Ef1a_Large T-antigen_Ires_Puro

LOCUS       Ef1a_Large T-antigen_Ires_Puro               11672 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Ef1a_Large T-antigen_Ires_Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11672)
  AUTHORS   Mali P, Ye Z, Hommond HH, Yu X, Lin J, Chen G, Zou J, Cheng L
  TITLE     Improved efficiency and pace of generating induced pluripotent stem 
            cells from human adult and fetal fibroblasts.
  JOURNAL   Stem Cells. 2008 Aug . 26(8):1998-2005.
  PUBMED    18511599
REFERENCE   2  (bases 1 to 11672)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11672
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..2127
                     /codon_start=1
                     /product="large T antigen from SV40 (simian virus 40)"
                     /label=large T antigen
                     /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK
                     GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEIPTYGTDEWEQWWNAFNEENLF
                     CSEEMPSSDDEATADSQHSTPPKKKRKVEDPKDFPSELLSFLSHAVFSNRTLACFAIYT
                     TKEKAALLYKKIMEKYSVTFISRHNSYNHNILFFLTPHRHRVSAINNYAQKLCTFSFLI
                     CKGVNKEYLMYSALTRDPFSVIEESLPGGLKEHDFNPEEAEETKQVSWKLVTEYAMETK
                     CDDVLLLLGMYLEFQYSFEMCLKCIKKEQPSHYKYHEKHYANAAIFADSKNQKTICQQA
                     VDTVLAKKRVDSLQLTREQMLTNRFNDLLDRMDIMFGSTGSADIEEWMAGVAWLHCLLP
                     KMDSVVYDFLKCMVYNIPKKRYWLFKGPIDSGKTTLAAALLELCGGKALNVNLPLDRLN
                     FELGVAIDQFLVVFEDVKGTGGESRDLPSGQGINNLDNLRDYLDGSVKVNLEKKHLNKR
                     TQIFPPGIVTMNEYSVPKTLQARFVKQIDFRPKDYLKHCLERSEFLLEKRIIQSGIALL
                     LMLIWYRPVAEFAQSIQSRIVEWKERLDKEFSLSVYQKMKFNVAMGIGVLDWLRNSDDD
                     DEDSQENADKNEDGGEKNMEDSGHETGIDSQSQGSFQAPQSSQSVHDHNQPYHICRGFT
                     CFKKPPTPPPEPET"
     misc_feature    2159..2745
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2326..2343)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2553..2572
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2746..3345
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2746..2765)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3242..3262
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    3459..3492
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     LTR             3517..3697
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3723..3740)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3729..3953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3999..4427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4086..4105)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4296..4317
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4436..4456)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4441..4770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4621..4756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4683..4702
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4818..4865
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4884..5258
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5388..5509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5425..5444)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5479..5498
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5558..5574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5558..5574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5571..5593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5582..5598
                     /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)."
     promoter        complement(5606..5636)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5651..5672
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5789..5806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5960..6548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6040..6059)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6719..7579)
                     /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     7342..7361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7580..7684)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7759..7778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7950..8329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8331..8529
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8484..8504
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             8547..8727
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8774..8899
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9392..9625
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9810..9854
                     /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    10152..10269
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        10405..11583
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          10636..11574
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     11531..11551
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     11600..11619
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        11600..11618
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
       61 aggagtgcct gggggaatat tcctctgatg agaaaggcat atttaaaaaa atgcaaggag
      121 tttcatcctg ataaaggagg agatgaagaa aaaatgaaga aaatgaatac tctgtacaag
      181 aaaatggaag atggagtaaa atatgctcat caacctgact ttggaggctt ctgggatgca
      241 actgagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc ctttaatgag
      301 gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac tgctgactct
      361 caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga ctttccttca
      421 gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc ttgctttgct
      481 atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga aaaatattct
      541 gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt tcttactcca
      601 cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac ctttagcttt
      661 ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac tagagatcca
      721 ttttctgtta ttgaggaaag tttgccaggt gggttaaagg agcatgattt taatccagaa
      781 gaagcagagg aaactaaaca agtgtcctgg aagcttgtaa cagagtatgc aatggaaaca
      841 aaatgtgatg atgtgttgtt attgcttggg atgtacttgg aatttcagta cagttttgaa
      901 atgtgtttaa aatgtattaa aaaagaacag cccagccact ataagtacca tgaaaagcat
      961 tatgcaaatg ctgctatatt tgctgacagc aaaaaccaaa aaaccatatg ccaacaggct
     1021 gttgatactg ttttagctaa aaagcgggtt gatagcctac aattaactag agaacaaatg
     1081 ttaacaaaca gatttaatga tcttttggat aggatggata taatgtttgg ttctacaggc
     1141 tctgctgaca tagaagaatg gatggctgga gttgcttggc tacactgttt gttgcccaaa
     1201 atggattcag tggtgtatga ctttttaaaa tgcatggtgt acaacattcc taaaaaaaga
     1261 tactggctgt ttaaaggacc aattgatagt ggtaaaacta cattagcagc tgctttgctt
     1321 gaattatgtg gggggaaagc tttaaatgtt aatttgccct tggacaggct gaactttgag
     1381 ctaggagtag ctattgacca gtttttagta gtttttgagg atgtaaaggg cactggaggg
     1441 gagtccagag atttgccttc aggtcaggga attaataacc tggacaattt aagggattat
     1501 ttggatggca gtgttaaggt aaacttagaa aagaaacacc taaataaaag aactcaaata
     1561 tttccccctg gaatagtcac catgaatgag tacagtgtgc ctaaaacact gcaggccaga
     1621 tttgtaaaac aaatagattt taggcccaaa gattatttaa agcattgcct ggaacgcagt
     1681 gagtttttgt tagaaaagag aataattcaa agtggcattg ctttgcttct tatgttaatt
     1741 tggtacagac ctgtggctga gtttgctcaa agtattcaga gcagaattgt ggagtggaaa
     1801 gagagattgg acaaagagtt tagtttgtca gtgtatcaaa aaatgaagtt taatgtggct
     1861 atgggaattg gagttttaga ttggctaaga aacagtgatg atgatgatga agacagccag
     1921 gaaaatgctg ataaaaatga agatggtggg gagaagaaca tggaagactc agggcatgaa
     1981 acaggcattg attcacagtc ccaaggctca tttcaggccc ctcagtcctc acagtctgtt
     2041 catgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2101 acacctcccc ctgaacctga aacataagtt aaccgataag ggcgaattcc gccccccccc
     2161 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     2221 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     2281 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     2341 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     2401 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     2461 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     2521 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     2581 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     2641 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     2701 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatgac cgagtacaag
     2761 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     2821 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     2881 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     2941 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     3001 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     3061 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3121 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     3181 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     3241 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     3301 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
     3361 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cgaacaaaag aaaaggggac
     3421 tggaagggct aattcactcc caacgaagac aagatatcat aacttcgtat agcatacatt
     3481 atacgaagtt atcggctagc tggtccggac tgtactgggt ctctctggtt agaccagatc
     3541 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     3601 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     3661 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
     3721 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3781 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3841 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3901 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3961 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     4021 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4081 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4141 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4201 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4261 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4321 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4381 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4441 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4501 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4561 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4621 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4681 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4741 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4801 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
     4861 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
     4921 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
     4981 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
     5041 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
     5101 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
     5161 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
     5221 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
     5281 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5341 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     5401 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5461 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     5521 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5581 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5641 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5701 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5761 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5821 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5881 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5941 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6001 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6061 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6121 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6181 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6241 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6301 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6361 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6421 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6481 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6721 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6781 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6841 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6901 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6961 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7021 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7081 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7141 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7201 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7261 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7321 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7381 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7441 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7501 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7561 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7621 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7681 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7741 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     7801 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     7861 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     7921 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     7981 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     8041 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     8101 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     8161 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     8221 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     8281 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     8341 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     8401 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     8461 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     8521 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     8581 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     8641 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     8701 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     8761 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     8821 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     8881 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     8941 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     9001 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     9061 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     9121 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     9181 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     9241 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     9301 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     9361 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     9421 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     9481 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     9541 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     9601 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     9661 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     9721 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     9781 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     9841 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     9901 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     9961 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
    10021 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
    10081 ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta
    10141 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
    10201 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
    10261 caaaattttc gggtttatta cagggacagc agagatccag tttggttaat taatcgataa
    10321 gctttgcaaa gatggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc
    10381 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
    10441 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
    10501 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
    10561 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
    10621 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
    10681 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
    10741 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
    10801 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
    10861 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
    10921 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
    10981 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
    11041 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
    11101 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
    11161 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
    11221 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
    11281 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
    11341 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
    11401 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
    11461 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
    11521 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
    11581 tgaggaatta gcttggtact aatacgactc actataggga gacccaagct ggctaggtaa
    11641 gcttgatatc gaattcgccc ttatcgtcta ga
//