LOCUS       sequence-202030-       13917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation Lentiviral vector for Tet-based inducible shRNA or 
            cDNA expression, constitutive Hygromycin resistance gene 
            coexpression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13917)
  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 13917)
  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-Hygro
FEATURES             Location/Qualifiers
     source          1..13917
                     /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..7061
                     /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             7120..8139
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             7120..8139
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    8605..9193
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8658..8678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9076..9087)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9196..9212)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9197..9213)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9722..9902
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9928..9945)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9934..10158
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10204..10632
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10291..10310)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10501..10522
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10641..10661)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10646..10975
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10826..10961
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10888..10907
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11023..11070
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11089..11463
                     /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    11593..11714
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11630..11649)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11684..11703
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11763..11779)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11763..11779)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11776..11798)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11787..11803
                     /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(11811..11817,11818..11835,11836..11841))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11856..11877
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11994..12011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12165..12753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12245..12264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12924..13715,13716..13784))
                     /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     13547..13566
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13785..13889)
                     /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 gatccgtcga
     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 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     3481 gcgatgagtg gcagggcggg gcgtaaagat ctggatccgg cttactaaaa gccagataac
     3541 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     3601 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
     3661 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     3721 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     3781 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
     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 ctctcccccc 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 tatgggatct gatctggggc
     6961 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     7021 accacgggga cgtggttttc ctttgaaaaa cacgatgata agcttaccgg tacgcgtgat
     7081 agcttgccac aacccgtacc aaagatggat agatccggaa agcctgaact caccgcgacg
     7141 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     7201 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     7261 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     7321 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     7381 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     7441 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     7501 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     7561 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     7621 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     7681 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     7741 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     7801 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     7861 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     7921 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     7981 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     8041 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     8101 aaccgacgcc ccagcactcg tccgagggca aaggaataga gtagatgccg accgaacaag
     8161 agctgatttc gagaacgcct cagccagcaa ctcgcgcgag cctagcaagg caaatgcgag
     8221 agaacggcct tacgcttggt ggcacagttc tcgtccacag ttcgctaagc tcgctcggct
     8281 gggtcgcggg agggccggtc gcagtgattc aggcccttct ggattgtgtt ggtccccagg
     8341 gcacgattgt catgcccacg cactcgggtg atctgactga tcccgcagat tggagatcgc
     8401 cgcccgtgcc tgccgattgg gtgcagatct ctagatctgt acaagtaaag cggccgcgac
     8461 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     8521 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt tagtccctcc
     8581 caattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     8641 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     8701 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     8761 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     8821 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     8881 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     8941 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     9001 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     9061 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     9121 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     9181 gccgcctccc cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat
     9241 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     9301 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     9361 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     9421 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     9481 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     9541 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     9601 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     9661 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     9721 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     9781 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     9841 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     9901 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     9961 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
    10021 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
    10081 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
    10141 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
    10201 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
    10261 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
    10321 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
    10381 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
    10441 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
    10501 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
    10561 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
    10621 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
    10681 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
    10741 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
    10801 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
    10861 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
    10921 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
    10981 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
    11041 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
    11101 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
    11161 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
    11221 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
    11281 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
    11341 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
    11401 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
    11461 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
    11521 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
    11581 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    11641 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    11701 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
    11761 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
    11821 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
    11881 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
    11941 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
    12001 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
    12061 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
    12121 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    12181 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    12241 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    12301 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    12361 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    12421 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    12481 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    12541 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    12601 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    12661 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    12721 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    12781 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    12841 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    12901 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    12961 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    13021 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    13081 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    13141 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    13201 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    13261 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    13321 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    13381 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    13441 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    13501 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    13561 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    13621 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    13681 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    13741 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    13801 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    13861 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
