LOCUS       pHAGE2-EF1a-HA-A       10401 bp DNA     circular SYN 10-FEB-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 10401)
  AUTHORS   Yi Fu
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2023 from SnapGene 7.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10401
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     misc_feature    19..136
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     primer_bind     63..93
                     /label=YF95_cPPT_seq_F
                     /note="31"
                     /note="color: black; sequence: 
                     gaatagtagacataatagcaacagacataca"
     primer_bind     171..255
                     /label=YF180-IsceIphiKO2EF1aMfeI-s
                     /note="188"
                     /note="color: black; sequence: 
                     CAGGGTAATacatgtggaattgtgagcggataacaagctagcGGTAATcagcacacaac
                     agcccattatacgcgcgtataatgggctattatgtgctgCGTGAGGgctagcCGTGAGG
                     CTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGG
                     GAGGGGTCGGC; added: 2021-05-24"
     promoter        184..1362
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(303..323)
                     /label=YF80_EF1a_R
                     /note="21"
                     /note="color: black; sequence: CGGAGCCAGTACACGACATCA"
     intron          415..1353
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
                     /note="color: #------"
     primer_bind     657..679
                     /label=YF74_EF1a-seq_F
                     /note="23"
                     /note="color: black; sequence: CGCTGCTTTCGATAAGTCTCTAG"
     primer_bind     1127..1146
                     /label=YF96_EF1a_F
                     /note="20	pcr xq-neo-18laco-pal-ef1agfp	overlapping pcr"
                     /note="color: black; sequence: CGTCCAGGCACCTCGATTAG"
     primer_bind     complement(1341..1363)
                     /label=YF97_EF1a_R
                     /note="37"
                     /note="color: black; sequence: 
                     TTATCAGATATCtttTCACGACACCTGAAATGGAAGA; added: 2021-05-24"
     primer_bind     1364..1389
                     /label=YF312-MCS-IRES-F
                     /note="35	pcr ireszsg"
                     /note="color: black; sequence: 
                     tgtACGCGTgcggccgcttaattaaGTTTAAACgg; added: 2021-05-24"
     primer_bind     1388..1446
                     /label=YF770-BamHI-ATG-HA-ATAD3A-F
                     /note="color: black; sequence: 
                     taaGGATCCatgTACCCATACGATGTTCCAGATTACGCTTCGTGGCTCTTCGGCATTAA
                     CAA; added: 2022-11-02"
     CDS             1397..1423
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     CDS             1424..3178
                     /codon_start=1
                     /label=ATAD3A CDS
                     /note="color: #ff00ff"
                     /translation="SWLFGINKGPKGEGAGPPPPLPPAQPGAEGGGDRGLGDRPAPKDK
                     WSNFDPTGLERAAKAARELEHSRYAKDALNLAQMQEQTLQLEQQSKLKEYEAAVEQLKS
                     EQIRAQAEERRKTLSEETRQHQARAQYQDKLARQRYEDQLKQQQLLNEENLRKQEESVQ
                     KQEAMRRATVEREMELRHKNEMLRVEAEARARAKAERENADIIREQIRLKAAEHRQTVL
                     ESIRTAGTLFGEGFRAFVTDWDKVTATVAGLTLLAVGVYSAKNATLVAGRFIEARLGKP
                     SLVRETSRITVLEALRHPIQVSRRLLSRPQDALEGVVLSPSLEARVRDIAIATRNTKKN
                     RSLYRNILMYGPPGTGKTLFAKKLALHSGMDYAIMTGGDVAPMGREGVTAMHKLFDWAN
                     TSRRGLLLFVDEADAFLRKRATEKISEDLRATLNAFLYRTGQHSNKFMLVLASNQPEQF
                     DWAINDRINEMVHFDLPGQEERERLVRMYFDKYVLKPATEGKQRLKLAQFDYGRKCSEV
                     ARLTEGMSGREIAQLAVSWQATAYASEDGVLTEAMMDTRVQDAVQQHQQKMCWLKAEGP
                     GRGDEPSPS"
     CDS             1424..2974
                     /codon_start=1
                     /label=delta-Cter
                     /note="color: #ff6600"
                     /translation="SWLFGINKGPKGEGAGPPPPLPPAQPGAEGGGDRGLGDRPAPKDK
                     WSNFDPTGLERAAKAARELEHSRYAKDALNLAQMQEQTLQLEQQSKLKEYEAAVEQLKS
                     EQIRAQAEERRKTLSEETRQHQARAQYQDKLARQRYEDQLKQQQLLNEENLRKQEESVQ
                     KQEAMRRATVEREMELRHKNEMLRVEAEARARAKAERENADIIREQIRLKAAEHRQTVL
                     ESIRTAGTLFGEGFRAFVTDWDKVTATVAGLTLLAVGVYSAKNATLVAGRFIEARLGKP
                     SLVRETSRITVLEALRHPIQVSRRLLSRPQDALEGVVLSPSLEARVRDIAIATRNTKKN
                     RSLYRNILMYGPPGTGKTLFAKKLALHSGMDYAIMTGGDVAPMGREGVTAMHKLFDWAN
                     TSRRGLLLFVDEADAFLRKRATEKISEDLRATLNAFLYRTGQHSNKFMLVLASNQPEQF
                     DWAINDRINEMVHFDLPGQEERERLVRMYFDKYVLKPATEGKQRLKLAQFDYGRKCSEV
                     "
     primer_bind     1435..1454
                     /label=sgATAD3A-Nter
                     /note="color: black; sequence: cggcattaacaagggcccca; added:
                     2022-09-01"
     CDS             1472..1501
                     /codon_start=1
                     /label=Proline rich motif (PRM)
                     /note="color: #993366"
                     /translation="PPPPLPPAQP"
     misc_feature    1585
                     /label=c
                     /note="color: #a6acb3"
     misc_feature    1626..1634
                     /label=sgATAD3A-intron-exon-3'
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1635..1637
                     /label=PAM
                     /note="color: #a6acb3"
     CDS             1673..1765
                     /codon_start=1
                     /product="coiled-coil domain 
                     "
                     /label=CC1
                     /note="color: #993366"
                     /translation="LQLEQQSKLKEYEAAVEQLKSEQIRAQAEER"
     primer_bind     complement(1685..1710)
                     /label=YF779-ATAD3A-N-R
                     /note="color: black; sequence: TCATACTCTTTGAGCTTGGACTGTTG; 
                     added: 2022-11-17"
     primer_bind     complement(1686..1710)
                     /label=Primer 1
                     /note="color: black; sequence: TCATACTCTTTGGGCTTGGACTGTT; 
                     added: 2023-01-12"
     CDS             1697..1699
                     /codon_start=1
                     /product="CC mutant
                     "
                     /label=L93P
                     /note="color: #ff0000"
                     /translation="L"
     primer_bind     complement(1716..1735)
                     /label=sgATAD3A/B
                     /note="67.5*30.2"
                     /note="color: black; sequence: gctcttgagctgctccacgg; added:
                     2022-09-01"
     CDS             1727..1729
                     /codon_start=1
                     /product="cc mutant
                     "
                     /label=L103P
                     /note="color: #ff0000"
                     /translation="L"
     primer_bind     complement(1730..1745)
                     /label=YF802-ATAD3A-L103P-R
                     /note="color: black; sequence: GGATCTGCTCGCTCTTcgG; added: 
                     2023-02-10"
     CDS             1958..2080
                     /codon_start=1
                     /label=CC2
                     /note="color: #993366"
                     /translation="LRHKNEMLRVEAEARARAKAERENADIIREQIRLKAAEHRQ"
     CDS             1997..1999
                     /codon_start=1
                     /product="cc mutant
                     "
                     /label=A193P
                     /note="color: #ff0000"
                     /translation="A"
     CDS             2030..2032
                     /codon_start=1
                     /product="cc mutant
                     "
                     /label=A204P
                     /note="color: #ff0000"
                     /translation="A"
     CDS             2093..2146
                     /codon_start=1
                     /label=TM1
                     /note="color: #993366"
                     /translation="SIRTAGTLFGEGFRAFVT"
     CDS             2159..2212
                     /codon_start=1
                     /label=TM2
                     /note="color: #993366"
                     /translation="VTATVAGLTLLAVGVYSA"
     primer_bind     complement(2201..2235)
                     /label=YF753-ATAD3A-silentmut-R
                     /note="color: black; sequence: 
                     CCGGCcACcAGgGTGGCATTCTTGGCTGAGTAGAC; added: 2022-09-01"
     primer_bind     2213..2232
                     /label=sgATAD3A-synthego
                     /note="specific to ATAD3Anot to ATAD3Bworks wellso mutate 
                     it on plasmid to be resistant to sgRNA"
                     /note="color: black; sequence: aagaatgccacgcttgtcgc; added:
                     2022-09-01"
     primer_bind     2218..2247
                     /label=YF754-ATAD3A-silentmut-F
                     /note="color: black; sequence: 
                     TGCCACcCTgGTgGCCGGCCGCTTCATCGA; added: 2022-09-01"
     CDS             2240..2290
                     /codon_start=1
                     /product="mitochondrial import signal
                     "
                     /label=MIS
                     /note="color: #993366"
                     /translation="FIEARLGKPSLVRETSR"
     CDS             2378..2974
                     /codon_start=1
                     /label=AAA+ ATPase
                     /note="color: #993366"
                     /translation="LSPSLEARVRDIAIATRNTKKNRSLYRNILMYGPPGTGKTLFAKK
                     LALHSGMDYAIMTGGDVAPMGREGVTAMHKLFDWANTSRRGLLLFVDEADAFLRKRATE
                     KISEDLRATLNAFLYRTGQHSNKFMLVLASNQPEQFDWAINDRINEMVHFDLPGQEERE
                     RLVRMYFDKYVLKPATEGKQRLKLAQFDYGRKCSEV"
     CDS             2474..2497
                     /codon_start=1
                     /label=Walker A (ATP binding)
                     /note="color: #993366"
                     /translation="GPPGTGKT"
     CDS             2627..2629
                     /codon_start=1
                     /label=alphafold-potential interact site
                     /note="color: #ffff99"
                     /translation="R"
     CDS             2639..2659
                     /codon_start=1
                     /label=Walker B (ATP binding)
                     /note="color: #993366"
                     /translation="LLFVDEA"
     primer_bind     complement(3162..3188)
                     /label=YF771-BmtI-STOP-ATAD3A-WT-R
                     /note="color: black; sequence: 
                     atcGCTAGCtcaGGATGGGGAGGGCTCGT; added: 2022-11-02"
     primer_bind     complement(3204..3231)
                     /label=YF149_Pointmut_R
                     /note="28"
                     /note="color: black; sequence: 
                     cgacctcgagctctagatacgtaaaatg"
     primer_bind     complement(3229..3252)
                     /label=YF198-GFP110-R
                     /note="24"
                     /note="color: black; sequence: TGGAAAATAACCGGAATTGgtcga"
     primer_bind     3276..3293
                     /label=YF307-AscI-IRES-F
                     /note="color: black; sequence: 
                     ttGGCGCGCCGTGAGGGCCCGGAAACCT"
     misc_feature    3278..3739
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="color: #a6acb3"
     primer_bind     complement(3293..3312)
                     /label=YF633-IRES-R
                     /note="color: black; sequence: TCGTCAAGAAGACAGGGCCA; added:
                     2021-05-20"
     CDS             3744..4769
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /note="color: #ccffcc"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     3747..3766
                     /label=YF192-SphIHygroNoATG_F
                     /note="color: black; sequence: 
                     aatGCATGCAAAAAGCCTGAACTCACCGC"
     primer_bind     complement(4759..4781)
                     /label=YF191-EcoRIHygro-R
                     /note="color: black; sequence: 
                     atcGAATTCGGCATCTACTCTATTCCTTTG"
     primer_bind     complement(4759..4779)
                     /label=YF196-XhoIHygro-R
                     /note="30"
                     /note="color: black; sequence: 
                     atcCTCGAGGGCATCTACTCTATTCCTTTG; added: 2021-05-24"
     misc_feature    4804..5392
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     CDS             complement(5275..5286)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 5274"
                     /translation="IEGR"
     primer_bind     5359..5377
                     /label=YF131_WPREend_F
                     /note="19"
                     /note="color: black; sequence: acgagtcggatctcccttt"
     primer_bind     complement(5393..5416)
                     /label=YF308-Bsu36I-WPRE-R
                     /note="color: black; sequence: 
                     ctaCCTgAGGagtcattggtcttaaaggtaccag"
     LTR             5540..5720
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     promoter        5853..5957
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5958..6818
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5958 .. 6026 / #ccffcc / signal sequence
                        2: 6027 .. 6818 / #ccffcc
                     Cleavage site after base 6026"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6989..7577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(7972..7991)
                     /label=YF370-AAVS1R-F
                     /note="20	use with YF129 for right arm genotyping"
                     /note="color: black; sequence: attaattgcgttgcgctcac"
     protein_bind    7988..8009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        8024..8054
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 8024 .. 8029 / #ffffff / -35
                        2: 8030 .. 8047 / #ffffff
                        3: 8048 .. 8054 / #ffffff / -10"
     protein_bind    8062..8078
                     /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)."
                     /note="color: #31849b"
     primer_bind     8086..8102
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     LTR             8363..8996
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    9043..9168
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    9665..9898
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     CDS             10083..10127
                     /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"
                     /note="color: #cc99b2"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
       61 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      121 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggactagtcg
      181 tgaggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
      241 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
      301 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
      361 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
      421 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
      481 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
      541 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
      601 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
      661 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
      721 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
      781 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
      841 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
      901 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
      961 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
     1021 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
     1081 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     1141 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     1201 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     1261 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     1321 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaagcggccg cttaattaag
     1381 tttaaacgga tccatgtacc catacgatgt tccagattac gcttcgtggc tcttcggcat
     1441 taacaagggc cccaagggtg aaggcgcggg gccgccgccg cctttgccgc ccgcgcagcc
     1501 cggggccgag ggcggcgggg accgcgggtt gggagaccgg ccggcgccca aggacaaatg
     1561 gagcaacttc gaccccaccg gcctggagcg cgccgccaag gcggcgcgcg agctggagca
     1621 ctcgcgttat gccaaggacg ccctgaatct ggcacagatg caggagcaga cgctgcagtt
     1681 ggagcaacag tccaagctca aagagtatga ggccgccgtg gagcagctca agagcgagca
     1741 gatccgggcg caggctgagg agaggaggaa gaccctgagc gaggagaccc ggcagcacca
     1801 ggccagggcc cagtatcaag acaagctggc ccggcagcgc tacgaggacc aactgaagca
     1861 gcagcaactt ctcaatgagg agaatttacg gaagcaggag gagtccgtgc agaagcagga
     1921 agccatgcgg cgagccaccg tggagcggga gatggagctg cggcacaaga atgagatgct
     1981 gcgagtggag gccgaggccc gggcgcgcgc caaggccgag cgggagaatg cagacatcat
     2041 ccgcgagcag atccgcctga aggcggccga gcaccgtcag accgtcttgg agtccatcag
     2101 gacggctggc accttgtttg gggaaggatt ccgtgccttt gtgacagact gggacaaagt
     2161 gacagccacg gtggctgggc tgacgctgct ggctgttggg gtctactcag ccaagaatgc
     2221 cacgcttgtc gccggccgct tcatcgaggc tcggctgggg aagccgtccc tagtgaggga
     2281 gacgtcccgc atcacggtgc ttgaggcgct gcggcacccc atccaggtca gccggcggct
     2341 cctcagtcga ccccaggacg cgctggaggg tgttgtgctc agtcccagcc tggaagcacg
     2401 ggtgcgcgac atcgccatag caacaaggaa caccaagaag aaccgcagcc tgtacaggaa
     2461 catcctgatg tacgggccac caggcaccgg gaagacgctg tttgccaaga aactcgccct
     2521 gcactcaggc atggactacg ccatcatgac aggcggggac gtggccccca tggggcggga
     2581 aggcgtgacc gccatgcaca agctctttga ctgggccaat accagccggc gcggcctcct
     2641 gctctttgtg gatgaagcgg acgccttcct tcggaagcga gccaccgaga agataagcga
     2701 ggacctcagg gccacactga acgccttcct gtaccgcacg ggccagcaca gcaacaagtt
     2761 catgctggtc ctggccagca accaaccaga gcagttcgac tgggccatca atgaccgcat
     2821 caatgagatg gtccacttcg acctgccagg gcaggaggaa cgggagcgcc tggtgagaat
     2881 gtattttgac aagtatgttc ttaagccggc cacagaagga aagcagcgcc tgaagctggc
     2941 ccagtttgac tacgggagga agtgctcgga ggtcgctcgg ctgacggagg gcatgtcggg
     3001 ccgggagatc gctcagctgg ccgtgtcctg gcaggccacg gcgtatgcct ccgaggacgg
     3061 ggtcctgacc gaggccatga tggacacccg cgtgcaagat gctgtccagc agcaccagca
     3121 gaagatgtgc tggctgaagg cggaagggcc tgggcgtggg gacgagccct ccccatcctg
     3181 agctagcgtt aacacgcgtt gtacatttta cgtatctaga gctcgaggtc gaccaattcc
     3241 ggttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     3301 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     3361 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     3421 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     3481 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     3541 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     3601 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     3661 gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     3721 ttgaaaaaca cgatgataat accatgaaaa agcctgaact caccgcgacg tctgtcgaga
     3781 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
     3841 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
     3901 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
     3961 cgattccgga agtgcttgac attggggaat ttagcgagag cctgacctat tgcatctccc
     4021 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
     4081 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
     4141 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
     4201 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt
     4261 ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc
     4321 acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag
     4381 cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct
     4441 tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc
     4501 atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc
     4561 aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat
     4621 gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa
     4681 gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc
     4741 ccagcactcg tccgagggca aaggaataga gtagatgccg accgggatct atcgatagat
     4801 cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4861 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4921 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     4981 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5041 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5101 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5161 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     5221 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5281 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5341 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc
     5401 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     5461 gggactggaa gggctaattc actcccaacg aagacaagat cacctgcagg acaggcgcgc
     5521 cctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     5581 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     5641 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     5701 agtgtggaaa atctctagca cccgggcgat taaggaaagg gctagatcat tcttgaagac
     5761 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     5821 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5881 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     5941 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6001 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6061 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6121 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6181 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     6241 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6301 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6361 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6421 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6481 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6541 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6601 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6661 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6721 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6781 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     6841 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     6901 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6961 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7021 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7081 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7141 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7201 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7261 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7321 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7381 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7441 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7501 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7561 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7621 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7681 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7741 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     7801 ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag aggccgaggc
     7861 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     7921 ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     7981 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     8041 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     8101 acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8161 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     8221 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
     8281 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
     8341 tatgtactac aaacttagta gttggaaggg ctaattcact cccaaagaag acaagatatc
     8401 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     8461 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     8521 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     8581 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     8641 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
     8701 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
     8761 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
     8821 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     8881 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     8941 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     9001 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     9061 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     9121 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     9181 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     9241 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     9301 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     9361 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     9421 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     9481 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
     9541 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     9601 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     9661 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     9721 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     9781 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     9841 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     9901 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     9961 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
    10021 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
    10081 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
    10141 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
    10201 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
    10261 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
    10321 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
    10381 ggtatcgccg aattcacaaa t
//
