LOCUS       sequence-361674-        9057 bp    DNA     circular SYN 14-APR-2025
DEFINITION  3rd generation, Inducible lentiviral plasmid for expression of human
            wildtype SPTLC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9057)
  AUTHORS   Cordes T, Kuna RS, McGregor GH, Khare SV, Gengatharan J, Muthusamy 
            T, Metallo CM
  TITLE     1-deoxysphingolipid synthesis compromises anchorage-independent 
            growth and plasma membrane endocytosis in cancer cells.
  JOURNAL   J Lipid Res. 2022 Sep 14:100281. doi: 10.1016/j.jlr.2022.100281.
  PUBMED    36115594
REFERENCE   2  (bases 1 to 9057)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Lipid 
            Res. 2022 Sep 14:100281. doi: 10.1016/j.jlr.2022.100281."
COMMENT     Sequence Label: pCW57.1_SPTLC1_WT
FEATURES             Location/Qualifiers
     source          1..9057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    11..29
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    47..65
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    118..136
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    154..172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     219..243
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1692..1709
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1719..2229
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2148..2166
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2239..2835
                     /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(2239..2258)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2735..2755
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2836..2889
                     /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(2890..3507,3508..3513,3514..3549,3550..3552,3553..3588,
                     3589..3591,3592..3627,3628..3636)
                     /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(3676..3696)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4094..4105)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4310..4543
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4615..4736
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4652..4671)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4706..4725
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4776..4911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4911..4982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4983..5051,5052..5843)
                     /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(5201..5220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6014..6602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6503..6522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6756..6773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6890..6911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6926..6931,6932..6949,6950..6956)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6964..6980
                     /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     6969..6991
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6988..7004
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6988..7004
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7023..7043
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7025..7043
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7071..7297
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7298..7478
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7525..7650
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8143..8376
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8561..8605
                     /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    8818..8935
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(8989..9057,1..246)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    8997..9015
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9033..9051
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
       61 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
      121 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      181 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      241 atcgcctgga gaattggcta gcatggcgac cgccacggag cagtgggttc tggtggagat
      301 ggtacaggcg ctttacgagg ctcctgctta ccatcttatt ttggaaggga ttctgatcct
      361 ctggataatc agacttcttt tctctaagac ttacaaatta caagaacgat ctgatcttac
      421 agtcaaggaa aaagaagaac tgattgaaga gtggcaacca gaacctcttg ttcctcctgt
      481 cccaaaagac catcctgctc tcaactacaa catcgtttca ggccctccaa gccacaaaac
      541 tgtggtgaat ggaaaagaat gtataaactt cgcctcattt aattttcttg gattgttgga
      601 taaccctagg gttaaggcag cagctttagc atctctaaag aagtatggcg tggggacttg
      661 tggacccaga ggattttatg gcacatttga tgttcatttg gatttggaag accgcctggc
      721 aaaatttatg aagacagaag aagccattat atactcatat ggatttgcca ccatagccag
      781 tgctattcct gcttactcta aaagagggga cattgttttt gtagatagag ctgcctgctt
      841 tgctattcag aaaggattac aggcatcccg tagtgacatt aagttattta agcataatga
      901 catggctgac ctcgagcgac tactaaaaga acaagagatc gaagatcaaa agaatcctcg
      961 caaggctcgt gtaactcggc gtttcattgt agtagaagga ttgtatatga atactggaac
     1021 tatttgtcct cttccagaat tggttaagtt aaaatacaaa tacaaagcaa gaatcttcct
     1081 ggaggaaagc ctttcatttg gagtcctagg agagcatggc cgaggagtca ctgaacacta
     1141 tggaatcaat attgatgata ttgatcttat cagtgccaac atggagaatg cacttgcttc
     1201 tattggaggt ttctgctgtg gcaggtcttt tgtaattgac catcagcgac tttccggcca
     1261 gggatactgc ttttcagctt cgttacctcc cctgttagct gctgcagcaa ttgaggccct
     1321 caacatcatg gaagagaatc caggtatttt tgcagtgttg aaggaaaagt gcggacaaat
     1381 tcataaagct ttacaaggca tttctggatt aaaagtggtg ggggagtccc tttctccagc
     1441 ctttcaccta caactggaag agagcactgg gtctcgcgag caagatgtca gactgcttca
     1501 ggaaattgta gatcaatgca tgaacagaag tattgcatta actcaggcgc gctacttgga
     1561 gaaagaagag aagtgtctcc ctcctcccag cattcgggtt gtggtcacgg tggaacaaac
     1621 agaggaagaa ctggagagag ctgcgtccac catcaaggag gtagcccagg ccgtcctgct
     1681 ctagaccggt ccaccaccac caccaccact aaggatccgg ggttggggtt gcgccttttc
     1741 caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc
     1801 agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc
     1861 ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct aagtcgggaa
     1921 ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg tctcactagt
     1981 accctcgcag acggacagcg ccagggagca atggcagcgc gccgaccgcg atgggctgtg
     2041 gccaatagcg gctgctcagc agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg
     2101 gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc
     2161 tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat caccgacctc
     2221 tctccccagc aattcaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     2281 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     2341 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     2401 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     2461 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     2521 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     2581 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     2641 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     2701 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     2761 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     2821 cgcaagcccg gtgccgaagg tagaggttct ctcctcactt gtggtgatgt tgaagaaaac
     2881 cctggtccaa tgtctagact ggacaagagc aaagtcataa acggagctct ggaattactc
     2941 aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     3001 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
     3061 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     3121 tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct
     3181 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     3241 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     3301 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
     3361 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     3421 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     3481 gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga ttttgactta
     3541 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     3601 cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcg atccaagata
     3661 tcgtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3721 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3781 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3841 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3901 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3961 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     4021 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4081 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4141 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4201 cgcctccccg cctgtttcgc ctcggcgtcc ggactagagg tacctttaag accaatgact
     4261 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     4321 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     4381 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     4441 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     4501 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     4561 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     4621 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4681 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4741 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4801 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4861 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctttcaaata
     4921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4981 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5041 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5101 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5161 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5221 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5281 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5341 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5401 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5461 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5521 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5581 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5641 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5701 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5761 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5821 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5881 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5941 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6001 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6061 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6781 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6841 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6901 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6961 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     7021 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
     7081 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
     7141 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
     7201 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
     7261 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
     7321 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7381 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7441 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     7501 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     7561 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     7621 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     7681 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     7741 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     7801 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     7861 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     7921 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     7981 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     8041 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     8101 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     8161 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     8221 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     8281 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     8341 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8401 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8461 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8521 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8581 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8641 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     8701 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     8761 cccaaccccg aggggacaat tctcgacctc gagacaaatg gcagtattca tccacaattt
     8821 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     8881 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     8941 ttattacagg gacagcagag atccactttg gccgcgaatc gatatgtcga gtttactccc
     9001 tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag aacgatg
//
