaggtt
     3601 aacccaacag aaggctcgag aaggtatatt gctgttgaca gtgagcgacg acaccatctt
     3661 cgacaacatt agtgaagcca cagatgtaat gttgtcgaag atggtgtcgc tgcctactgc
     3721 ctcggaattc aaggggctac tttaggagca attatcttgt ttactaaaac tgaatacctt
     3781 gctatctctt tgatacattt ttacaaagct gaattaaaat ggtataaatt aaatcacttt
     3841 tttcaattgg aagactaatg cgttttcgag atatctagac ccagctttct tgtacaaagt
     3901 ggttaacatc tcgagccata aagatggtta attaacccac ccaagatctg gcctccgcgc
     3961 cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac
     4021 gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc
     4081 tcataagact cggccttaga accccagtat cagcagaagg acattttagg acgggacttg
     4141 ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc
     4201 cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag
     4261 gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg
     4321 tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt
     4381 tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta
     4441 gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg
     4501 cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa
     4561 caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg
     4621 gaaagctctt attcgggtga gatgggctgg ggcaccatct ggggaccctg acgtgaagtt
     4681 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     4741 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     4801 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     4861 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     4921 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     4981 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     5041 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     5101 tcagtgttag actagtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     5161 cgaagcttgg taccgagctc ggatctccac cccgtaccgg tcctgcagga tttaaatggc
     5221 gcgccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgaat
     5281 tcaccatgtc tagactggac aagagcaaag tcataaacgg agctctggaa ttactcaatg
     5341 gtgtcggtat cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc
     5401 ctaccctgta ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga
     5461 tgctggacag gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc
     5521 tgcggaacaa cgccaagtca taccgctgtg ctctcctctc acatcgcgac ggggctaaag
     5581 tgcatctcgg cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt
     5641 tcctgtgtca gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc
     5701 actttacact gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag
     5761 agacacctac caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc
     5821 ggcagggagc cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga
     5881 aacagctaaa gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt gacttagaca
     5941 tgctcccagc cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg
     6001 acgattttga ccttgacatg ctccccgggt aactaagtaa ggatccgcgg ccgcactaga
     6061 ggaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
     6121 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     6181 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     6241 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     6301 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     6361 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     6421 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     6481 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat tctgatctgg
     6541 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     6601 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataagcttac cggtacgcgt
     6661 gatgatccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     6721 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     6781 atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag ctgcccgtgc
     6841 cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg
     6901 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     6961 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     7021 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     7081 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     7141 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     7201 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     7261 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
     7321 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     7381 tgtacaagta agaattgttg ttgtttagtc cctcccaatt cgatatcaag cttatcgata
     7441 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     7501 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     7561 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     7621 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     7681 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     7741 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     7801 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     7861 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     7921 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     7981 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     8041 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     8101 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     8161 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     8221 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     8281 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     8341 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     8401 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     8461 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     8521 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     8581 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     8641 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     8701 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
     8761 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     8821 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     8881 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     8941 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     9001 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     9061 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     9121 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     9181 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     9241 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     9301 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     9361 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     9421 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     9481 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     9541 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     9601 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     9661 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     9721 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     9781 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     9841 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     9901 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
     9961 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
    10021 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
    10081 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
    10141 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
    10201 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
    10261 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
    10321 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
    10381 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
    10441 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    10501 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
    10561 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    10621 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    10681 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    10741 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    10801 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    10861 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    10921 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    10981 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    11041 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    11101 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    11161 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    11221 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    11281 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    11341 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    11401 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
    11461 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    11521 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    11581 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
    11641 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
    11701 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
    11761 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
    11821 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
    11881 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
    11941 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
    12001 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    12061 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    12121 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    12181 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    12241 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    12301 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    12361 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    12421 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    12481 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    12541 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    12601 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    12661 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    12721 cgcgcacatt tccccgaaaa gtgccacctg ac
//
