LOCUS       sequence-352795-       14510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral shRNA vector (pGIPz based) for inducible knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14510)
  AUTHORS   Meerbrey KL, Hu G, Kessler JD, Roarty K, Li MZ, Fang JE, 
            Herschkowitz JI, Burrows AE, Ciccia A, Sun T, Schmitt EM, Bernardi 
            RJ, Fu X, Bland CS, Cooper TA, Schiff R, Rosen JM, Westbrook TF, 
            Elledge SJ
  TITLE     The pINDUCER lentiviral toolkit for inducible RNA interference in 
            vitro and in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3665-70. doi: 
            10.1073/pnas.1019736108. Epub 2011 Feb 9.
  PUBMED    21307310
REFERENCE   2  (bases 1 to 14510)
  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"; doi: 
            "10.1073/pnas.1019736108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2011-03-1- 1"; volume: "108"; issue: "9"; pages: "3665-70"
COMMENT     Sequence Label: pInducer10-mir-RUP-PheS
FEATURES             Location/Qualifiers
     source          1..14510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(15..33,34..56,57..75,76..98,99..117,118..140,141..159,
                     160..182,183..201,202..224,225..243,244..266,267..285)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        355..402
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        394..412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      416..425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             422..796
                     /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"
     misc_feature    861..978
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1276..1320)
                     /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    1505..1738
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2235..2360
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2407..3040
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    3208..3342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3390..3494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3495..3563,3564..4355)
                     /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     complement(3713..3732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4526..5114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5015..5034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5268..5285
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5402..5423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5438..5443,5444..5461,5462..5468)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5476..5492
                     /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)."
     primer_bind     5481..5503
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5500..5516
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5500..5516
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5565..5686
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(6509..6838)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6523..6658
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6577..6596)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6823..6843
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6852..7280)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6962..6983)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7174..7193
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    7326..7550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7539..7556
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             7657..7890
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    8097..8685
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             8203..8214
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     8612..8632
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8707..9309)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8790..8810)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9312..9884
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9471..9490)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     9700..9717
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(join(9933..9941,9942..9977,9978..9980,
                     9981..10016,10017..10019,10020..10055,10056..10061,
                     10062..10679))
                     /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"
     protein_bind    complement(10686..10719)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(10726..11937)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(10870..10889)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     ncRNA           complement(11988..12100)
                     /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"
     CDS             complement(12124..12144)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      13420..13429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(13439..13462)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(13463..13480)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(13501..13517)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     ncRNA           complement(13518..13611)
                     /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"
     CDS             complement(13662..14357)
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     primer_bind     complement(14433..14457)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(14454..14492)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 tcgactttca cttttctcta tcactgatag ggagtggtaa actcgacttt cacttttctc
       61 tatcactgat agggagtggt aaactcgact ttcacttttc tctatcactg atagggagtg
      121 gtaaactcga ctttcacttt tctctatcac tgatagggag tggtaaactc gactttcact
      181 tttctctatc actgataggg agtggtaaac tcgactttca cttttctcta tcactgatag
      241 ggagtggtaa actcgacttt cacttttctc tatcactgat agggagtggt aaaggatcca
      301 gtcgactgaa ttggttcctt taagctagcc tagaacctcg gacctgcagc acgtgttgac
      361 aattaatcat cggcatagta tatcggcata gtataatacg actcactata ggagggccac
      421 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
      481 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
      541 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
      601 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
      661 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
      721 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
      781 cgaggagcag gactgattcc ggactagagc gggcccggta ctaaccaaac tggatctctg
      841 ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg taattcttta
      901 gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta ccccccaatc
      961 cccccttttc ttttaaaatt gtggatgaat actgccattt gtctgcagaa ttggcgcacg
     1021 cagtgccgat ccgttcacta atcgaatgga tctgtctctg tctctctctc caccttcttc
     1081 ttctattcct tcgggcctgt cgggtcccct cggggttggg aggtgggtct gaaacgataa
     1141 tggtgaatat ccctgcctaa ctctattcac tatagaaagt acagcaaaaa ctattcttaa
     1201 acctaccaag cctcctacta tcattatgaa taattttata taccacagcc aatttgttat
     1261 gttaaaccaa ttccacaaac ttgcccattt atctaattcc aataattctt gttcattctt
     1321 ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt aagcttgtgt aattgttaat
     1381 ttctctgtcc cactccatcc aggtcgtgtg attccaaatc tgttccagag atttattact
     1441 ccaactagca ttccaaggca cagcagtggt gcaaatgagt tttccagagc aaccccaaat
     1501 ccccaggagc tgttgatcct ttaggtatct ttccacagcc aggattcttg cctggagctg
     1561 cttgatgccc cagactgtga gttgcaacag atgctgttgc gcctcaatag ccctcagcaa
     1621 attgttctgc tgctgcacta taccagacaa taattgtctg gcctgtaccg tcagcgtcat
     1681 tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac ccaaggaaca aagctcctat
     1741 tcccactgct cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc
     1801 taatggttca atttttacta ctttatattt atataattca cttctccaat tgtccctcat
     1861 atctcctcct ccaggtctga agatcagcgg ccggccgctt gctgtgcggt ggtcttactt
     1921 ttgttttgct cttcctctat cttgtctaaa gcttccttgg tgtcttttat ctctatcctt
     1981 tgatgcacac aatagagggt tgctactgta ttatataatg atctaagttc ttctgatcct
     2041 gtctgaaggg atggttgtag ctgtcccagt atttgtctac agccttctga tgtttctaac
     2101 aggccaggat taactgcgaa tcgttctagc tccctgcttg cccatactat atgttttaat
     2161 ttatattttt tctttccccc tggccttaac cgaatttttt cccatcgcga tctaattctc
     2221 ccccgcttaa tactgacgct ctcgcaccca tctctctcct tctagcctcc gctagtcaaa
     2281 atttttggcg tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc
     2341 cgagtcctgc gtcgagagag ctcctctggt ttccctttcg ctttcaagtc cctgttcggg
     2401 cgccactgct agagattttc cacactgact aaaagggtct gagggatctc tagttaccag
     2461 agtcacacaa cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct
     2521 taagcagtgg gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga
     2581 gagacccagt acaggcaaaa agcagctgct tatatgcagg atctgagggc tcgccactcc
     2641 ccagtcccgc ccaggccacg cctccctgga aagtccccag cggaaagtcc cttgtagcaa
     2701 gctcgatatc agcagttctt gaagtactcc ggatgcagct ctcgggccac gtgatgaaat
     2761 gctaggcggc tgtcaaacct ccactctaac acttctctct ccgggtcatc catcccatgc
     2821 aggctcacag ggtgtaacaa gctggtgttc tctcctttat tggcctcttc taccttatct
     2881 ggctcaactg gtactagctt gtagcaccat ccaaaggtca gtggatatct gacccctggc
     2941 cctggtgtgt agttctgcta atcagggaag tagccttgtg tgtggtagat ccacagatca
     3001 aggatatctt gtcttctttg ggagtgaatt agcccttcca actactaagt ttgtagtaca
     3061 tatttaacaa atacaatttc tttaaaatga aaataattca gaggaatcac aggtttagag
     3121 taaatgaaac cacaggtaat tggcagtggt aatagggtat ggggtgggaa gtttgggatg
     3181 attttggtta gcttgagtta tccagttgat ccagacatga taagatacat tgatgagttt
     3241 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     3301 attgctttat ttgtaaccat tataagctgc aataaacaag ttgatctccc gatccgtcga
     3361 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3481 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3541 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3601 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3661 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3721 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3781 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3841 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3901 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3961 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4021 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4081 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4141 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4201 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4261 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4321 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4381 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4441 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttagtgca
     4861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5161 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5221 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5281 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     5341 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     5401 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     5461 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     5521 attacgccaa gctctagcta gaggtcgacg gtatacagac atgataagat acattgatta
     5581 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     5641 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga
     5701 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
     5761 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtagc acgtgctatt
     5821 cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac
     5881 agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg
     5941 ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc
     6001 cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc
     6061 gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac
     6121 aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca
     6181 tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg
     6241 agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca
     6301 gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt
     6361 gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga
     6421 ttccttgcgg tcacgtcgcg gtgagttcag gctttttcat cacgtgctga tcagatccga
     6481 aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca aaaaagcctc
     6541 ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca taaataaaaa
     6601 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     6661 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     6721 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     6781 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     6841 tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     6901 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     6961 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     7021 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     7081 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     7141 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     7201 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     7261 tgcgccgcta cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc
     7321 agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc
     7381 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     7441 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     7501 aggcacgggg aaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga
     7561 tcagcgggtt taaacaatgt caaggcctct cactctctga tattcatttc tttgcaagtt
     7621 ataaatactg aataataaga tgacatgaac tactactgct agagattttc cacactgact
     7681 aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt
     7741 gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga
     7801 gctcccaggc tcagatctgg tctaaccaga gagacccagt acaagcaaaa agcagatctt
     7861 gtcttcgttg ggagtgaatt agcccttcca gtcccctctt ttcttttaaa aagtggctaa
     7921 gatctacagc tgccttgtaa gtcattggtc ttaaaggtac ctgaggtgtg actggaaaac
     7981 ccacctcctc ctcctcttgt gcttctagcc aggcacaatc agcattggta gctgctgtat
     8041 tgctacttgt gattgctcca tgtttttcta ggtctcgact gcagaattaa ttccaggcgg
     8101 ggaggcggcc caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg
     8161 ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac
     8221 gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccatggaaa
     8281 ggacgtcagc ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt
     8341 ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc
     8401 gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc
     8461 agcaaacaca gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag
     8521 agacagcaac caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc
     8581 atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag
     8641 aataccagtc aatctttcac aaattttgta atccagaggt tgattgttcc agacgcgctt
     8701 aattaatcag gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc
     8761 gacgtcggcg gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga
     8821 ggtctccagg aaggcgggca ccccagcgcg ctcggccgcc tccactccgg ggagcacgac
     8881 ggcgctgccc agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca
     8941 cgcgggctcc ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag
     9001 ccgggaaccg ctcaactcgg ccatgcgcga gccgatctcg gcgaacaccg cccccgcttc
     9061 gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt
     9121 gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat
     9181 gtggcggtcc gggtcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
     9241 ggtgcgtacg gcccggggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
     9301 ggtggccatg gtattatcgt gtttttcaaa ggaaaaccac gtccccgtgg ttcggggggc
     9361 ctagacgttt ttttaacctc gactaaacac atgtaaagca tgtgcaccga ggccccagat
     9421 cagatcccat acaatggggt accttctggg catccttcag ccccttgttg aatacgcttg
     9481 aggagagcca tttgactctt tccacaacta tccaactcac aacgtggcac tggggttgtg
     9541 ccgcctttgc aggtgtatct tatacacgtg gcttttggcc gcagaggcac ctgtcgccag
     9601 gtggggggtt ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt caagaagctt
     9661 ccagaggaac tgcttccttc acgacattca acagaccttg cattcctttg gcgagagggg
     9721 aaagacccct aggaatgctc gtcaagaaga cagggccagg tttccgggcc ctcacattgc
     9781 caaaagacgg caatatggtg gaaaataaca tatagacaaa cgcacaccgg ccttattcca
     9841 agcggcttcg gccagtaacg ttaggggggg gggagggaga ggggcggaat tggcgcgcca
     9901 tacttcgtat aatgtatgct atacgaagtt atttacccgg ggagcatgtc aaggtcaaaa
     9961 tcgtcaagag cgtcagcagg cagcatatca aggtcaaagt cgtcaagggc atcggctggg
    10021 agcatgtcta agtcaaaatc gtcaagggcg tcggtcggcc cgccgctttc gcactttagc
    10081 tgtttctcca ggccacatat gattagttcc aggccgaaaa ggaaggcagg ttcggctccc
    10141 tgccggtcga acagctcaat tgcttgtctc agaagtgggg gcatagaatc ggtggtaggt
    10201 gtctctcttt cctcttttgc tacttgatgc tcctgttcct ccaatacgca gcccagtgta
    10261 aagtggccca cggcggacag agcgtacagt gcgttctcca gggagaagcc ttgctgacac
    10321 aggaacgcga gctgattttc cagggtttcg tactgtttct ctgttgggcg ggtgccgaga
    10381 tgcactttag ccccgtcgcg atgtgagagg agagcacagc ggtatgactt ggcgttgttc
    10441 cgcagaaagt cttgccatga ctcgccttcc agggggcaga agtgggtatg atgcctgtcc
    10501 agcatctcga ttggcagggc atcgagcagg gcccgcttgt tcttcacgtg ccagtacagg
    10561 gtaggctgct caactcccag cttttgagcg agtttccttg tcgtcaggcc ttcgataccg
    10621 acaccattga gtaattccag agctccgttt atgactttgc tcttgtccag tctagacatg
    10681 gtggcataac ttcgtatagc atacattata cgaagttatg gatccgtcta acaaaaaagc
    10741 caaaaacggc cagaatttag cggacaattt actagtctaa cactgaaaat tacatattga
    10801 cccaaatgat tacatttcaa aaggtgccta aaaaacttca caaaacacac tcgccaaccc
    10861 cgagcgcata gttcaaaacc ggagcttcag ctacttaaga agataggtac ataaaaccga
    10921 ccaaagaaac tgacgcctca cttatccctc ccctcaccag aggtccggcg cctgtcgatt
    10981 caggagagcc taccctaggc ccgaaccctg cgtcctgcga cggagaaaag cctaccacac
    11041 acctaccggc aggtggcccc accctgcatt ataagccaac agaacgggtg acgtcacgac
    11101 acgacgaggg cgcgcgctcc caaaggtacg ggtgcactgc ccaacggcac cgccataact
    11161 gccgcccccg caacagacga caaaccgagt tctccagtca gtgacaaact tcacgtcagg
    11221 gtccccagat ggtgccccag cccatctcac ccgaataaga gctttcccgc attagcgaag
    11281 gcctcaagac cttgggttct tgccgcccac catgcccccc accttgtttc aacgacctca
    11341 cagcccgcct cacaagcgtc ttccattcaa gactcgggaa cagccgccat tttgctgcgc
    11401 tccccccaac ccccagttca gggcaacctt gctcgcggac ccagactaca gcccttggcg
    11461 gtctctccac acgcttccgt cccaccgagc ggcccggcgg ccacgaaagc cccggccagc
    11521 ccagcagccc gctactcacc aagtgacgat cacagcgatc cacaaacaag aaccgcgacc
    11581 caaatcccgg ctgcgacgga actagctgtg ccacacccgg cgcgtcctta tataatcatc
    11641 ggcgttcacc gccccacgga gatccctccg cagaatcgcc gagaagggac tacttttcct
    11701 cgcctgttcc gctctctgga aagaaaacca gtgccctaga gtcacccaag tcccgtccta
    11761 aaatgtcctt ctgctgatac tggggttcta aggccgagtc ttatgagcag cgggccgctg
    11821 tcctgagcgt ccgggcggaa ggatcaggac gctcgctgcg cccttcgtct gacgtggcag
    11881 cgctcgccgt gaggaggggg gcgcccgcgg gaggcgccaa aacccggcgc ggaggccaga
    11941 tcttacgcgt cgccgcgtgt ttaaacgcat tagtcttcca attgaaaaaa gtgatttaat
    12001 ttataccatt ttaattcagc tttgtaaaaa tgtatcaaag agatagcaag gtattcagtt
    12061 ttagtaaaca agataattgc tcctaaagta gccccttgaa ttcgtaatag atccacggag
    12121 atcaccctga aaatacaaat tctcagaatt gtgagatcta gggataacag ggtaatccgc
    12181 ggtcgacggt atcggggacc aaaatggcaa gtaaaatagc ctgatgggat aggctctaag
    12241 tccaacgaac cagtgtcacc actgacacaa tgaggaaaac catgtcacat ctcgcagaac
    12301 tggttgccag tgcgaaggcg gccattagcc aggcgtcaga tgttgccgcg ttagataatg
    12361 tgcgcgtcga atatttgggt aaaaaagggc acttaaccct tcagatgacg accctgcgtg
    12421 agctgccgcc agaagagcgt ccggcagctg gtgcggttat caacgaagcg aaagagcagg
    12481 ttcagcaggc gctgaatgcg cgtaaagcgg aactggaaag cgctgcactg aatgcgcgtc
    12541 tggcggcgga aacgattgat gtctctctgc caggtcgtcg cattgaaaac ggcggtctgc
    12601 atccggttac ccgtaccatc gaccgtatcg aaagtttctt cggtgagctt ggctttaccg
    12661 tggcaaccgg gccggaaatc gaagacgatt atcataactt cgatgctctg aacattcctg
    12721 gtcaccaccc ggcgcgcgct gaccacgaca ctttctggtt tgacactacc cgcctgctgc
    12781 gtacccagac ctctggcgta cagatccgca ccatgaaagc ccagcagcca ccgattcgta
    12841 tcatcgcgcc tggccgtgtt tatcgtaacg actacgacca gactcacacg ccgatgttcc
    12901 atcagatgga aggtctgatt gttgatacca acatcagctt taccaacctg aaaggcacgc
    12961 tgcacgactt cctgcgtaac ttctttgagg aagatttgca gattcgcttc cgtccttcct
    13021 acttcccgtt taccgaacct tctgcagaag tggacgtcat gggtaaaaac ggtaaatggc
    13081 tggaagtgct gggctgcggg atggtgcatc cgaacgtgtt gcgtaacgtt ggcatcgacc
    13141 cggaagttta ctctggtttc ggcttcggga tggggatgga gcgtctgact atgttgcgtt
    13201 acggcgtcac cgacctgcgt tcattcttcg aaaacgatct gcgtttcctc aaacagttta
    13261 aataaggcag gaatagatta tgaaattcag tgaactgtgg ttacgcgaat gggtgaaccc
    13321 ggcgattgat agcgatgcgc tggcaaatca aatcactatg gcgggcctgg aagttggggg
    13381 atcgatccgg taccattacc ctgttatccc tatctagacc catggtggcg tctccttcgg
    13441 gcccctggaa gagaacttcc agacttccac ggggtaccag catatggacg tcaagcttat
    13501 cgataccgtc gacctcgagc cttctgttgg gttaacctga agaagtaatc ccagcaagtg
    13561 tttccaagat gtgcaggcaa cgattctgta aagtactgaa gcctcattca acatagtata
    13621 tgtgctgccg aagcgagcac ttaacaaggc ttgcggccgc atcatctgtg ccccagtttg
    13681 ctagggaggt cgcagtactt ggccacagcc atctcgtgct gctcgacgta ggtctctttg
    13741 tcggcctcct tgattctttc cagtctgtgg tccacgaagt ggaagccggg catcttgagg
    13801 ttcttagcgg gtttcttgga tctgtatgtg gtcttgaagg agcagtgcag gtagcccccg
    13861 cccacgagct tcagggccat ctggctgtgg cctctcaggc cgccgtcagc ggggtacagc
    13921 atctcggtgt tggcctccca gccgcgtgtt ttcttctgca tcacagggcc gttggatggg
    13981 aagttcaccc cgttgatctt gacgttgtag atgatgcagc cgttctggaa gctggtgtcc
    14041 tgggtagcgg tcagcacgcc cccgtcttcg tatgtggtga ttctctccca tgtgaagccc
    14101 tcagggaagg actgcttaaa gaagtcgggg atgccctggg tgtggttgat gaaggctttg
    14161 ctgccgtaca tgaagctggt agccaggatg tcgaaggcga aggggagagg gccgccctcg
    14221 accaccttga tcttcatggt ctgggtgccc tcgtagggct tgccttcgcc ctcggatgtg
    14281 cacttgaagt ggtggttgtt cacggtgccc tccatgtaca gcttcatgtg catgttctcc
    14341 ttgatcagct cgctcatggt ggcgaccggt aggctggatc ggtcccggtg tcttctatgg
    14401 aggtcaaaac agcgtggatg gcgtctccag gcgatctgac ggttcactaa acgagctctg
    14461 cttatatagg cctcccaccg tacacgccta cctcgacccg ggtaccgagc
//
