pInducer20-Blast

LOCUS       pInducer20-Blast       12215 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pInducer20 with added blasticidin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12215)
  AUTHORS   Matson JP, Dumitru R, Coryell P, Baxley RM, Chen W, Twaroski K, 
            Webber BR, Tolar J, Bielinsky AK, Purvis JE, Cook JG
  TITLE     Rapid DNA replication origin licensing protects stem cell 
            pluripotency.
  JOURNAL   Elife. 2017 Nov 17;6. doi: 10.7554/eLife.30473.
  PUBMED    29148972
REFERENCE   2  (bases 1 to 12215)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2017-11-17"; volume: "6. doi"; pages: " 10.7554/eLife.30473"
COMMENT     Sequence Label: pInducer20-Blast
FEATURES             Location/Qualifiers
     source          1..12215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2230..2248,2249..2271,2272..2290,2291..2313,2314..2332,
                     2333..2355,2356..2374,2375..2397,2398..2416,2417..2439,
                     2440..2458,2459..2481,2482..2500)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2533..2571
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2568..2592
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2701..2825
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2850..2855,2856..2873,2874..2880)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2934..3593
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3000..3019)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3935..4240
                     /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     4119..4138
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4281..4405)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4458..4484
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        4570..5781
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5618..5637
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5840..6457,6458..6463,6464..6499,6500..6502,6503..6538,
                     6539..6541,6542..6577,6578..6586)
                     /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"
     misc_feature    6639..7212
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6806..6823)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7033..7052
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7222..7620
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             7634..8428
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             7634..8428
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7688..7707)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8298..8317
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    8605..9193
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8658..8678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9076..9087)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9268..9501
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9620..9638)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9706..9810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9811..9879,9880..10671)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10029..10048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10842..11430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11331..11350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11584..11601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11841..11862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11877..11882,11883..11900,11901..11907)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11915..11931
                     /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     11920..11942
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11939..11955
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11939..11955
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12005..12024
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactaggatc
     2221 ctttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     2281 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     2341 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     2401 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     2461 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
     2521 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
     2581 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     2641 gaccgatcca gcctccgcgg ccccgaacta gtccagtgtg gtggaattct gcagatatca
     2701 acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg atataaatat caatatatta
     2761 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca
     2821 ctatggcggc cgcattaggc accccaggct ttacacttta tgcttccggc tcgtataatg
     2881 tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct aaaatggaga
     2941 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     3001 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     3061 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     3121 ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac ggtgagctgg
     3181 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     3241 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     3301 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     3361 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     3421 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     3481 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     3541 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taaagatctg
     3601 gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta
     3661 taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg ctatgaagca
     3721 gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc
     3781 aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa
     3841 cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac
     3901 ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt acacctataa
     3961 aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg
     4021 gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga
     4081 actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc
     4141 cagtgtgccg gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga
     4201 catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct cccttataca
     4261 cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag tattatgtag
     4321 tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt tacgtttctc
     4381 gttcagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc tcgagtctag
     4441 agggcccgcg gttcgaatac ccatacgacg tcccagacta cgcttagtaa tgattaatta
     4501 aactagaaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agtaacccac
     4561 ccaagatctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg
     4621 cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc
     4681 tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat cagcagaagg
     4741 acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca gagagcggaa
     4801 caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg tggggcggtg
     4861 aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg
     4921 gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg agtagcgggc
     4981 tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg
     5041 agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg
     5101 gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg cttgtgaggc
     5161 gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct
     5221 tgaggccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg ggcaccatct
     5281 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     5341 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     5401 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     5461 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     5521 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     5581 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     5641 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     5701 tttgggtcaa tatgtaattt tcagtgttag actagtaaat tgtccgctaa attctggccg
     5761 tttttggctt ttttgttaga cgaagcttgg taccgagctc ggatctccac cccgtaccgg
     5821 tcctgcagtc gaattcacca tgtctagact ggacaagagc aaagtcataa acggagctct
     5881 ggaattactc aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct
     5941 gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc
     6001 cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc
     6061 atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg
     6121 cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga
     6181 aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct
     6241 gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc
     6301 aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat
     6361 tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat
     6421 atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga
     6481 ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc
     6541 tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcc
     6601 gcggccgcac tagaggaatt ccgcccctct cccccccccc cntctccctc ccccccccct
     6661 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     6721 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     6781 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     6841 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     6901 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     6961 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     7021 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     7081 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     7141 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa
     7201 acacgatgat aagcttaaac catggccaag cctttgtctc aagaagaatc caccctcatt
     7261 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
     7321 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     7381 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     7441 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     7501 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     7561 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     7621 tagccggtcc accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     7681 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     7741 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     7801 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     7861 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     7921 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     7981 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     8041 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     8101 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     8161 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     8221 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     8281 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     8341 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     8401 ctatcgcctt cttgacgagt tcttctgatg tacaagtaaa gcggccgcga ctctagatca
     8461 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     8521 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt ttagtccctc ccaattcgat
     8581 atcaagctta tcgcgataga tcctaatcaa cctctggatt acaaaatttg tgaaagattg
     8641 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     8701 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     8761 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     8821 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     8881 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     8941 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     9001 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     9061 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     9121 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     9181 gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct gtagatctta
     9241 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     9301 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     9361 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     9421 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     9481 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     9541 ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa agggctagat
     9601 cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     9661 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     9721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     9781 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     9841 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     9901 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     9961 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
    10021 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
    10081 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
    10141 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
    10201 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
    10261 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
    10321 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
    10381 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
    10441 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
    10501 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
    10561 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
    10621 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
    10681 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
    10741 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
    10801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
    10861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
    10921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
    10981 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
    11041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
    11101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
    11161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
    11221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
    11281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
    11341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
    11401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
    11461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
    11521 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
    11581 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
    11641 gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg
    11701 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
    11761 gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg
    11821 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    11881 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    11941 ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc actgcattct
    12001 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
    12061 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
    12121 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
    12181 tgtatttgtt aaatatgtac tacaaactta gtagt
//