LOCUS       sequence-380711-       10225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation, Inducible lentiviral expression, TRE-gateway; 
            PGK-rtTA-2A-blasti.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10225)
  TITLE     Superti-Furga RESOLUTE modified pLIX plasmids
REFERENCE   2  (bases 1 to 10225)
  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"
COMMENT     Sequence Label: pCW57.1_V5_BirA*_FLAG_Nt
FEATURES             Location/Qualifiers
     source          1..10225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..362
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    56..74
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    92..110
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    127..145
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    163..181
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    199..217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    234..252
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    270..288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     335..359
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(429..776,777..779,780..1388)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             1419..1442
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1455..1579
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1604..1609,1610..1627,1628..1634)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1688..2347
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1754..1773)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2689..2994
                     /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     2873..2892
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3035..3159)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3177..3194
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        3204..3714
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3633..3651
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3724..4119
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4120..4173
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4174..4791,4792..4797,4798..4833,4834..4836,4837..4872,
                     4873..4875,4876..4911,4912..4920)
                     /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(4960..4980)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5378..5389)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5594..5827
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5899..6020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5936..5955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5990..6009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6060..6195
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6195..6266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6267..6335,6336..7127)
                     /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(6485..6504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7298..7886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7787..7806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8040..8057
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8174..8195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8210..8215,8216..8233,8234..8240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8248..8264
                     /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     8253..8275
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8272..8288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8272..8288
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8307..8327
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8309..8327
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8355..8581
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8582..8762
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8809..8934
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9427..9660
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9845..9889
                     /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    10102..10219
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tattacaggg acagcagaga tccactttgg ccgcgaatcg atatgtcgag tttactccct
       61 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgatgtcga
      121 gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
      181 aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt ttatccctat
      241 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag
      301 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
      361 cctggagaat tggctagcat caaaccatgg gacccatccc caaccccctg ctgggcctgg
      421 acagcaccaa ggacaacacc gtgcccctga agctgatcgc cctgctggcc aacggcgagt
      481 tccacagcgg cgagcagctg ggcgagaccc tgggcatgag ccgggccgcc atcaacaagc
      541 acatccagac cctgcgggac tggggcgtgg acgtgttcac cgtgcccggc aagggctaca
      601 gcctgcccga gcccatccag ctgctgaacg ccaagcagat cctgggccag ctggacggcg
      661 gcagcgtggc cgtgctgccc gtgatcgaca gcaccaacca gtacctgctg gaccggatcg
      721 gcgagctgaa gagcggcgac gcctgcatcg ccgagtacca gcaggccggc cggggcggcc
      781 ggggccggaa gtggttcagc cccttcggcg ccaacctgta cctgagcatg ttctggcggc
      841 tggagcaggg ccccgccgcc gccatcggcc tgagcctggt gatcggcatc gtgatggccg
      901 aggtgctgcg gaagctgggc gccgacaagg tgcgggtgaa gtggcccaac gacctgtacc
      961 tgcaggaccg gaagctggcc ggcatcctgg tggagctgac cggcaagacc ggcgacgccg
     1021 cccagatcgt gatcggcgcc ggcatcaaca tggccatgcg gcgggtggag gagagcgtgg
     1081 tgaaccaggg ctggatcacc ctgcaggagg ccggcatcaa cctggaccgg aacaccctgg
     1141 ccgccatgct gatccgggag ctgcgggccg ccctggagct gttcgagcag gagggcctgg
     1201 ccccctacct gagccggtgg gagaagctgg acaacttcat caaccggccc gtgaagctga
     1261 tcatcggcga caaggagatc ttcggcatca gccggggcat cgacaagcag ggcgccctgc
     1321 tgctggagca ggacggcatc atcaagccct ggatgggcgg cgagatcagc ctgcggagcg
     1381 ccgagaaggg gggatctggc cccggcggcg gcgcgccaga ctacaaagac gatgacgata
     1441 aactaggcgc gcccacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
     1501 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     1561 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
     1621 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
     1681 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     1741 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1801 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1861 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     1921 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1981 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     2041 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     2101 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     2161 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     2221 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     2281 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     2341 ggcgtaaaga tctggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     2401 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     2461 tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     2521 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     2581 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     2641 ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag
     2701 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     2761 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     2821 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     2881 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     2941 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     3001 ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt
     3061 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     3121 attttacgtt tctcgttcag ctttcttgta caaagtggtt tagtaatgaa ccggtccacc
     3181 accaccacca ccactaagga tccggggttg gggttgcgcc ttttccaagg cagccctggg
     3241 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
     3301 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
     3361 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
     3421 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
     3481 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
     3541 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
     3601 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
     3661 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagcaattc
     3721 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     3781 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     3841 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     3901 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     3961 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     4021 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     4081 attcgtgaat tgctgccctc tggttatgtg tgggagggcg aaggtagagg ttctctcctc
     4141 acttgtggtg atgttgaaga aaaccctggt ccaatgtcta gactggacaa gagcaaagtc
     4201 ataaacggag ctctggaatt actcaatggt gtcggtatcg aaggcctgac gacaaggaaa
     4261 ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg
     4321 gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc
     4381 ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct
     4441 ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag
     4501 tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac
     4561 gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag
     4621 gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt
     4681 ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc
     4741 ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc
     4801 gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac
     4861 cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa
     4921 ctaagtaagg atcgatccaa gatatcgtat tcttaactat gttgctcctt ttacgctatg
     4981 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     5041 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     5101 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     5161 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     5221 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     5281 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     5341 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     5401 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     5461 gacgagtcgg atctcccttt gggccgcctc cccgcctgtt tcgcctcggc gtccggacta
     5521 gaggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     5581 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     5641 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5701 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5761 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     5821 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     5881 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5941 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6001 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     6061 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     6121 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     6181 gcttttttgg aggcctttca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6241 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6301 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6361 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6421 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6481 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6541 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6601 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6661 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6721 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6781 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6841 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6901 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6961 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7021 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7081 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7141 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7201 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7261 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7321 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7381 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7441 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7501 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7561 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7621 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7681 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7741 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7801 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7861 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7921 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7981 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     8041 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     8101 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     8161 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     8221 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     8281 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     8341 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     8401 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     8461 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     8521 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     8581 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     8641 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     8701 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     8761 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     8821 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     8881 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     8941 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     9001 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     9061 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     9121 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     9181 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     9241 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     9301 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     9361 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     9421 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     9481 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     9541 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     9601 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     9661 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     9721 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     9781 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     9841 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     9901 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     9961 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
    10021 accattatcg tttcagaccc acctcccaac cccgagggga caattctcga cctcgagaca
    10081 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
    10141 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
    10201 tacaaaaatt caaaattttc gggtt
//
