LOCUS       sequence-256953-       10565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10565)
  AUTHORS   Addis RC, Ifkovits JL, Pinto F, Kellam LD, Esteso P, Rentschler S, 
            Christoforou N, Epstein JA, Gearhart JD
  TITLE     Optimization of direct fibroblast reprogramming to cardiomyocytes 
            using calcium activity as a functional measure of success.
  JOURNAL   J Mol Cell Cardiol. 2013 Apr 13;60C:97-106. doi: 
            10.1016/j.yjmcc.2013.04.004.
  PUBMED    23591016
REFERENCE   2  (bases 1 to 10565)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Mol Cell 
            Cardiol."; date: "2013-04-13"; volume: "60C:97-106. doi"; pages: " 
            10.1016/j.yjmcc.2013.04.004"
COMMENT     Sequence Label: TroponinT-GCaMP5-Zeo
FEATURES             Location/Qualifiers
     source          1..10565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     3220..3239
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3220..3238
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3270..3294
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3295..3303,3304..3321,3322..3324,3325..3357,3358..3360,
                     3361..3384,3385..3411,3412..3468,3469..3474,3475..3744,
                     3745..3762,3763..4197,4198..4203,4204..4644)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIG
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(3295..3303,3304..3321,3322..3324,3325..3357,3358..3360,
                     3361..3384,3385..3411,3412..3468,3469..3474,3475..3744,
                     3745..3762,3763..4197,4198..4203,4204..4644)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIG
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      3391..3400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3681..3702
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3808..3829)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4069..4088)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             4654..4707
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    complement(5089..5113)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5139..5180
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5190..5207
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    5257..5845
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5310..5330)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5728..5739)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5848..5864)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5849..5865)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6370..6550
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6576..6593)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6582..6806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6852..7280
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6939..6958)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7149..7170
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7289..7309)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7294..7623
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7474..7609
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7536..7555
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7671..7718
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7737..8111
                     /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    8241..8362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8278..8297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8332..8351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8411..8427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8411..8427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8424..8446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8435..8451
                     /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(8459..8465,8466..8483,8484..8489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8504..8525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8642..8659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8813..9401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8893..8912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9572..10363,10364..10432))
                     /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     10195..10214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10433..10537)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta agtcatggag aagacccacc ttgcagatgt cctcactggg
     2641 gctggcagag ccggcaacct gcccaaggct gctcagtcca ttaggagcca gtagcctgga
     2701 agatgtcttt acccccagca tcagttcaag tggagcagca cataactctt gccctctgcc
     2761 ttccaagatt ctggtgctga gacttatgga gtgtcttgga ggttgccttc tgccccccaa
     2821 ccctgctccc agctggccct cccaggcctg ggttgctggc ctctgcttta tcaggattct
     2881 caagagggac agctggttta tgttgcatga ctgttccctg catatctgct ctggttttaa
     2941 atagcttatc tgagcagctg gaggaccaca tgggcttata tggcgtgggg tacatgttcc
     3001 tgtagccttg tccctggcac ctgccaaaat agcagccaac accccccacc cccaccgcca
     3061 tccccctgcc ccacccgtcc cctgtcgcac attcctccct ccgcagggct ggctcaccag
     3121 gccccagccc acatgcctgc ttaaagccct ctccatcctc tgcctcaccc agtccccgct
     3181 gagactgagc agacgcctcc aggatccgcc accctcgagt aatacgactc actataggga
     3241 gacccaagct ggctaggtaa gcttgatcaa caagtttgta caaaaaagca ggctatgggt
     3301 tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg
     3361 gatctgtacg acgatgacga taaggatctc gccaccatgg tcgactcgtc acgtcgtaag
     3421 tggaataaga caggtcacgc agtcagagct ataggtcggc tgagctcact cgagaacgtc
     3481 tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac
     3541 atcgaggacg gcggcgtgca gctcgcctac cactaccagc agaacacccc catcggcgac
     3601 ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact ttcgaaagac
     3661 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3721 ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag caagggcgag
     3781 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3841 aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct gaccctgaag
     3901 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3961 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     4021 tccgccatgc ccgagggcta catccaggag cgcaccatct tcttcaagga cgacggcaac
     4081 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgggctg
     4141 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacctg
     4201 ccggaccaac tgactgaaga gcagatcgca gaatttaaag aggctttctc cctatttgac
     4261 aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg gtctctgggg
     4321 cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac
     4381 ggcacaatcg acttccctga gttcctgaca atgatggcaa gaaaaatgaa atacacagac
     4441 agtgaagaag aaattagaga agcgttccgt gtgtttgata aggatggcaa tggctacatc
     4501 agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac agatgaagag
     4561 gttgatgaaa tgatcaggga agcagacatc gatggggacg gtcaggtaaa ctacgaagag
     4621 tttgtacaaa tgatgacagc gaagggatcc ggagagggca gaggaagtct tctaacatgc
     4681 ggtgacgtgg aggagaatcc cggcccttcc ggtatggcca aactcacttc tgcagtccca
     4741 gtcctcacag caagggatgt tgcaggggct gtagagttct ggactgacag attaggattc
     4801 tccagagact ttgttgaaga tgattttgct ggtgttgtca gagatgatgt caccctcttc
     4861 atctcagcag ttcaggacca agttgtccct gacaacaccc ttgcttgggt ctgggtcaga
     4921 ggcctagatg agctttatgc agaatggtca gaagtagtca gcacaaattt cagggatgcc
     4981 tctggcccag ccatgacaga aattggtgaa caaccttggg gaagggaatt tgccctcaga
     5041 gaccctgctg gaaattgtgt ccattttgta gctgaggaac aggactagac ccagctttct
     5101 tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc
     5161 tcctcggtct cgattctacg cgtaccggtc atcatcacca tcaccattga gtttaaaccc
     5221 gctgatcagc ctcaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     5281 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5341 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5401 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5461 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5521 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5581 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5641 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     5701 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5761 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5821 atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgagacct agaaaaacat
     5881 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     5941 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     6001 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6061 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     6121 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     6181 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     6241 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     6301 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     6361 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6421 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6481 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6541 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     6601 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     6661 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     6721 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     6781 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     6841 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6901 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6961 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7021 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     7081 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7141 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     7201 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7261 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     7321 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     7381 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7441 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     7501 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     7561 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     7621 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     7681 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     7741 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     7801 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     7861 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     7921 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     7981 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     8041 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     8101 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     8161 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     8221 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     8281 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8341 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     8401 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     8461 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     8521 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     8581 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8641 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8701 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8761 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8821 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8881 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8941 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     9001 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     9061 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     9121 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     9181 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     9241 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9301 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9361 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9421 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9601 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9661 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9721 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     9781 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9841 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9901 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9961 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    10021 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    10081 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    10141 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    10201 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    10261 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10321 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10381 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    10441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    10501 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    10561 ctgac
//
