LOCUS       sequence-380713-       10181 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 10181)
  TITLE     Superti-Furga RESOLUTE modified pLIX plasmids
REFERENCE   2  (bases 1 to 10181)
  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_Ct
FEATURES             Location/Qualifiers
     source          1..10181
                     /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"
     protein_bind    383..507
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(532..537,538..555,556..562)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             616..1275
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(682..701)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1617..1922
                     /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     1801..1820
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1963..2087)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2098..2121
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2152..2499,2500..2502,2503..3111)
                     /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"
     promoter        3160..3670
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3589..3607
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3680..4075
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4076..4129
                     /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(4130..4747,4748..4753,4754..4789,4790..4792,4793..4828,
                     4829..4831,4832..4867,4868..4876)
                     /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(4916..4936)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5334..5345)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5550..5783
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5855..5976
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5892..5911)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5946..5965
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6016..6151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6151..6222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6223..6291,6292..7083)
                     /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(6441..6460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7254..7842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7743..7762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7996..8013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8130..8151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8166..8171,8172..8189,8190..8196)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8204..8220
                     /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     8209..8231
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8228..8244
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8228..8244
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8263..8283
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8265..8283
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8311..8537
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8538..8718
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8765..8890
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9383..9616
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9801..9845
                     /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    10058..10175
                     /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 caacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa
      421 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
      481 taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt
      541 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga
      601 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
      661 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
      721 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
      781 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
      841 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
      901 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
      961 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     1021 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     1081 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     1141 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     1201 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     1261 cagggcgggg cgtaaagatc tggatccggc ttactaaaag ccagataaca gtatgcgtat
     1321 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
     1381 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
     1441 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
     1501 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
     1561 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
     1621 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     1681 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
     1741 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     1801 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
     1861 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     1921 aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt
     1981 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     2041 tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggttga ttctagagac
     2101 tacaaagacg atgacgataa accagcgggc ggcggccccg gctctggagg gaaggacaac
     2161 accgtgcccc tgaagctgat cgccctgctg gccaacggcg agttccacag cggcgagcag
     2221 ctgggcgaga ccctgggcat gagccgggcc gccatcaaca agcacatcca gaccctgcgg
     2281 gactggggcg tggacgtgtt caccgtgccc ggcaagggct acagcctgcc cgagcccatc
     2341 cagctgctga acgccaagca gatcctgggc cagctggacg gcggcagcgt ggccgtgctg
     2401 cccgtgatcg acagcaccaa ccagtacctg ctggaccgga tcggcgagct gaagagcggc
     2461 gacgcctgca tcgccgagta ccagcaggcc ggccggggcg gccggggccg gaagtggttc
     2521 agccccttcg gcgccaacct gtacctgagc atgttctggc ggctggagca gggccccgcc
     2581 gccgccatcg gcctgagcct ggtgatcggc atcgtgatgg ccgaggtgct gcggaagctg
     2641 ggcgccgaca aggtgcgggt gaagtggccc aacgacctgt acctgcagga ccggaagctg
     2701 gccggcatcc tggtggagct gaccggcaag accggcgacg ccgcccagat cgtgatcggc
     2761 gccggcatca acatggccat gcggcgggtg gaggagagcg tggtgaacca gggctggatc
     2821 accctgcagg aggccggcat caacctggac cggaacaccc tggccgccat gctgatccgg
     2881 gagctgcggg ccgccctgga gctgttcgag caggagggcc tggcccccta cctgagccgg
     2941 tgggagaagc tggacaactt catcaaccgg cccgtgaagc tgatcatcgg cgacaaggag
     3001 atcttcggca tcagccgggg catcgacaag cagggcgccc tgctgctgga gcaggacggc
     3061 atcatcaagc cctggatggg cggcgagatc agcctgcgga gcgccgagaa gggacccatc
     3121 cccaaccccc tgctgggcct ggacagcacc taaggatccg gggttggggt tgcgcctttt
     3181 ccaaggcagc cctgggtttg cgcagggacg cggctgctct gggcgtggtt ccgggaaacg
     3241 cagcggcgcc gaccctgggt ctcgcacatt cttcacgtcc gttcgcagcg tcacccggat
     3301 cttcgccgct acccttgtgg gccccccggc gacgcttcct gctccgcccc taagtcggga
     3361 aggttccttg cggttcgcgg cgtgccggac gtgacaaacg gaagccgcac gtctcactag
     3421 taccctcgca gacggacagc gccagggagc aatggcagcg cgccgaccgc gatgggctgt
     3481 ggccaatagc ggctgctcag cagggcgcgc cgagagcagc ggccgggaag gggcggtgcg
     3541 ggaggcgggg tgtggggcgg tagtgtgggc cctgttcctg cccgcgcggt gttccgcatt
     3601 ctgcaagcct ccggagcgca cgtcggcagt cggctccctc gttgaccgaa tcaccgacct
     3661 ctctccccag caattcacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     3721 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     3781 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     3841 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     3901 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     3961 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     4021 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggcgaagg
     4081 tagaggttct ctcctcactt gtggtgatgt tgaagaaaac cctggtccaa tgtctagact
     4141 ggacaagagc aaagtcataa acggagctct ggaattactc aatggtgtcg gtatcgaagg
     4201 cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc tgtactggca
     4261 cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc gagatgctgg acaggcatca
     4321 tacccacttc tgccccctgg aaggcgagtc atggcaagac tttctgcgga acaacgccaa
     4381 gtcataccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc tcggcacccg
     4441 cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg
     4501 cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta cactgggctg
     4561 cgtattggag gaacaggagc atcaagtagc aaaagaggaa agagagacac ctaccaccga
     4621 ttctatgccc ccacttctga gacaagcaat tgagctgttc gaccggcagg gagccgaacc
     4681 tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc taaagtgcga
     4741 aagcggcggg ccgaccgacg cccttgacga ttttgactta gacatgctcc cagccgatgc
     4801 ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt ttgaccttga
     4861 catgctcccc gggtaactaa gtaaggatcg atccaagata tcgtattctt aactatgttg
     4921 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4981 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5041 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5101 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5161 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5221 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     5281 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5341 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5401 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctgtttcgc
     5461 ctcggcgtcc ggactagagg tacctttaag accaatgact tacaaggcag ctgtagatct
     5521 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     5581 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     5641 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     5701 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     5761 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     5821 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     5881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5941 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     6001 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6061 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6121 cagaagtagt gaggaggctt ttttggaggc ctttcaaata tgtatccgct catgagacaa
     6181 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6241 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6301 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6361 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6421 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6481 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6541 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6601 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6661 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6721 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6781 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6841 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6901 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6961 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7021 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7081 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7141 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7201 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7261 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7321 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7381 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7621 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7921 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7981 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     8041 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     8101 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     8161 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     8221 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     8281 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
     8341 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     8401 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     8461 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     8521 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     8581 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     8641 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     8701 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     8761 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     8821 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     8881 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     8941 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     9001 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     9061 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     9121 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     9181 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     9241 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     9301 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     9361 gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa
     9421 gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta
     9481 tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac
     9541 tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa
     9601 aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg
     9661 tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct
     9721 ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag
     9781 aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa
     9841 gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga
     9901 tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag
     9961 ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg aggggacaat
    10021 tctcgacctc gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
    10081 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
    10141 aattacaaaa acaaattaca aaaattcaaa attttcgggt t
//
