LOCUS       sequence-424203-        8019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For lentiviral expression of shRNA under a U6 promoter, accompanied 
            by the expression of the affinity sorting tag GST-EGFP-GPIBY55 
            driven by an hPGK promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8019)
  AUTHORS   Ma S, Yang L, Zuo Q, Huang Q
  TITLE     GPI-anchored glutathione S-transferase as marker allows affinity 
            sorting of transfection-positive cells.
  JOURNAL   Front Mol Biosci. 2022 Sep 29;9:1016090. doi: 
            10.3389/fmolb.2022.1016090. eCollection 2022.
  PUBMED    36250010
REFERENCE   2  (bases 1 to 8019)
  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"; journalName: "Front Mol 
            Biosci."; date: "2022-09-29"; volume: "9:1016090. doi"; pages: " 
            10.3389/fmolb.2022.1016090. eCollection 2022"
COMMENT     Sequence Label: pLKO.1-GST-EGFP-GPIBY55
FEATURES             Location/Qualifiers
     source          1..8019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..511
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     430..448
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             619..1275
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MGSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1233..1255
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1282..1299
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      1306..1315
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1312..1314,1315..1317,1318..2028)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1312..1314,1315..1317,1318..2028)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1357..1378)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1618..1637)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1965..1986
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2229..2462
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2534..2655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2571..2590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2625..2644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2695..2830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2850..2869)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2851..2869)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2879..2896)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2879..2895)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2888..2910)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3037..3492
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3124..3143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3334..3355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3518..3622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3623..3691,3692..4483)
                     /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(3841..3860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4654..5242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5143..5162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5396..5413
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5530..5551
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5566..5571,5572..5589,5590..5596)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5604..5620
                     /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     5609..5631
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5628..5644
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5628..5644
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5663..5683
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5665..5683
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5711..5937
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5938..6118
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6165..6290
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6783..7016
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7201..7245
                     /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"
     promoter        7543..7791
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7543..7563
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7714..7733
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    7854..7971
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc
       61 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc
      121 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc
      181 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac
      241 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc
      301 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag
      361 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct
      421 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct
      481 cgttgaccga atcaccgacc tctctcccca gggggatcca ccatgctgct ggagcctggc
      541 agaggctgct gcgctctggc catcctgctg gccatcgtgg acatccagag cggaggcagc
      601 gcctggagcc accctcagat gggctcccct atactaggtt attggaaaat taagggcctt
      661 gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga gcatttgtat
      721 gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt ggagtttccc
      781 aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc catcatacgt
      841 tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc agagatttca
      901 atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc atatagtaaa
      961 gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct gaaaatgttc
     1021 gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca tcctgacttc
     1081 atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct ggatgcgttc
     1141 ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga taagtacttg
     1201 aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt tggtggtggc
     1261 gaccatcctc caaaatcgga tctggttccg cgtggatccc cggtcgccac catggtgagc
     1321 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     1381 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     1441 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     1501 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     1561 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     1621 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     1681 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     1741 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1801 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1861 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1921 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1981 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc cggccggact
     2041 cagatccggc agcacctgga gttcagccac aacgagggca ccctgagcag cggcttcctc
     2101 caggagaagg tgtgggtgat gctggtgacc agcctggtgg ctctccaggc cctgtagggt
     2161 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     2221 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     2281 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     2341 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     2401 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     2461 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     2521 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2581 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2641 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg
     2701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2761 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2821 tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     2881 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2941 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3001 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     3061 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3121 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3181 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3241 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3301 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3361 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3421 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3481 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3541 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3601 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3661 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3721 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3781 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3841 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3901 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3961 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4021 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4081 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4141 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4201 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4261 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4321 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4381 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4441 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4501 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4561 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4621 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4681 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4741 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4801 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4861 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4921 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4981 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5041 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5101 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5161 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5221 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5281 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5341 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5401 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5461 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5521 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     5581 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     5641 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     5701 gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt
     5761 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg
     5821 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc
     5881 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg
     5941 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6001 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6061 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6121 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     6181 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     6241 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     6301 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     6361 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     6421 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     6481 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     6541 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     6601 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     6661 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     6721 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     6781 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     6841 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     6901 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     6961 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     7021 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     7081 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     7141 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     7201 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     7261 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     7321 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     7381 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     7441 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
     7501 tatcgatcac gagactagcc tcgagcggcc gcccccttca ccgagggcct atttcccatg
     7561 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
     7621 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     7681 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
     7741 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cggtccgcag
     7801 gtatgcacgc gtgaattctc gacctcgaga caaatggcag tattcatcca caattttaaa
     7861 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     7921 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     7981 tacagggaca gcagagatcc actttggccg cggctcgag
//
