LOCUS       sequence-12059-e       13170 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral vector for Tet-based inducible shRNA or cDNA expression, 
            constitutive Venus fluorescent protein coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13170)
  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 13170)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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
FEATURES             Location/Qualifiers
     source          1..13170
                     /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"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2614..2738
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2763..2768,2769..2786,2787..2793)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2847..3506
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2913..2932)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3848..4153
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4032..4051
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4194..4318)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4368..5579
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5416..5435
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5704..6321,6322..6327,6328..6363,6364..6366,6367..6402,
                     6403..6405,6406..6441,6442..6450)
                     /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    6488..7062
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6655..6672)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6882..6901
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(7090..7092,7093..7095,7096..7809)
                     /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(7135..7156)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7396..7415)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7743..7764
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7858..8446
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7911..7931)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8329..8340)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8449..8465)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8450..8466)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8975..9155
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9181..9198)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9187..9411
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9457..9885
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9544..9563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9754..9775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9894..9914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9899..10228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10079..10214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10141..10160
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10276..10323
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10342..10716
                     /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    10846..10967
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10883..10902)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10937..10956
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11016..11032)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11016..11032)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11029..11051)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11040..11056
                     /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(11064..11070,11071..11088,11089..11094))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11109..11130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11247..11264)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11418..12006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11498..11517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12177..12968,12969..13037))
                     /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     12800..12819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13038..13142)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaattc tagacagtta tcaacaagtt tgtacaaaaa agctgaacga
     2641 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     2701 acataatact gtaaaacaca acatatccag tcactatggc ggccgcatta ggcaccccag
     2761 gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag gatccggcga
     2821 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     2881 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     2941 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     3001 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     3061 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     3121 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     3181 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     3241 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     3301 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     3361 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     3421 atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     3481 gcgatgagtg gcagggcggg gcgtaaagat ctggatccgg cttactaaaa gccagataac
     3541 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     3601 gaagtatgtc aaaaagaggt gtgctatgaa gcagcgtatt acagtgacag ttgacagcga
     3661 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     3721 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     3781 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacagggac
     3841 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     3901 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
     3961 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     4021 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     4081 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     4141 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc gaccatagtg
     4201 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     4261 atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac aaagtggttg
     4321 ataacatctc gagccataaa gatggttaat taacccaccc aagatctggc ctccgcgccg
     4381 ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca cgtcagacga
     4441 agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
     4501 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
     4561 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
     4621 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
     4681 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
     4741 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
     4801 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
     4861 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
     4921 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
     4981 aggtgggggg catggtgggc ggcaagaacc caaggtcttg aggccttcgc taatgcggga
     5041 aagctcttat tcgggtgaga tgggctgggg caccatctgg ggaccctgac gtgaagtttg
     5101 tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg
     5161 ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg
     5221 ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc
     5281 tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg
     5341 acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct
     5401 atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt
     5461 tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc
     5521 agtgttagac tagtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagacg
     5581 aagcttggta ccgagctcgg atctccaccc cgtaccggtc ctgcaggatt taaatggcgc
     5641 gccgctagcg ctaccggact cagatctcga gctcaagctt cgaattctgc agtcgaattc
     5701 accatgtcta gactggacaa gagcaaagtc ataaacggag ctctggaatt actcaatggt
     5761 gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct
     5821 accctgtact ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc aatcgagatg
     5881 ctggacaggc atcataccca cttctgcccc ctggaaggcg agtcatggca agactttctg
     5941 cggaacaacg ccaagtcata ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg
     6001 catctcggca cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc
     6061 ctgtgtcagc aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac
     6121 tttacactgg gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga ggaaagagag
     6181 acacctacca ccgattctat gcccccactt ctgagacaag caattgagct gttcgaccgg
     6241 cagggagccg aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa
     6301 cagctaaagt gcgaaagcgg cgggccgacc gacgcccttg acgattttga cttagacatg
     6361 ctcccagccg atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac
     6421 gattttgacc ttgacatgct ccccgggtaa ctaagtaagg atccgcggcc gcactagagg
     6481 aattccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     6541 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     6601 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     6661 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     6721 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     6781 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     6841 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     6901 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggattc tgatctgggg
     6961 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     7021 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aagcttaccg gtacgcgtga
     7081 tgatccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     7141 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     7201 gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc
     7261 tggcccaccc tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac
     7321 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     7381 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     7441 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     7501 ctggggcaca agctggagta caactacaac agccacaacg tctatatcac cgccgacaag
     7561 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     7621 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     7681 gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     7741 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     7801 tacaagtaag aattgttgtt gtttagtccc tcccaattcg atatcaagct tatcgataat
     7861 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     7921 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     7981 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     8041 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     8101 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     8161 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     8221 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     8281 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     8341 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     8401 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
     8461 cctcgatcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     8521 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     8581 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     8641 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     8701 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg
     8761 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa
     8821 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg
     8881 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca
     8941 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg
     9001 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     9061 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     9121 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
     9181 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     9241 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     9301 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     9361 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     9421 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     9481 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     9541 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     9601 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     9661 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     9721 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     9781 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     9841 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     9901 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     9961 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
    10021 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
    10081 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
    10141 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
    10201 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
    10261 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
    10321 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
    10381 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
    10441 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
    10501 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
    10561 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
    10621 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
    10681 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
    10741 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
    10801 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
    10861 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    10921 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
    10981 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
    11041 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
    11101 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
    11161 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
    11221 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    11281 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
    11341 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
    11401 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    11461 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    11521 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    11581 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    11641 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    11701 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    11761 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    11821 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    11881 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    11941 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    12001 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    12061 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    12121 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    12181 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    12241 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    12301 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    12361 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    12421 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    12481 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    12541 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    12601 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    12661 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    12721 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    12781 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    12841 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    12901 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    12961 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    13021 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    13081 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    13141 cgcacatttc cccgaaaagt gccacctgac
//
