LOCUS       sequence-438025-       10273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To inducibly knockdown NEK4 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10273)
  TITLE     Lee Lab shRNA knockdown vectors
REFERENCE   2  (bases 1 to 10273)
  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"
COMMENT     Sequence Label: LT3GEPIR-NEK4shRNA2
FEATURES             Location/Qualifiers
     source          1..10273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    147..168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(183..188,189..206,207..213)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    221..237
                     /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     226..248
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     245..261
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     245..261
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     280..300
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        282..300
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        328..554
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             555..735
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    782..907
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1400..1633
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1818..1862
                     /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    2129..2246
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2262..2640
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2270..2288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2306..2324
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2342..2360
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2378..2396
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2414..2432
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2450..2468
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2486..2504
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2549..2573
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2664..2666,2667..2669,2670..3383)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2709..2730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2970..2989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3317..3338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           3577..3591
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        3771..4270
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3806..3829)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4203..4222
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4291..4890
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4291..4310)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4787..4807
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4951..5518
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4968..5520
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5112..5129)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5339..5358
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5532..6149,6150..6155,6156..6191,6192..6194,6195..6230,
                     6231..6233,6234..6269,6270..6278)
                     /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    6324..6912
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6377..6397)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6795..6806)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7119..7352
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7424..7545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7461..7480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7515..7534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7585..7720
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7740..7759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7741..7759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7769..7786)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7769..7785)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7778..7800)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7927..8382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8014..8033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8224..8245
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8408..8512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8513..8581,8582..9373)
                     /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(8731..8750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9544..10132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10033..10052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
       61 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      121 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      181 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      241 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
      301 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
      361 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
      421 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
      481 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
      541 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
      601 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
      661 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
      721 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
      781 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
      841 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
      901 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
      961 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     1021 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     1081 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     1141 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     1201 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     1261 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     1321 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     1381 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     1441 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     1501 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     1561 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     1621 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     1681 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     1741 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     1801 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     1861 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     1921 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     1981 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     2041 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     2101 tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt ggggggtaca
     2161 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     2221 aacaaattac aaaaattcaa aattttatcg attctagatt cgagtttact ccctatcagt
     2281 gatagagaac gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta
     2341 ctccctatca gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt
     2401 atgaccagtt tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt
     2461 gatagagaac gtatatccag tttactccct atcagtgata gagaacgtat aagctttagg
     2521 cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt gaaccgtcag atcgcctgga
     2581 gcaattccac aacacttttg tcttatacca actttccgta ccacttccta ccctcgtaaa
     2641 gtcgagcttg cgttggatcc accatggtga gcaagggcga ggagctgttc accggggtgg
     2701 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtctggcg
     2761 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2821 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2881 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2941 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     3001 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     3061 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     3121 tcatggccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg
     3181 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     3241 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     3301 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3361 gcatggacga gctgtacaag taatgacgta cggcggccgc ctcgactagg gataacaggg
     3421 taattgtttg aatgaggctt cagtacttta cagaatcgtt gcctgcacat cttggaaaca
     3481 cttgctggga ttacttcgac ttcttaaccc aacagaaggc tcgagaaggt atattgctgt
     3541 tgacagtgag cgagattact gtttactgag cctttagtga agccacagat gtaaaggctc
     3601 agtaaacagt aatcgtgcct actgcctcgg acttcaaggg gtctagaatt cgagcaatta
     3661 tcttgtttac taaaactgaa taccttgcta tctctttgat acatttttac aaagctgaat
     3721 taaaatggta taaattaaat cacttttttc aattgacgcg taattctacc gggtagggga
     3781 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     3841 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     3901 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     3961 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     4021 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     4081 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     4141 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     4201 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     4261 gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc
     4321 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4381 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4441 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4501 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4561 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     4621 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4681 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     4741 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4801 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     4861 tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc
     4921 gaaaggagcg cacgacccca tgcatccaat tccgcccccc ccccctaacg ttactggccg
     4981 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     5041 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     5101 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     5161 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     5221 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     5281 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     5341 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     5401 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     5461 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     5521 tggccacaac catgagtaga ctggacaaga gcaaagtcat aaacggagct ctggaattac
     5581 tcaatggtgt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg
     5641 agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa
     5701 tcgagatgct ggacaggcat catacccact tctgccccct ggaaggcgag tcatggcaag
     5761 actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg
     5821 ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc
     5881 tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg
     5941 tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg
     6001 aaagagagac acctaccacc gattctatgc ccccacttct gagacaagca attgagctgt
     6061 tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc
     6121 tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact
     6181 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     6241 ctcttgacga ttttgacctt gacatgctcc ccgggtaatc cggactcgac ctgcagccaa
     6301 gcttatcgaa acgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga
     6361 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     6421 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     6481 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     6541 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     6601 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6661 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     6721 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6781 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6841 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     6901 ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac
     6961 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     7021 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     7081 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     7141 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     7201 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     7261 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     7321 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     7381 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     7441 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     7501 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     7561 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     7621 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     7681 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     7741 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     7801 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     7861 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     7921 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     7981 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     8041 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     8101 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     8161 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     8221 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     8281 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     8341 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     8401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     8461 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     8521 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     8581 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     8641 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     8701 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     8761 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8821 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8881 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8941 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     9001 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     9061 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     9121 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     9181 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     9241 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     9301 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     9361 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     9421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     9481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     9541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     9601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     9661 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     9721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     9841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     9901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     9961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
    10021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
    10081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
    10141 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
    10201 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
    10261 tcgccgcagc cga
//
