LOCUS       sequence-307431-        9428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses codon optimized catalytically inactive PHGDH in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9428)
  AUTHORS   Ngo B, Kim E, Osorio-Vasquez V, Doll S, Bustraan S, Liang RJ, Luengo
            A, Davidson SM, Ali A, Ferraro GB, Fischer GM, Eskandari R, Kang DS,
            Ni J, Plasger A, Rajasekhar VK, Kastenhuber ER, Bacha S, Sriram RK, 
            Stein BD, Bakhoum SF, Snuderl M, Cotzia P, Healey JH, Mainolfi N, 
            Suri V, Friedman A, Manfredi M, Sabatini DM, Jones DR, Yu M, Zhao 
            JJ, Jain RK, Keshari KR, Davies MA, Vander Heiden MG, Hernando E, 
            Mann M, Cantley LC, Pacold ME
  TITLE     Limited Environmental Serine and Glycine Confer Brain Metastasis 
            Sensitivity to PHGDH Inhibition.
  JOURNAL   Cancer Discov. 2020 Sep;10(9):1352-1373. doi: 
            10.1158/2159-8290.CD-19-1228. Epub 2020 Jun 22.
  PUBMED    32571778
REFERENCE   2  (bases 1 to 9428)
  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: 
            "10.1158/2159-8290.CD-19-1228"; journalName: "Cancer Discov"; date: 
            "2020-09"; volume: "10"; issue: "9"; pages: "1352-1373"
COMMENT     Sequence Label: pCW-codon optimized catalytically inactive PHGDH
FEATURES             Location/Qualifiers
     source          1..9428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..362
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    56..74
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    92..110
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    127..145
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    163..181
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    199..217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    234..252
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    270..288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     335..359
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      379..388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2011..2521
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2440..2458
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2531..3322
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2531..3322
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2585..2604)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3195..3214
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             3323..3376
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(3377..3994,3995..4000,4001..4036,4037..4039,4040..4075,
                     4076..4078,4079..4114,4115..4123)
                     /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"
     primer_bind     complement(4163..4183)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4581..4592)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4797..5030
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5102..5223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5139..5158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5193..5212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5263..5398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5398..5469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5470..5538,5539..6330)
                     /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(5688..5707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6501..7089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6990..7009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7243..7260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7377..7398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7413..7418,7419..7436,7437..7443)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7451..7467
                     /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     7456..7478
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7475..7491
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7475..7491
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7510..7530
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7512..7530
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7558..7784
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7785..7965
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8012..8137
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8630..8863
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9048..9092
                     /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    9305..9422
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tattacaggg acagcagaga tccactttgg ccgcgaatcg atatgtcgag tttactccct
       61 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgatgtcga
      121 gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
      181 aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat
      241 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag
      301 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
      361 cctggagaat tggctagcgc caccatggcc tttgcaaatc tcagaaaggt gcttatcagc
      421 gactctcttg acccatgctg ccgcaaaata ctgcaggatg gaggattgca ggtggtagaa
      481 aagcaaaacc tcagtaaaga agagctgata gcagagcttc aagactgcga ggggctgatt
      541 gtaaggagcg cgacgaaggt aactgccgat gttatcaacg ctgcggaaaa actccaggta
      601 gttggtcggg cgggaactgg ggttgataac gtggatcttg aggcagcgac acggaaagga
      661 atccttgtga tgaacacacc taacggcaat agcctttccg ctgcggaact tacctgtggc
      721 atgatcatgt gtctcgcgag gcaaataccc caggctacgg cgagtatgaa agatggtaaa
      781 tgggagcgga agaaattcat gggaactgaa cttaacggta aaactcttgg aatcctggga
      841 ctggggcgga tcggccgcga ggttgcaacc agaatgcagt catttgggat gaagacgatc
      901 ggctataacc ccatcatctc tcctgaggta tccgcatcat tcggggttca acaactgccc
      961 ctggaggaaa tttggcccct ttgcgatttc atcaccgtcc ataccccttt gctcccgagc
     1021 acaactggat tgcttaacga caatacgttt gcgcagtgca aaaagggcgt acgggttgtg
     1081 aactgcgcta aggggggcat tgtagacgag ggcgctttgc tgcgagcttt gcagagtgga
     1141 caatgcgctg gtgccgccct ggatgtattc actgaggaac cgcccagaga cagggcgttg
     1201 gtagaccatg aaaatgtcat tagctgtcca gccttggggg ccagcacgaa agaggcgcag
     1261 agccgatgcg gcgaagagat tgcggtgcaa ttcgtcgaca tggtcaaagg aaaaagtttg
     1321 acgggcgtag tgaacgcgca ggcattgacc tctgcgttct ctccacacac caagccgtgg
     1381 atagggctgg ccgaagccct cggaacgctc atgcgagctt gggcaggtag cccgaaaggt
     1441 actatacagg taatcacaca aggtacttcc ctcaaaaatg ccggtaactg cttgtctcct
     1501 gctgtaatag tcggattgct caaggaagca agtaagcaag cggacgtaaa cctggtaaac
     1561 gccaaacttc tcgtaaaaga agccggactc aatgtcacaa catctcattc cccggcagct
     1621 ccgggggagc agggattcgg cgaatgtctt cttgcggtag ctctggcagg cgctccttat
     1681 caagctgtag gattggttca gggaacaact ccagtcctcc agggactcaa tggtgcggta
     1741 ttcagacccg aggtcccctt gagaagggat ttgccgctcc ttctttttcg cactcagacg
     1801 agcgatcctg ctatgctccc tacaatgatt ggcctgctcg cagaggcagg cgttagattg
     1861 cttagttatc aaacctcact ggtttcagat ggggagacat ggcatgtcat gggtatcagt
     1921 tctctgctcc cgtcactcga agcgtggaag cagcacgtta cggaggcctt ccaattccat
     1981 ttctaaatcg atctcgagaa cgttggatcc ggggttgggg ttgcgccttt tccaaggcag
     2041 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
     2101 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
     2161 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
     2221 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
     2281 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
     2341 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
     2401 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
     2461 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
     2521 gcaattcacc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     2581 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     2641 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     2701 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     2761 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     2821 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     2881 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     2941 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3001 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     3061 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3121 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3181 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3241 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3301 tcgccttctt gacgagttct tcgaaggtag aggttctctc ctcacttgtg gtgatgttga
     3361 agaaaaccct ggtccaatgt ctagactgga caagagcaaa gtcataaacg gagctctgga
     3421 attactcaat ggtgtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg
     3481 agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct
     3541 gccaatcgag atgctggaca ggcatcatac ccacttctgc cccctggaag gcgagtcatg
     3601 gcaagacttt ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga
     3661 cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa
     3721 tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc
     3781 cgccgtgggc cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa
     3841 agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgagac aagcaattga
     3901 gctgttcgac cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg
     3961 tggcctggag aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt
     4021 tgacttagac atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc
     4081 tgacgctctt gacgattttg accttgacat gctccccggg taactaagta aggatcgatc
     4141 caagatatcg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4201 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4261 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4321 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4381 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4441 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4501 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4561 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4621 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4681 tttgggccgc ctccccgcct gtttcgcctc ggcgtccgga ctagaggtac ctttaagacc
     4741 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     4801 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     4861 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     4921 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     4981 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     5041 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     5101 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5161 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5221 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     5281 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5341 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggcctt
     5401 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5461 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5521 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5581 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5641 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5701 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5761 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5821 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5881 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5941 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6001 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6061 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6121 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6181 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6241 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6301 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6361 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6421 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6481 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6541 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6601 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6661 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6721 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6781 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6841 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6901 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6961 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7021 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7081 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7141 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7201 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7261 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7321 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7381 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7441 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7501 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
     7561 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
     7621 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
     7681 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
     7741 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
     7801 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     7861 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     7921 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     7981 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     8041 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     8101 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     8161 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     8221 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     8281 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     8341 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     8401 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     8461 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     8521 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     8581 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     8641 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     8701 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     8761 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     8821 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     8881 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     8941 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     9001 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     9061 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
     9121 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
     9181 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
     9241 cccacctccc aaccccgagg ggacaattct cgacctcgag acaaatggca gtattcatcc
     9301 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     9361 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     9421 ttcgggtt
//
