LOCUS       sequence-347242-       10051 bp    DNA     circular SYN 14-APR-2025
DEFINITION  doxycycline-inducible overexpression of AKT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10051)
  AUTHORS   Romero-Tejeda M, Fonoudi H, Weddle CJ, DeKeyser JM, Lenny B, 
            Fetterman KA, Magdy T, Sapkota Y, Epting C, Burridge PW
  TITLE     A novel transcription factor combination for direct reprogramming to
            a spontaneously contracting human cardiomyocyte-like state.
  JOURNAL   J Mol Cell Cardiol. 2023 Jul 6:S0022-2828(23)00106-2. doi: 
            10.1016/j.yjmcc.2023.06.005.
  PUBMED    37421991
REFERENCE   2  (bases 1 to 10051)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Mol Cell 
            Cardiol. 2023 Jul 6:S0022-2828(23)00106-2. doi: 
            10.1016/j.yjmcc.2023.06.005."
COMMENT     Sequence Label: tetO-AKT1
FEATURES             Location/Qualifiers
     source          1..10051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..68)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        240..619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             837..1017
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1064..1189
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1682..1915
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2100..2144
                     /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    2442..2559
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2612..2630,2631..2653,2654..2672,2673..2695,2696..2714,
                     2715..2737,2738..2756,2757..2779,2780..2798,2799..2821,
                     2822..2840,2841..2863,2864..2882)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2915..2953
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2950..2974
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3053..3072
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3053..3071
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3103..3127
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3133..4572
                     /codon_start=1
                     /gene="AKT1"
                     /product="human serine/threonine protein kinase activated 
                     by growth factors"
                     /label=Akt1
                     /translation="MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQ
                     TVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDA
                     KEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGTA"
     protein_bind    complement(4577..4601)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4627..4668
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4678..4695
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4745..5333
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4798..4818)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5216..5227)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5336..5352)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5337..5353)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5858..6038
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6064..6081)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6070..6294
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6340..6768
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6427..6446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6637..6658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6777..6797)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6782..7111
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6962..7097
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7024..7043
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7159..7206
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7225..7599
                     /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    7729..7850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7766..7785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7820..7839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7899..7915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7899..7915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7912..7934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7923..7939
                     /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(join(7947..7953,7954..7971,7972..7977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7992..8013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8130..8147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8301..8889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8381..8400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9060..9851,9852..9920))
                     /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     9683..9702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9921..10025)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
       61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
      121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
      181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      781 gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt
      841 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
      901 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
      961 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     1021 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     1081 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     1141 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     1201 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     1261 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     1321 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     1381 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     1441 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     1501 caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata
     1561 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     1621 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     1681 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     1741 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     1801 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     1861 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     1921 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     1981 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     2041 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     2101 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     2161 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     2221 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     2281 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     2341 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc
     2401 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     2461 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     2521 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     2581 agagatccag tttggttatc gagtttacca ctccctatca gtgatagaga aaagtgaaag
     2641 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct
     2701 atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag
     2761 tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca
     2821 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
     2881 gaaaagtgaa agtcgagctc ggtacccggg tcgaggtagg cgtgtacggt gggaggccta
     2941 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     3001 tgacctccat agaagacacc gggaccgatc cagcctccgc ggccccgaat tgtaatacga
     3061 ctcactatag ggagacccaa gctggctagg taagcttgat caacaagttt gtacaaaaaa
     3121 gcaggcttca ccatgagcga cgtggctatt gtgaaggagg gttggctgca caaacgaggg
     3181 gagtacatca agacctggcg gccacgctac ttcctcctca agaatgatgg caccttcatt
     3241 ggctacaagg agcggccgca ggatgtggac caacgtgagg ctcccctcaa caacttctct
     3301 gtggcgcagt gccagctgat gaagacggag cggccccggc ccaacacctt catcatccgc
     3361 tgcctgcagt ggaccactgt catcgaacgc accttccatg tggagactcc tgaggagcgg
     3421 gaggagtgga caaccgccat ccagactgtg gctgacggcc tcaagaagca ggaggaggag
     3481 gagatggact tccggtcggg ctcacccagt gacaactcag gggctgaaga gatggaggtg
     3541 tccctggcca agcccaagca ccgcgtgacc atgaacgagt ttgagtacct gaagctgctg
     3601 ggcaagggca ctttcggcaa ggtgatcctg gtgaaggaga aggccacagg ccgctactac
     3661 gccatgaaga tcctcaagaa ggaagtcatc gtggccaagg acgaggtggc ccacacactc
     3721 accgagaacc gcgtcctgca gaactccagg caccccttcc tcacagccct gaagtactct
     3781 ttccagaccc acgaccgcct ctgctttgtc atggagtacg ccaacggggg cgagctgttc
     3841 ttccacctgt cccgggagcg tgtgttctcc gaggaccggg cccgcttcta tggcgctgag
     3901 attgtgtcag ccctggacta cctgcactcg gagaagaacg tggtgtaccg ggacctcaag
     3961 ctggagaacc tcatgctgga caaggacggg cacattaaga tcacagactt cgggctgtgc
     4021 aaggagggga tcaaggacgg tgccaccatg aagacctttt gcggcacacc tgagtacctg
     4081 gcccccgagg tgctggagga caatgactac ggccgtgcag tggactggtg ggggctgggc
     4141 gtggtcatgt acgagatgat gtgcggtcgc ctgcccttct acaaccagga ccatgagaag
     4201 ctttttgagc tcatcctcat ggaggagatc cgcttcccgc gcacgcttgg tcccgaggcc
     4261 aagtccttgc tttcagggct gctcaagaag gaccccaagc agaggcttgg cgggggctcc
     4321 gaggacgcca aggagatcat gcagcatcgc ttctttgccg gtatcgtgtg gcagcacgtg
     4381 tacgagaaga agctcagccc acccttcaag ccccaggtca cgtcggagac tgacaccagg
     4441 tattttgatg aggagttcac ggcccagatg atcaccatca caccacctga ccaagatgac
     4501 agcatggagt gtgtggacag cgagcgcagg ccccacttcc cccagttctc ctactcggcc
     4561 agcggcacgg cctgaaaccc agctttcttg tacaaagtgg ttgatctaga gggcccgcgg
     4621 ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg taccggtcat
     4681 catcaccatc accattgagt ttaaacccgc tgatcagcct caattcgata tcaagcttat
     4741 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4801 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4861 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4921 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4981 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     5041 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     5101 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     5161 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     5221 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     5281 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     5341 ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     5401 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
     5461 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     5521 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     5581 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc
     5641 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga
     5701 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca
     5761 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt
     5821 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat
     5881 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     5941 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6001 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
     6061 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     6121 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     6181 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     6241 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     6301 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     6361 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6421 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6481 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6541 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6601 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6661 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6721 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     6781 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6841 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     6901 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6961 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     7021 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     7081 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     7141 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     7201 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     7261 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     7321 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     7381 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     7441 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     7501 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     7561 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     7621 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     7681 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     7741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     7801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     7861 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     7921 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     7981 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     8041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     8101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     8161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     8221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     8281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     8341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     8401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     8461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     8521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     8581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     8641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     8701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     8761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     8821 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     8881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     8941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     9001 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     9061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     9121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     9181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     9241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     9301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     9361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     9421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     9481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     9541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     9601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     9661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     9721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     9781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     9841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     9901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     9961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    10021 ccgcgcacat ttccccgaaa agtgccacct g
//
