LOCUS       sequence-451727-        9845 bp    DNA     circular SYN 15-APR-2025
DEFINITION  mT2Del_EPACdDEPCD_Q270E_M312L_L777A_K778A_tdBlackcp173Venus(ST) 
            biosensor to measure realtime changes in FRET reflecting 
            fluctuations in cAMP levels in living cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9845)
  AUTHORS   Zanetti G, Klarenbeek JB, Jalink K
  TITLE     Cytosolic-enhanced dark Epac-based FRET sensors allow for 
            intracellular cAMP detection in live cells via FLIM.
  JOURNAL   FEBS Lett. 2024 Dec 31. doi: 10.1002/1873-3468.15093.
  PUBMED    39737677
REFERENCE   2  (bases 1 to 9845)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "FEBS Lett. 
            2024 Dec 31. doi: 10.1002/1873-3468.15093."
COMMENT     Sequence Label: Epac-S-H327
FEATURES             Location/Qualifiers
     source          1..9845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      10..19
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      14..23
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(28..30,31..33,34..546)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIE"
     CDS             2932..3666
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     CDS             3724..4458
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     promoter        complement(4510..4528)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4511..4528)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4548..4565)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4554..4778
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4824..5252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4911..4930)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5121..5142
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5261..5281)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5266..5595
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5446..5581
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5508..5527
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5662..6456
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5716..5735)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6326..6345
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6630..6751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6667..6686)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6721..6740
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6800..6816)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6800..6816)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6813..6835)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6824..6840
                     /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(6848..6854,6855..6872,6873..6878))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6893..6914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7031..7048)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7202..7790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7282..7301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7961..8752,8753..8821))
                     /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     8584..8603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8822..8926)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9001..9020)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9192..9571
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9572..9775
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9726..9746
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9820..9839
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9820..9838
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagcttgccg ccaccatggc ggccgcaatg gtgagcaagg gcgaggagct gttcaccggg
       61 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
      121 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
      181 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc
      241 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
      301 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
      361 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
      421 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actactttag cgacaacgtc
      481 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
      541 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
      601 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
      661 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc ccgatatcag
      721 cccgtgggaa ctcatgagat ggaggaggag ttggccgaag ctgtggccct gctctcccag
      781 cgggggcctg acgccctgct cactgtggca cttcgaaagc ccccaggtca gcgcacggat
      841 gaagagctgg acctcatctt tgaggagctg ctgcacatca aggctgtggc ccacctctcc
      901 aactcggtga agcgagaatt agcggctgtt ctgctctttg aaccacacag caaggcaggg
      961 accgtgttgt tcagccaggg ggacaagggc acttcgtggt acattatctg gaagggatct
     1021 gtcaacgtgg tgacccatgg caaggggctg gtcaccaccc tgcatgaggg agatgatttt
     1081 ggagagctgg ctctggtgaa tgatgcaccc cgggcagcca ccatcatcct gcgagaagac
     1141 aactgtcatt tcctgcgtgt ggacaagcag gacttcaacc gtatcatcaa ggatgtggag
     1201 gcaaagacct tgcggctgga agaacatggc aaagtggtgc tggtgctgga gagagcctct
     1261 cagggcgccg gcccttcccg acccccaacc ccaggcagga accggtatac agtgatgtct
     1321 ggcaccccag agaagatcct agagcttctg ttggaggcca tgggaccaga ttccagtgct
     1381 catgacccaa cagagacatt cctcagcgac ttcctcctga cccacagggt cttcatgccc
     1441 agcgcccaac tctgcgctgc ccttctgcac cacttccatg tggagcctgc gggtggcagc
     1501 gagcaggagc gcagcaccta cgtctgcaac aagaggcagc agatcttgcg gctggtcagc
     1561 cagtgggtgg ccctgtatgg ctccatgctc cacactgacc ctgtggccac cagcttcctc
     1621 cagaaactct cagacctggt gggcagggac acccgactca gcaacctgct gagggagcag
     1681 tggccagaga ggcggcgatg ccacaggttg gagaatggct gtgggaatgc atctcctcag
     1741 atgaaggccc ggaacttgcc tgtttggctc cccaaccagg acgagcccct tcctggcagc
     1801 agctgtgcca tccaagttgg ggataaagtc ccctatgaca tctgccggcc agaccactca
     1861 gtgttgaccc tgcagctgcc tgtgacagcc tccgtgagag aggtgatggc agcgttggcc
     1921 caggaggatg gctggaccaa ggggcaggtg ctggtgaagg tcaattctgc aggtgatgcc
     1981 attggcctgc agccagatgc ccgtggtgtg gccacatctc tggggctcaa tgagcgtctc
     2041 tttgttgtca acccacagga agcgcatgag ctgatcccac accctgacca gctggggccc
     2101 actgtgggct ctgctgaggg gctggacctg gtgagtgcca aggacctggc aggccagctg
     2161 acggaccacg actggagcct cttcaacagt atccaccagg tggagctgat ccactatgtg
     2221 ctgggccccc agcatctgcg ggatgtcacc accgccaacc tggagcgctt catgcgccgc
     2281 ttcaatgagc tgcagtactg ggtggccacc gagctgtgtc tctgccccgt gcccggcccc
     2341 cgggcccagc tgctcaggaa gttcattaag ctggcggccc acctcaagga gcagaagaat
     2401 ctcaattcct tctttgccgt catgtttggc ctcagcaact cggccatcag ccgcctagcc
     2461 cacacctggg agcggctgcc ccacaaagtc cggaagctgt actccgccct cgagaggctg
     2521 ctggatccct catggaacca ccgggtatac cgactggccc tcgccaagct ctcccctcct
     2581 gtcatcccct tcatgcccct tcttgccgca gacatggccg ccattcatga gggaaaccac
     2641 acactagtgg agaatctcat caactttgag aagatgagaa tgatggccag agccgcgcgg
     2701 atgctgcacc actgccgaag ccacaaccct gtgcctctct caccactcag aagccgagtt
     2761 tcccacctcc acgaggacag ccaggtggcg aggatttcca catgctcgga gcagtccctg
     2821 agcacccgga gtccagccag cacctgggct tatgtccagc agctgaaggt cattgacaac
     2881 cagcgggaac tctcccgcct ctcccgagag ctggagccag ctagcgagct catggacggc
     2941 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3001 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
     3061 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3121 gagctgtaca agggtggcag cggtggcatg gtgagcaagg gcgaggagct gttcaccggg
     3181 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     3241 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat ctgcaccacc
     3301 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg cctgcagtgc
     3361 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     3421 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     3481 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     3541 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actggaacag ccacaacgtc
     3601 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     3661 atcgagccga attttgtctt cctgatcggc gccgcaggaa tactcttcgt atctagcgag
     3721 ctcatggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     3781 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     3841 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3901 ctcggcatgg acgagctgta caagggtggc agcggtggca tggtgagcaa gggcgaggag
     3961 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     4021 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     4081 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgggctac
     4141 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     4201 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     4261 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4321 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactggaac
     4381 agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc caacttcaag
     4441 atccgccaca acatcgagcc gaattccctc gaggttaacg cggcgcgcct ctaatctaga
     4501 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     4561 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4621 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4681 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4741 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4801 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4861 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4921 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4981 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5041 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5101 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5161 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5221 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     5281 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5341 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5401 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5461 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5521 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5581 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     5641 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5701 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5761 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5821 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5881 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5941 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     6001 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     6061 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     6121 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6181 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6241 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6301 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6361 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6421 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     6481 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     6541 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     6601 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6661 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6721 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6781 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6841 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6901 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6961 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7021 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7081 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7141 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7621 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8941 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     9001 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     9061 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     9121 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     9181 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     9241 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     9301 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     9361 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     9421 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     9481 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     9541 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     9601 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     9661 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     9721 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     9781 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     9841 gaccc
//
