LOCUS       sequence-451793-        9053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mT2Del_EPACdDEPCD_Q270E_M312L_L777A_K778A_tdcp173Venus(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 9053)
  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 9053)
  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: "FEBS Lett. 
            2024 Dec 31. doi: 10.1002/1873-3468.15093."
COMMENT     Sequence Label: Epac-S-H323
FEATURES             Location/Qualifiers
     source          1..9053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      450..1038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     939..958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1192..1209
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1326..1347
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1362..1367,1368..1385,1386..1392)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1400..1416
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1405..1427
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1424..1440
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1424..1440
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1489..1610
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1500..1519)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1554..1573
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1784..2578)
                     /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(1895..1914)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2505..2524
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2645..2974)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2659..2794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2713..2732)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2959..2979
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2988..3416)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3098..3119)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3310..3329
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3462..3686
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3675..3692
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3712..3730
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3712..3729
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3782..4516)
                     /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
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     CDS             complement(join(6902..7414,7415..7417,7418..7420))
                     /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"
     regulatory      7425..7434
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      7429..7438
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(7454..7473)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7455..7473)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(7518..7721)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7547..7567)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7722..8101
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     8273..8292
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        8367..8471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8472..8540,8541..9053,1..279)
                     /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     complement(8690..8709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
       61 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
      121 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
      181 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
      241 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
      301 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
      361 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
      421 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
      481 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      541 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
      601 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      661 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      721 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      781 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      841 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      901 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      961 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1021 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1081 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1141 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1201 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1261 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     1321 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     1381 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     1441 catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata agatacattg
     1501 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1561 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg
     1621 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     1681 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     1741 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     1801 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     1861 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     1921 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     1981 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     2041 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     2101 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     2161 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     2221 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     2281 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     2341 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     2401 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     2461 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     2521 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     2581 gaaacgatcc tcatcctgtc tcttgatcag atccgaaaat ggatatacaa gctcccggga
     2641 gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
     2701 aggcagaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     2761 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg
     2821 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     2881 ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     2941 agcctgggga ctttccacac cctaactgac acacattcca cagaattaat tcgcgttaaa
     3001 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3061 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3121 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3181 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3241 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3301 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3361 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg
     3421 atacccccta gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc
     3481 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
     3541 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
     3601 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
     3661 agatggctgg caactagaag gcacagtcga ggctgatcag cgagctctag catttaggtg
     3721 acactataga atagggccct ctagattaga ggcgcgccgc gttaacctcg agggaattcg
     3781 gctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     3841 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     3901 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     3961 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     4021 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     4081 gggcgaagca ctgcaggccg tagcccaggg tggtcacgag ggtgggccag ggcacgggca
     4141 gcttgccggt ggtgcagatc agcttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     4201 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     4261 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat gccaccgctg ccacccttgt
     4321 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
     4381 gatcgcgctt ctcgttgggg tctttgctca gggcggactg gtagctcagg tagtggttgt
     4441 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
     4501 gcacgccgcc gtccatgagc tcgctagctg gctccagctc tcgggagagg cgggagagtt
     4561 cccgctggtt gtcaatgacc ttcagctgct ggacataagc ccaggtgctg gctggactcc
     4621 gggtgctcag ggactgctcc gagcatgtgg aaatcctcgc cacctggctg tcctcgtgga
     4681 ggtgggaaac tcggcttctg agtggtgaga gaggcacagg gttgtggctt cggcagtggt
     4741 gcagcatccg cgcggctctg gccatcattc tcatcttctc aaagttgatg agattctcca
     4801 ctagtgtgtg gtttccctca tgaatggcgg ccatgtctgc ggcaagaagg ggcatgaagg
     4861 ggatgacagg aggggagagc ttggcgaggg ccagtcggta tacccggtgg ttccatgagg
     4921 gatccagcag cctctcgagg gcggagtaca gcttccggac tttgtggggc agccgctccc
     4981 aggtgtgggc taggcggctg atggccgagt tgctgaggcc aaacatgacg gcaaagaagg
     5041 aattgagatt cttctgctcc ttgaggtggg ccgccagctt aatgaacttc ctgagcagct
     5101 gggcccgggg gccgggcacg gggcagagac acagctcggt ggccacccag tactgcagct
     5161 cattgaagcg gcgcatgaag cgctccaggt tggcggtggt gacatcccgc agatgctggg
     5221 ggcccagcac atagtggatc agctccacct ggtggatact gttgaagagg ctccagtcgt
     5281 ggtccgtcag ctggcctgcc aggtccttgg cactcaccag gtccagcccc tcagcagagc
     5341 ccacagtggg ccccagctgg tcagggtgtg ggatcagctc atgcgcttcc tgtgggttga
     5401 caacaaagag acgctcattg agccccagag atgtggccac accacgggca tctggctgca
     5461 ggccaatggc atcacctgca gaattgacct tcaccagcac ctgccccttg gtccagccat
     5521 cctcctgggc caacgctgcc atcacctctc tcacggaggc tgtcacaggc agctgcaggg
     5581 tcaacactga gtggtctggc cggcagatgt cataggggac tttatcccca acttggatgg
     5641 cacagctgct gccaggaagg ggctcgtcct ggttggggag ccaaacaggc aagttccggg
     5701 ccttcatctg aggagatgca ttcccacagc cattctccaa cctgtggcat cgccgcctct
     5761 ctggccactg ctccctcagc aggttgctga gtcgggtgtc cctgcccacc aggtctgaga
     5821 gtttctggag gaagctggtg gccacagggt cagtgtggag catggagcca tacagggcca
     5881 cccactggct gaccagccgc aagatctgct gcctcttgtt gcagacgtag gtgctgcgct
     5941 cctgctcgct gccacccgca ggctccacat ggaagtggtg cagaagggca gcgcagagtt
     6001 gggcgctggg catgaagacc ctgtgggtca ggaggaagtc gctgaggaat gtctctgttg
     6061 ggtcatgagc actggaatct ggtcccatgg cctccaacag aagctctagg atcttctctg
     6121 gggtgccaga catcactgta taccggttcc tgcctggggt tgggggtcgg gaagggccgg
     6181 cgccctgaga ggctctctcc agcaccagca ccactttgcc atgttcttcc agccgcaagg
     6241 tctttgcctc cacatccttg atgatacggt tgaagtcctg cttgtccaca cgcaggaaat
     6301 gacagttgtc ttctcgcagg atgatggtgg ctgcccgggg tgcatcattc accagagcca
     6361 gctctccaaa atcatctccc tcatgcaggg tggtgaccag ccccttgcca tgggtcacca
     6421 cgttgacaga tcccttccag ataatgtacc acgaagtgcc cttgtccccc tggctgaaca
     6481 acacggtccc tgccttgctg tgtggttcaa agagcagaac agccgctaat tctcgcttca
     6541 ccgagttgga gaggtgggcc acagccttga tgtgcagcag ctcctcaaag atgaggtcca
     6601 gctcttcatc cgtgcgctga cctgggggct ttcgaagtgc cacagtgagc agggcgtcag
     6661 gcccccgctg ggagagcagg gccacagctt cggccaactc ctcctccatc tcatgagttc
     6721 ccacgggctg atatcgggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     6781 cgttggggtc tttgctcagc ttggactggg tgctcaggta gtggttgtcg ggcagcagca
     6841 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgccgccgt
     6901 cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc ttgtcggcgg
     6961 tgatatagac gttgtcgcta aagtagttgt actccagctt gtgccccagg atgttgccgt
     7021 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     7081 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     7141 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     7201 gggcgaagca ctgcacgccc caggacaggg tggtcacgag ggtgggccag ggcacgggca
     7261 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     7321 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     7381 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat tgcggccgcc atggtggcgg
     7441 caagcttggg tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt
     7501 tctctagtta gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc
     7561 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     7621 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     7681 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     7741 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     7801 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     7861 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     7921 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     7981 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     8041 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     8101 caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga
     8161 ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac
     8221 tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca
     8281 gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt
     8341 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     8401 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     8461 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     8521 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     8581 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     8641 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     8701 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     8761 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     8821 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     8881 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     8941 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     9001 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggc
//
