LOCUS       Exported               10222 bp ds-DNA     circular SYN 03-JAN-2023
DEFINITION  3rd generation, Inducible lentiviral expression, TRE-gateway; 
            PGK-rtTA-2A-blasti.
ACCESSION   .
VERSION     .
KEYWORDS    pCW57.1_V5_TurboID_FLAG_Nt
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10222)
  TITLE     Superti-Furga RESOLUTE modified pLIX plasmids
REFERENCE   2  (bases 1 to 10222)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 3, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10222
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    36..54
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    71..89
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    107..125
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    143..161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    178..196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    214..232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     279..303
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             373..1329
                     /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"
     CDS             1360..1383
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1396..1520
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1545..1575
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1629..2288
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1695..1714)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2630..2935
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2814..2833
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2976..3100)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3118..3135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        3145..3655
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3574..3592
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3665..4060
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4061..4114
                     /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             4115..4861
                     /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(4901..4921)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5319..5330)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5535..5768
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5840..5961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5877..5896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5931..5950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6001..6136
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6063..6082
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6136..6207
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6208..7068
                     /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(6426..6445)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7239..7827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7728..7747
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7981..7998
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8115..8136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8151..8181
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8189..8205
                     /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     8194..8216
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8213..8229
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8213..8229
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8248..8268
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8250..8268
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8296..8522
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8523..8703
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8750..8875
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9368..9601
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9786..9830
                     /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    10043..10160
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        10214..306
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    10222..18
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
       61 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
      121 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
      181 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      241 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      301 atcgcctgga gaattggcta gcatcaaacc atgggaccca tccccaaccc cctgctgggc
      361 ctggacagca ccaaagacaa tactgtgcct ctgaagctga tcgctctcct ggctaatggc
      421 gagttccata gtggcgaaca gctgggagaa accctgggca tgtccagggc cgctatcaac
      481 aagcacattc agactctgcg cgactggggc gtggacgtgt tcaccgtgcc cggaaagggc
      541 tactctctgc ccgagcctat cccgctgctg aacgctaaac agattctggg acagctggac
      601 ggcgggagcg tggcagtcct gcctgtggtc gactccacca atcagtacct gctggatcga
      661 atcggcgagc tgaagagtgg ggatgcttgc attgcagaat atcagcaggc agggagagga
      721 agcagaggga ggaaatggtt ctctcctttt ggagctaacc tgtacctgag tatgttttgg
      781 cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg tcatcggaat tgtcatggca
      841 gaagcgctgc gaaagctggg agcagacaag gtgcgagtca aatggcccaa tgacctgtat
      901 ctgcaggata gaaagctggc aggcatcctg gtggagctgg ccggaataac aggcgatgct
      961 gcacagatcg tcattggcgc cgggattaac gtggctatga ggcgcgtgga ggaaagcgtg
     1021 gtcaatcagg gctggatcac actgcaggaa gcagggatta acctggacag gaatactctg
     1081 gccgctacgc tgatccgaga gctgcgggca gccctggaac tgttcgagca ggaaggcctg
     1141 gctccatatc tgccacggtg ggagaagctg gataacttca tcaatagacc cgtgaagctg
     1201 atcattgggg acaaagagat tttcgggatt agccggggga ttgataaaca gggagccctg
     1261 ctgctggaac aggacggagt tatcaaaccc tggatgggcg gagaaatcag tctgcggtct
     1321 gccgaaaagg ggggatctgg ccccggcggc ggcgcgccag actacaaaga cgatgacgat
     1381 aaactaggcg cgcccacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata
     1441 aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca
     1501 caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt
     1561 ccggctcgta taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg
     1621 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     1681 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     1741 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     1801 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
     1861 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     1921 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     1981 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     2041 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     2101 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     2161 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     2221 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     2281 gggcgtaaag atctggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg
     2341 ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag
     2401 gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa
     2461 ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt
     2521 cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg
     2581 tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa
     2641 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     2701 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     2761 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     2821 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     2881 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     2941 aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt
     3001 tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat
     3061 cattttacgt ttctcgttca gctttcttgt acaaagtggt ttagtaatga accggtccac
     3121 caccaccacc accactaagg atccggggtt ggggttgcgc cttttccaag gcagccctgg
     3181 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
     3241 tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
     3301 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
     3361 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
     3421 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
     3481 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
     3541 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
     3601 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccagcaatt
     3661 caccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
     3721 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
     3781 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
     3841 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
     3901 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
     3961 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
     4021 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc gaaggtagag gttctctcct
     4081 cacttgtggt gatgttgaag aaaaccctgg tccaatgtct agactggaca agagcaaagt
     4141 cataaacgga gctctggaat tactcaatgg tgtcggtatc gaaggcctga cgacaaggaa
     4201 actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga agaacaagcg
     4261 ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc acttctgccc
     4321 cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat accgctgtgc
     4381 tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa cagagaaaca
     4441 gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct ccctggagaa
     4501 cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat tggaggaaca
     4561 ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta tgcccccact
     4621 tctgagacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct tccttttcgg
     4681 cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg gcgggccgac
     4741 cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg acgactttga
     4801 ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc tccccgggta
     4861 actaagtaag gatcgatcca agatatcgta ttcttaacta tgttgctcct tttacgctat
     4921 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     4981 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     5041 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     5101 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     5161 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     5221 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     5281 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     5341 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     5401 agacgagtcg gatctccctt tgggccgcct ccccgcctgt ttcgcctcgg cgtccggact
     5461 agaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     5521 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     5581 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     5641 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     5701 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     5761 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     5821 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     5881 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5941 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     6001 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6061 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     6121 ggcttttttg gaggcctttc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6181 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6241 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6301 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6361 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6421 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6481 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6541 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6601 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6661 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6721 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6781 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6841 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6901 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6961 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     7021 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7081 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7141 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7201 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7261 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7321 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7381 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7441 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7501 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7561 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7621 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7681 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7741 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7801 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7861 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7921 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7981 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     8041 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     8101 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     8161 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     8221 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     8281 gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga
     8341 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag
     8401 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac
     8461 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa
     8521 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     8581 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     8641 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     8701 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     8761 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     8821 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     8881 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     8941 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     9001 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     9061 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     9121 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     9181 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     9241 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     9301 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     9361 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     9421 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     9481 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     9541 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     9601 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     9661 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     9721 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     9781 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     9841 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     9901 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     9961 caccattatc gtttcagacc cacctcccaa ccccgagggg acaattctcg acctcgagac
    10021 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
    10081 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
    10141 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca ctttggccgc
    10201 gaatcgatat gtcgagttta ct
//
