LOCUS       sequence-356422-       10456 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of the cAMP sensor cAMPFIRE-L under a CAG 
            promotor w/ WPRE translation enhancer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10456)
  AUTHORS   Massengill CI, Bayless-Edwards L, Ceballos CC, Cebul ER, Cahill J, 
            Bharadwaj A, Wilson E, Qin M, Whorton MR, Baconguis I, Ye B, Mao T, 
            Zhong H
  TITLE     Sensitive genetically encoded sensors for population and subcellular
            imaging of cAMP in vivo.
  JOURNAL   Nat Methods. 2022 Nov;19(11):1461-1471. doi: 
            10.1038/s41592-022-01646-5. Epub 2022 Oct 27.
  PUBMED    36303019
REFERENCE   2  (bases 1 to 10456)
  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"; doi: 
            "10.1038/s41592-022-01646-5"; journalName: "Nat Methods"; date: 
            "2022-11"; volume: "19"; issue: "11"; pages: "1461-1471"
COMMENT     Sequence Label: pCAGGS_cAMPFIRE-L
FEATURES             Location/Qualifiers
     source          1..10456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1755..1764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1759..1768
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1773..1775,1776..1778,1779..2291)
                     /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             4677..5411
                     /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             5469..6203
                     /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"
     intron          6352..6484
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     6467..6486
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    6493..7081
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6546..6566)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6964..6975)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7108..7125)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7114..7225
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7321..7341)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7321..7339)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7360..7376)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7478..7497)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7543..7598
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7597..7616)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7959..7975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7983..7999
                     /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(8007..8013,8014..8031,8032..8037))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8052..8073
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8132..8327
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8178..8313
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8333..8467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8370..8389)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8424..8443
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8535..8552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8706..9294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8786..8805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9465..10256,10257..10325))
                     /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     10088..10107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10326..10430)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccrgggggg gggggggggg gggggggggg
      541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccgc
     1741 gggcccagct tgccgccacc atggcggccg caatggtgag caagggcgag gagctgttca
     1801 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1861 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1921 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgtcc tggggcgtgc
     1981 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2041 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2101 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2161 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac tttagcgaca
     2221 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     2281 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2341 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     2401 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgcccgat
     2461 atcagcccgt gggaactcat gagatggagg aggagttggc cgaagctgtg gccctgctct
     2521 cccagcgggg gcctgacgcc ctgctcactg tggcacttcg aaagccccca ggtcagcgca
     2581 cggatgaaga gctggacctc atctttgagg agctgctgca catcaaggct gtggcccacc
     2641 tctccaactc ggtgaagcga gaattagcgg ctgttctgct ctttgaacca cacagcaagg
     2701 cagggaccgt gttgttcagc cagggggaca agggcacttc gtggtacatt atctggaagg
     2761 gatctgtcaa cgtggtgacc catggcaagg ggctggtcac caccctgcat gagggagatg
     2821 attttggaga gctggctctg gtgaatgatg caccccgggc agccaccatc atcctgcgag
     2881 aagacaactg tcatttcctg cgtgtggaca agcaggactt caaccgtatc atcaaggatg
     2941 tggaggcaaa gaccatgcgg ctggaagaac atggcaaagt ggtgctggtg ctggagagag
     3001 cctctcaggg cgccggccct tcccgacccc caaccccagg caggaaccgg tatacagtga
     3061 tgtctggcac cccagagaag atcctagagc ttctgttgga ggccatggga ccagattcca
     3121 gtgctcatga cccaacagag acattcctca gcgacttcct cctgacccac agggtcttca
     3181 tgcccagcgc ccaactctgc gctgcccttc tgcaccactt ccatgtggag cctgcgggtg
     3241 gcagcgagca ggagcgcagc acctacgtct gcaacaagag gcagcagatc ttgcggctgg
     3301 tcagccagtg ggtggccctg tatggctcca tgctccacac tgaccctgtg gccaccagct
     3361 tcctccagaa actctcagac ctggtgggca gggacacccg actcagcaac ctgctgaggg
     3421 agcagtggcc agagaggcgg cgatgccaca ggttggagaa tggctgtggg aatgcatctc
     3481 ctcagatgaa ggcccggaac ttgcctgttt ggctccccaa ccaggacgag ccccttcctg
     3541 gcagcagctg tgccatccaa gttggggata aagtccccta tgacatctgc cggccagacc
     3601 actcagtgtt gaccctgcag ctgcctgtga cagcctccgt gagagaggtg atggcagcgt
     3661 tggcccagga ggatggctgg accaaggggc aggtgctggt gaaggtcaat tctgcaggtg
     3721 atgccattgg cctgcagcca gatgcccgtg gtgtggccac atctctgggg ctcaatgagc
     3781 gtctctttgt tgtcaaccca caggaagcgc atgagctgat cccacaccct gaccagctgg
     3841 ggcccactgt gggctctgct gaggggctgg acctggtgag tgccaaggac ctggcaggcc
     3901 agctgacgga ccacgactgg agcctcttca acagtatcca ccaggtggag ctgatccact
     3961 atgtgctggg cccccagcat ctgcgggatg tcaccaccgc caacctggag cgcttcatgc
     4021 gccgcttcaa tgagctgcag tactgggtgg ccaccgagct gtgtctctgc cccgtgcccg
     4081 gcccccgggc ccagctgctc aggaagttca ttaagctggc ggcccacctc aaggagcaga
     4141 agaatctcaa ttccttcttt gccgtcatgt ttggcctcag caactcggcc atcagccgcc
     4201 tagcccacac ctgggagcgg ctgccccaca aagtccggaa gctgtactcc gccctcgaga
     4261 ggctgctgga tccctcatgg aaccaccggg tataccgact ggccctcgcc aagctctccc
     4321 ctcctgtcat ccccttcatg ccccttcttg cagcagacat ggccgccatt catgagggaa
     4381 accacacact agtggagaat ctcatcaact ttgagaagat gagaatgatg gccagagccg
     4441 cgcggatgct gcaccactgc cgaagccaca accctgtgcc tctctcacca ctcagaagcc
     4501 gagtttccca cctccacgag gacagccagg tggcgaggat ttccacatgc tcggagcagt
     4561 ccctgagcac ccggagtcca gccagcacct gggcttatgt ccagcagctg aaggtcattg
     4621 acaaccagcg ggaactctcc cgcctctccc gagagctgga gccagctagc gagctcatgg
     4681 acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     4741 tgctgctgcc cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg
     4801 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     4861 tggacgagct gtacaagggt ggcagcggtg gcatggtgag caagggcgag gagctgttca
     4921 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4981 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctgatctgca
     5041 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc tacggcctgc
     5101 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     5161 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     5221 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     5281 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     5341 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     5401 acaacatcga gccgaatttt gtcttcctga tcggcgccgc aggaatactc ttcgtatcta
     5461 gcgagctcat ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     5521 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca
     5581 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     5641 tcactctcgg catggacgag ctgtacaagg gtggcagcgg tggcatggtg agcaagggcg
     5701 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     5761 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     5821 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     5881 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     5941 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     6001 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     6061 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     6121 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     6181 tcaagatccg ccacaacatc gagccgaatt ccctcgaggt taacgcggcg cgcctctaat
     6241 ctaggggatc caccggaagc cacctgcatg cggccgcatg atcagctgga tgcatcgatc
     6301 acgcgtaccg gtgctcgagg taccgatgaa tagctaaggt cgaggccgca ggtaagtatc
     6361 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
     6421 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
     6481 acaggtgtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     6541 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6601 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     6661 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     6721 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     6781 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     6841 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     6901 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     6961 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     7021 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     7081 cctggaattc gagctcggta cgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7141 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7201 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7261 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagccca gcttttgttc
     7321 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7381 aaattgttat ccgctaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg
     7441 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
     7501 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     7561 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     7621 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     7681 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     7741 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     7801 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     7861 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     7921 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     7981 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     8041 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     8101 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     8161 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     8221 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     8281 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     8341 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     8401 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     8461 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     8521 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     8581 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     8641 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     8701 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     8761 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     8821 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     8881 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     8941 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     9001 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     9061 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     9121 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     9181 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     9241 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     9301 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     9361 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     9421 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     9481 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     9541 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     9601 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     9661 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     9721 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     9781 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     9841 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     9901 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     9961 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    10021 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    10081 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    10141 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    10201 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    10261 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    10321 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    10381 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    10441 cgaaaagtgc cacctg
//
