LOCUS       sequence-12065-e       12370 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector with Tet-based inducible expression of mouse G 
            alpha 12 miR30-based shRNA, constitutive Venus fluorescent protein 
            coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12370)
  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 12370)
  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-G12
FEATURES             Location/Qualifiers
     source          1..12370
                     /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"
     protein_bind    complement(3497..3521)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3568..4779
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4616..4635
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(4904..5521,5522..5527,5528..5563,5564..5566,5567..5602,
                     5603..5605,5606..5641,5642..5650)
                     /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    5688..6262
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5855..5872)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6082..6101
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6290..6292,6293..6295,6296..7009)
                     /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(6335..6356)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6596..6615)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6943..6964
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7058..7646
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7111..7131)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7529..7540)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7649..7665)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7650..7666)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8175..8355
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8381..8398)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8387..8611
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8657..9085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8744..8763)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8954..8975
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9094..9114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9099..9428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9279..9414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9341..9360
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9476..9523
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9542..9916
                     /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    10046..10167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10083..10102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10137..10156
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10216..10232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10216..10232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10229..10251)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10240..10256
                     /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(10264..10270,10271..10288,10289..10294))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10309..10330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10447..10464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10618..11206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10698..10717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11377..12168,12169..12237))
                     /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     12000..12019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12238..12342)
                     /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 gagcgacgac accatcttcg acaacattag tgaagccaca
     3301 gatgtaatgt tgtcgaagat ggtgtcgctg cctactgcct cggaattcaa ggggctactt
     3361 taggagcaat tatcttgttt actaaaactg aataccttgc tatctctttg atacattttt
     3421 acaaagctga attaaaatgg tataaattaa atcacttttt tcaattggaa gactaatgcg
     3481 ttttcgagat atctagaccc agctttcttg tacaaagtgg ttaacatctc gagccataaa
     3541 gatggttaat taacccaccc aagatctggc ctccgcgccg ggttttggcg cctcccgcgg
     3601 gcgcccccct cctcacggcg agcgctgcca cgtcagacga agggcgcagc gagcgtcctg
     3661 atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac
     3721 cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt
     3781 tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg
     3841 gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc
     3901 tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc
     3961 acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg
     4021 tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt
     4081 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat
     4141 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc
     4201 ggcaagaacc caaggtcttg aggccttcgc taatgcggga aagctcttat tcgggtgaga
     4261 tgggctgggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     4321 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     4381 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     4441 gggtggggcc acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     4501 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     4561 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     4621 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     4681 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg
     4741 tccgctaaat tctggccgtt tttggctttt ttgttagacg aagcttggta ccgagctcgg
     4801 atctccaccc cgtaccggtc ctgcaggatt taaatggcgc gccgctagcg ctaccggact
     4861 cagatctcga gctcaagctt cgaattctgc agtcgaattc accatgtcta gactggacaa
     4921 gagcaaagtc ataaacggag ctctggaatt actcaatggt gtcggtatcg aaggcctgac
     4981 gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa
     5041 gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca
     5101 cttctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata
     5161 ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac
     5221 agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc
     5281 cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt
     5341 ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat
     5401 gcccccactt ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt
     5461 ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg
     5521 cgggccgacc gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga
     5581 cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct
     5641 ccccgggtaa ctaagtaagg atccgcggcc gcactagagg aattccgccc ctctccctcc
     5701 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     5761 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     5821 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     5881 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     5941 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     6001 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     6061 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     6121 gcccagaagg taccccattg tatgggattc tgatctgggg cctcggtgca catgctttac
     6181 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     6241 cctttgaaaa acacgatgat aagcttaccg gtacgcgtga tgatccacca tggtgagcaa
     6301 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     6361 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     6421 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     6481 cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     6541 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     6601 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     6661 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     6721 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     6781 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     6841 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     6901 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     6961 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag aattgttgtt
     7021 gtttagtccc tcccaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     7081 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     7141 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     7201 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     7261 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     7321 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     7381 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     7441 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     7501 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     7561 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     7621 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa
     7681 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     7741 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     7801 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     7861 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     7921 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
     7981 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
     8041 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     8101 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
     8161 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     8221 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     8281 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     8341 ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     8401 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     8461 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     8521 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     8581 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     8641 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     8701 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     8761 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     8821 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     8881 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     8941 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     9001 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     9061 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     9121 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     9181 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     9241 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     9301 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     9361 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     9421 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     9481 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     9541 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     9601 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     9661 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     9721 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     9781 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     9841 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     9901 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     9961 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
    10021 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
    10081 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
    10141 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
    10201 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    10261 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    10321 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    10381 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
    10441 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    10501 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    10561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    10621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    10681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    10741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    10801 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    10861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    10921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    10981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    11041 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    11101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    11161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    11221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    11281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
    11341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
    11401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
    11461 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
    11521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
    11581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
    11641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
    11701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
    11761 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    11821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    11881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    11941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    12001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    12061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    12121 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    12181 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    12241 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    12301 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    12361 gccacctgac
//
