V5-GFP-TurboID-KDEL_pLX208

LOCUS       V5-GFP-TurboID-KDEL_pLX208              10080 bp ds-DNA     circular SYN 23-JUN-2023
DEFINITION  expresses GFP-TurboID in the mammalian ER lumen, lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    V5-GFP-TurboID-KDEL_pLX208
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10080)
  AUTHORS   Xu WQ, Cheah JS, Xu C, Messing J, Freibaum BD, Boeynaems S, Taylor 
            JP, Udeshi ND, Carr SA, Ting AY
  TITLE     Dynamic mapping of proteome trafficking within and between living 
            cells by TransitID.
  JOURNAL   bioRxiv. 2023 Feb 8:2023.02.07.527548. doi: 
            10.1101/2023.02.07.527548. Preprint.
  PUBMED    36798302
REFERENCE   2  (bases 1 to 10080)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 23, 2023 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10080
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        913..943
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2130..2363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2548..2592
                     /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        2767..3267
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3186..3204
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3279..4298
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     misc_feature    4355..4472
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        4588..4891
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4892..5095
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5046..5066
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     sig_peptide     5121..5183
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
     CDS             5184..5225
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5232..5948
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             5232..5948
                     /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(5277..5298)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5538..5557)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5885..5906
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5967..6923
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     misc_feature    6990..7578
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7043..7063)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7461..7472)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7650..7883
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7961..8082
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7998..8017)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8052..8071
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8122..8257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8184..8203
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(8277..8296)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8278..8296)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8306..8323)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8306..8322)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8315..8337)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8464..8919
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8551..8570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8761..8782
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8945..9049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9050..9910
                     /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(9268..9287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggctgagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2761 ggacccttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
     2821 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
     2881 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
     2941 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
     3001 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
     3061 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
     3121 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
     3181 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
     3241 gaccgaatca ccgacctctc tccccagggg gtaccaccat gaaaaagcct gaactcaccg
     3301 ctacctctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     3361 tctccgaggg cgaagaatct cgggctttca gcttcgatgt gggagggcgt ggatatgtcc
     3421 tgcgggtgaa tagctgcgcc gatggtttct acaaagatcg ctatgtttat cggcactttg
     3481 catccgccgc tctccctatt cccgaagtgc ttgacattgg ggagttcagc gagagcctga
     3541 cctattgcat ctcccgccgt gcacagggtg tcaccttgca agacctgcct gaaaccgaac
     3601 tgcccgctgt tctccagccc gtcgccgagg ccatggatgc catcgctgcc gccgatctta
     3661 gccagaccag cgggttcggc ccattcggac ctcaaggaat cggtcaatac actacatggc
     3721 gcgatttcat ctgcgctatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     3781 acaccgtcag tgcctccgtc gcccaggctc tcgatgagct gatgctttgg gccgaggact
     3841 gccccgaagt ccggcacctc gtgcacgccg atttcggctc caacaatgtc ctgaccgaca
     3901 atggccgcat aacagccgtc attgactgga gcgaggccat gttcggggat tcccaatacg
     3961 aggtcgccaa catcttcttc tggaggccct ggttggcttg tatggagcag cagacccgct
     4021 acttcgagcg gaggcatccc gagcttgcag gatctcctcg gctccgggct tatatgctcc
     4081 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     4141 gggctcaggg tcgctgcgac gcaatcgtcc ggtccggagc cgggactgtc gggcgtacac
     4201 aaatcgcccg cagaagcgct gccgtctgga ccgatggctg tgtggaagtg ctcgccgata
     4261 gtggaaacag acgccccagc actcgtccta gggcaaagga tctgcagtaa tgagaattct
     4321 agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     4381 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     4441 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     4501 cactttggcg ccggctcagg gggcccgggg ttattaatag taatcaatta cggggtcatt
     4561 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4621 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4681 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     4741 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     4801 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     4861 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     4921 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4981 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     5041 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     5101 gctaactgtc gggatccacc atggagacag acacactcct gctatgggta ctgctgctct
     5161 gggttccagg ttccactggt gacggcaagc ccatccccaa ccccctgctg ggcctggaca
     5221 gcaccggatc catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     5281 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     5341 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     5401 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     5461 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     5521 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     5581 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     5641 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     5701 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     5761 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     5821 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     5881 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     5941 tgtacaaggg gggaggcggg ggtagtaaag acaatactgt gcctctgaag ctgatcgctc
     6001 tcctggctaa tggcgagttc catagtggcg aacagctggg agaaaccctg ggcatgtcca
     6061 gggccgctat caacaagcac attcagactc tgcgcgactg gggcgtggac gtgttcaccg
     6121 tgcccggaaa gggctactct ctgcccgagc ctatcccgct gctgaacgct aaacagattc
     6181 tgggacagct ggacggcggg agcgtggcag tcctgcctgt ggtcgactcc accaatcagt
     6241 acctgctgga tcgaatcggc gagctgaaga gtggggatgc ttgcattgca gaatatcagc
     6301 aggcagggag aggaagcaga gggaggaaat ggttctctcc ttttggagct aacctgtacc
     6361 tgagtatgtt ttggcgcctg aagcggggac cagcagcaat cggcctgggc ccggtcatcg
     6421 gaattgtcat ggcagaagcg ctgcgaaagc tgggagcaga caaggtgcga gtcaaatggc
     6481 ccaatgacct gtatctgcag gatagaaagc tggcaggcat cctggtggag ctggccggaa
     6541 taacaggcga tgctgcacag atcgtcattg gcgccgggat taacgtggct atgaggcgcg
     6601 tggaggaaag cgtggtcaat cagggctgga tcacactgca ggaagcaggg attaacctgg
     6661 acaggaatac tctggccgct acgctgatcc gagagctgcg ggcagccctg gaactgttcg
     6721 agcaggaagg cctggctcca tatctgccac ggtgggagaa gctggataac ttcatcaata
     6781 gacccgtgaa gctgatcatt ggggacaaag agattttcgg gattagccgg gggattgata
     6841 aacagggagc cctgctgctg gaacaggacg gagttatcaa accctggatg ggcggagaaa
     6901 tcagtctgcg gtctgccgaa aagggtggag gcgggtctgg aggcgggggt agtaccggta
     6961 aggacgagct gtaaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa
     7021 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     7081 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     7141 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     7201 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     7261 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     7321 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     7381 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     7441 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     7501 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     7561 tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct
     7621 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     7681 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7741 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7801 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7861 gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta
     7921 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     7981 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     8041 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     8101 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     8161 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     8221 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     8281 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     8341 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     8401 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     8461 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     8521 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     8581 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     8641 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     8701 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     8761 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     8821 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     8881 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     8941 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     9001 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     9061 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     9121 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     9181 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     9241 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     9301 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     9361 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     9421 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     9481 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     9541 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     9601 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     9661 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     9721 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     9781 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     9841 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     9901 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     9961 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
    10021 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//