LOCUS       sequence-491498-       10307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NFKB-dependent expression of a bicistronic cDNA encoding a suicide 
            gene (modified deoxycytidine kinase) and EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10307)
  AUTHORS   Lampson BL, Ramiotarez AS, Baro M, He L, Hegde M, Koduri V, Pfaff 
            JL, Hanna RE, Kowal J, Shirole NH, He Y, Doench JG, Contessa JN, 
            Locher KP, Kaelin WG Jr
  TITLE     Positive selection CRISPR screens reveal a druggable pocket in an 
            oligosaccharyltransferase required for inflammatory signaling to 
            NF-kappaB.
  JOURNAL   Cell. 2024 Apr 25;187(9):2209-2223.e16. doi: 
            10.1016/j.cell.2024.03.022.
  PUBMED    38670073
REFERENCE   2  (bases 1 to 10307)
  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: 
            "10.1016/j.cell.2024.03"; journalName: "Cell"; date: "2024-04-25- 
            25"; volume: "187"; issue: "9"; pages: "2209-2223.e16"
COMMENT     Sequence Label: NFKB3-CK-IRES-GFP hygro
FEATURES             Location/Qualifiers
     source          1..10307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(245..744)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(293..312)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     686..709
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     CDS             complement(join(806..1519,1520..1522,1523..1525))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(809..1519,1520..1522,1523..1525))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(851..872)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1200..1219
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1459..1480
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    join(1526..1534,1535..1537,1538..2112)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1699..1718)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     1928..1945
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2155..2196)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3059..3070
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    3319..3436
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(3703..3747)
                     /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    3932..4165
                     /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    4658..4783
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4830..5010
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(5011..5237)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(5265..5285)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5265..5283)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5304..5320)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5304..5320)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5317..5339)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5328..5344
                     /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(5352..5358,5359..5376,5377..5382))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5397..5418
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5535..5552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5706..6294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5786..5805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6465..7256,7257..7325))
                     /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     7088..7107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7326..7430)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7456..7911)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7593..7614)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7805..7824
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8038..8060
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8052..8069
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8053..8069
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8079..8098
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8079..8097
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      8118..8253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8293..8414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8304..8323)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8358..8377
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             8486..8719
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    8801..9389
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             8907..8918
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     9316..9336
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    9445..9478
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(9509..10307,1..227))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(join(9509..10307,1..227))
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
ORIGIN
        1 tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc tttgtagaaa
       61 ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa
      121 gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg
      181 atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatggt ggcgggatgc
      241 aggtcgaaag gcccggagat gaggaagagg agaacagcgc ggcagacgtg cgcttttgaa
      301 gcgtgcagaa tgccgggcct ccggaggacc ttcgggcgcc cgccccgccc ctgagcccgc
      361 ccctgagccc gcccccggac ccaccccttc ccagcctctg agcccagaaa gcgaaggagc
      421 aaagctgcta ttggccgctg ccccaaaggc ctacccgctt ccattgctca gcggtgctgt
      481 ccatctgcac gagactagtg agacgtgcta cttccatttg tcacgtcctg cacgacgcga
      541 gctgcggggc gggggggaac ttcctgacta ggggaggagt agaaggtggc gcgaaggggc
      601 caccaaagaa cggagccggt tggcgcctac cggtggatgt ggaatgtgtg cgaggccaga
      661 ggccacttgt gtagcgccaa gtgcccagcg gggctgctaa agcgcatgct ccagactgcc
      721 ttgggaaaag cgcctcccct acccggtaga attcgttaac ctcgagggcc agccttaatt
      781 aagtttaaac gcggaattcg tcgacttact tgtacagctc gtccatgccg agagtgatcc
      841 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
      901 tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga
      961 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc
     1021 ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt
     1081 ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga
     1141 tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg
     1201 tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca
     1261 tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca
     1321 cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc
     1381 agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc
     1441 tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga
     1501 acagctcctc gcccttgctc accatggttg tggccatatt atcatcgtgt ttttcaaagg
     1561 aaaaccacgt ccccgtggtt cggggggcct agacgttttt ttaacctcga ctaaacacat
     1621 gtaaagcatg tgcaccgagg ccccagatca gatcccatac aatggggtac cttctgggca
     1681 tccttcagcc ccttgttgaa tacgcttgag gagagccatt tgactctttc cacaactatc
     1741 caactcacaa cgtggcactg gggttgtgcc gcctttgcag gtgtatctta tacacgtggc
     1801 ttttggccgc agaggcacct gtcgccaggt ggggggttcc gctgcctgca aagggtcgct
     1861 acagacgttg tttgtcttca agaagcttcc agaggaactg cttccttcac gacattcaac
     1921 agaccttgca ttcctttggc gagaggggaa agacccctag gaatgctcgt caagaagaca
     1981 gggccaggtt tccgggccct cacattgcca aaagacggca atatggtgga aaataacata
     2041 tagacaaacg cacaccggcc ttattccaag cggcttcggc cagtaacgtt aggggggggg
     2101 gagggagagg gggggggggg gggcggaatt tacgtagcgg ctagctcatt actacgtaga
     2161 atcgagaccg aggagagggt tagggatagg cttaccgctc gcgccagacg ccccacttgc
     2221 tcccaaagta ctcaaaaact ctttgacctt ttcaaccaga ctttcatatt tgtctttaaa
     2281 gtcttcatta acatccagtg ttaagatagg cacctcttga agataatcga agttggtttt
     2341 cagtgtccta tgcaggagcc agctttcatg tttataatga agcttctcta aatattcaag
     2401 aggaatgcct tgctcttcat ttcttccccg taaatatatt ctatgtaagc atgtctctgg
     2461 agtggcttga agataaatga ttccatccaa ttcaaggctt tggccaaatt ggttattcat
     2521 ccagtcatgc cagtcttgat aaattgtcca ctctgtctca ttcatgcatt cagattcata
     2581 caaattagat gcaaaaatat acctcgcact atacacagat cgttcaaaaa ataatacagg
     2641 tttctctgca tctttgagct tgccattcag agaggcaagc tgagctctta tcatactgag
     2701 acaggcgtag gtttggaagg taaaagacca tcgttcaggt ttctcataca tcatctgaag
     2761 aacattccca ccatttttct gctccattgt aagttcctca aattcatctt gagtactttg
     2821 aacattgcac catctggcaa caggttcagg aaccacttcc caatcttcac acaattgttt
     2881 aaggatattc acaaatgttg acttccctgc agcgatgttc ccttcgatgg agattttctt
     2941 gatgcgggtc ccctcagagc tggctgagaa agatgggcag cttctcttgg gcggggtggc
     3001 cattttacca acagtaccgg aatgccaagc ttacttagat cctctagaga gacactggtg
     3061 ggagctgcag agcagagctg ggtgggagcg cggtcactgc tcgttcgcgg aaccgccggc
     3121 tctatccagt cttctcagtt gctcgctgca gtcgcggttg gagtagtagg cgcctcagct
     3181 ggcgccttta tagaagcgct gtgaatggat ggacggtcga gatggggaat ccccagcctc
     3241 gagatgggga atccccagcc tcgagatggg gaatccccag cctcgaggat gggaatccga
     3301 attccaccac actggaataa aattttgaat ttttgtaatt tgtttttgta attctttagt
     3361 ttgtatgtct gttgctatta tgtctactat tctttcccct gcactgtacc ccccaatccc
     3421 cccttttctt ttaaaagtta accgataccg tcgagatccg ttcactaatc gaatggatct
     3481 gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg gtcccctcgg
     3541 ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat
     3601 agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa
     3661 ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc
     3721 taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga
     3781 gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt
     3841 ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag cagtggtgca
     3901 aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc
     3961 cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg
     4021 ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa
     4081 ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc
     4141 caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct
     4201 tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata
     4261 taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga tcagcggccg
     4321 cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct
     4381 tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata
     4441 atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc
     4501 tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct agctccctgc
     4561 ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt
     4621 tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct
     4681 ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc
     4741 ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt
     4801 tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg
     4861 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac
     4921 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     4981 ggctcagatc tggtctaacc agagagaccc gtttatgtat cgagctaggc acttaaatac
     5041 aatatctctg caatgcggca attcagtggt tcgtccaatc catgtcagac ccgtctgttg
     5101 ccttcctaat aaggcacgat cgtaccacct tacttccacc aatcggcatg cacggtgctt
     5161 tttctctcct tgtaaggcat gttgctaact catcgttacc atgttgcaag actacaagag
     5221 tattgcataa gactacatta agcttgcagc tccagctttt gttcccttta gtgagggtta
     5281 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5341 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5401 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5461 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5521 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5581 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5641 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5701 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5761 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5821 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5881 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5941 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6001 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6061 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6121 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6181 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6241 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6301 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6361 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6421 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6481 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6541 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6601 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6661 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6721 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6781 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6841 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6901 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6961 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7021 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7081 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7141 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7201 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7261 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7321 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7381 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7441 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     7501 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     7561 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     7621 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     7681 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     7741 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     7801 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     7861 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     7921 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     7981 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     8041 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
     8101 aattgggtac gtccctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     8161 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     8221 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggatagctag
     8281 agccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     8341 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     8401 gcaataaaca agttcctctc actctctgat attcatttct ttgcaagtta taaatactga
     8461 ataataagat gacatgaact actactgcta gagattttcc acactgacta aaagggtctg
     8521 agggatctct agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa
     8581 ggcaagcttt attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct
     8641 cagatctggt ctaaccagag agacccagta caagcaaaaa gcagatcttg tcttcgttgg
     8701 gagtgagcta gcccttccag tccccccttt tcttttaaaa agtggctaag atctacagct
     8761 gccttgtaag tcattggtct taaaggtacc cggtatcgat gcggggaggc ggcccaaagg
     8821 gagatccgac tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc
     8881 aggccgcggg aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc
     8941 ccgcgcagaa tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc
     9001 gacaacacca cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg
     9061 caggcggcga tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg
     9121 agctgacagg tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac
     9181 accacgccac gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat
     9241 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     9301 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     9361 tcacaaattt tgtaatccag aggttgatta tcgataagct tgatatcgaa ttgtacctag
     9421 tggaaccgga acccttaaac atgtataact tcgtataatg tatgctatac gaagttatta
     9481 ggtccctcga cgaattccta cttgtacact attcctttgc cctcggacga gtgctggggc
     9541 gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc
     9601 tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc
     9661 gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt
     9721 caagaccaat gcggagcata tacgcccgga gccgcggcga tcctgcaagc tccggatgcc
     9781 tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga
     9841 tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg
     9901 ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc
     9961 ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg
    10021 cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc
    10081 atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc
    10141 cgctcgtctg gctaagatcg gccgcagcga tcgcatccat ggcctccgcg accggctgca
    10201 gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg
    10261 agatgcaata ggtcaggctc tcgctgaatt ccccaatgtc aagcact
//
