LOCUS       sequence-375119-        9668 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lenti-PL(Puro-Luciferase).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9668)
  AUTHORS   Ye L, Park JJ, Peng L, Yang Q, Chow RD, Dong MB, Lam SZ, Guo J, Tang
            E, Zhang Y, Wang G, Dai X, Du Y, Kim HR, Cao H, Errami Y, Clark P, 
            Bersenev A, Montgomery RR, Chen S
  TITLE     A genome-scale gain-of-function CRISPR screen in CD8 T cells 
            identifies proline metabolism as a means to enhance CAR-T therapy.
  JOURNAL   Cell Metab. 2022 Apr 5;34(4):595-614.e14. doi: 
            10.1016/j.cmet.2022.02.009. Epub 2022 Mar 10.
  PUBMED    35276062
REFERENCE   2  (bases 1 to 9668)
  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"; doi: 
            "10.1016/j.cmet.2022.02.009"; journalName: "Cell Metab"; date: 
            "2022-04-5- 5"; volume: "34"; issue: "4"; pages: "595-614.e14"
COMMENT     Sequence Label: pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE
FEATURES             Location/Qualifiers
     source          1..9668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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    1741..1858
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1997..3175
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2228..3166
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3123..3143
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3188..3784
                     /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(3188..3207)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3684..3704
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3794..3850
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3851..5503
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3917..3935)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    5525..6113
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5578..5598)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5996..6007)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6185..6418
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6496..6617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6533..6552)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6587..6606
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6657..6792
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6812..6831)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6813..6831)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6841..6858)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6841..6857)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6850..6872)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6999..7454
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7086..7105)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7296..7317
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7480..7584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7585..7653,7654..8445)
                     /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(7803..7822)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8616..9204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9105..9124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9358..9375
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9492..9513
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9528..9533,9534..9551,9552..9558)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9566..9582
                     /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     9571..9593
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9590..9606
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9590..9606
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9625..9645
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9627..9645
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccgagacaa atggcagtat tcatccacaa
     1741 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     1801 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     1861 ggtttattac agggacagca gagatccact ttggcgccgg ctcgaggggg cccgggtgca
     1921 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
     1981 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     2041 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     2101 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     2161 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     2221 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
     2281 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
     2341 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
     2401 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
     2461 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
     2521 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
     2581 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
     2641 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
     2701 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
     2761 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
     2821 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     2881 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     2941 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     3001 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     3061 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     3121 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgacgtac
     3181 ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     3241 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     3301 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     3361 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg tctggaccac
     3421 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     3481 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     3541 caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg
     3601 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     3661 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     3721 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     3781 tgccggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa
     3841 tcctggaccg atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct
     3901 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
     3961 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt
     4021 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
     4081 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
     4141 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
     4201 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt
     4261 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga
     4321 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
     4381 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa
     4441 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt
     4501 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
     4561 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
     4621 atgtggattt cgagtcgtct taatgtatag atttgaagag gagctgtttc tgaggagcct
     4681 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa
     4741 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc
     4801 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag
     4861 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
     4921 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
     4981 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat
     5041 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
     5101 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg
     5161 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat
     5221 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc
     5281 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
     5341 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
     5401 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
     5461 gatcctcata aaggccaaga agggcggaaa gatcgccgtg taagaattca cgcgttaagt
     5521 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     5581 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     5641 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     5701 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     5761 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5821 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5881 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     5941 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     6001 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     6061 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact
     6121 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     6181 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     6241 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6301 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6361 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6421 acgtatagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     6481 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6541 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6601 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     6661 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6721 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6781 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     6841 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     6901 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     6961 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     7021 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     7081 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     7141 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     7201 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     7261 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     7321 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     7381 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     7441 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     7501 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     7561 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     7621 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     7681 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     7741 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     7801 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     7861 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     7921 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7981 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     8041 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     8101 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     8161 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     8221 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     8281 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     8341 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     8401 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     8461 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     8521 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     8581 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     8641 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     8701 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     8761 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     8821 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8881 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8941 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     9001 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     9061 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     9121 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     9181 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     9241 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     9301 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     9361 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     9421 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     9481 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     9541 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     9601 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag
     9661 ctgcaagc
//
