LOCUS       sequence-353306-       10021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tet-on expression of an inward-rectifier potassium channel Kir2.1 
            carrying an CFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10021)
  AUTHORS   Tian H, Davis HC, Wong-Campos JD, Park P, Fan LZ, Gmeiner B, Begum 
            S, Werley CA, Borja GB, Upadhyay H, Shah H, Jacques J, Qi Y, Parot 
            V, Deisseroth K, Cohen AE
  TITLE     Video-based pooled screening yields improved far-red genetically 
            encoded voltage indicators.
  JOURNAL   Nat Methods. 2023 Jul;20(7):1082-1094. doi: 
            10.1038/s41592-022-01743-5. Epub 2023 Jan 9.
  PUBMED    36624211
REFERENCE   2  (bases 1 to 10021)
  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"; doi: 
            "10.1038/s41592-022-01743-5"; journalName: "Nat Methods"; date: 
            "2023-07"; volume: "20"; issue: "7"; pages: "1082-1094"
COMMENT     Sequence Label: TDG004_pLenti_CMVtight_Kir2.1_CFP
FEATURES             Location/Qualifiers
     source          1..10021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(131..199,200..991)
                     /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(349..368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1162..1750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1651..1670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1904..1921
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2074..2079,2080..2097,2098..2104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2112..2128
                     /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     2117..2139
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2136..2152
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2136..2152
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2171..2191
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2173..2191
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2219..2445
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2446..2626
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2673..2798
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3291..3524
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3709..3753
                     /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    4020..4137
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4159..4473
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    4167..4185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4203..4221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4238..4256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4274..4292
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4310..4328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4345..4363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4381..4399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4446..4470
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    4551..4575
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4623..4643
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      4643..4652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5945..5947,5948..5950,5951..6664)
                     /codon_start=1
                     /product="monomeric super enhanced variant of CFP (Matsuda 
                     et al., 2008)"
                     /label=mseCFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(5945..5947,5948..5950,5951..6664)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             join(5945..5947,5948..5950,5951..6664)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(5990..6011)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6251..6270)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6598..6619
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6665..6689)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6720..7308
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6773..6793)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7191..7202)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        7351..7850
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7386..7409)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7783..7802
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7864..8667
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7927..7946)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8537..8556
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             8758..8991
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9063..9184
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9100..9119)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9154..9173
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9224..9359
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9379..9398)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9380..9398)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9408..9425)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9408..9424)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9417..9439)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9566..9994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9653..9672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9863..9884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     2161 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     2221 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     2281 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     2341 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     2401 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     2461 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2521 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2581 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     2641 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     2701 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     2761 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     2821 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     2881 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     2941 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     3001 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     3061 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     3121 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     3181 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     3241 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     3301 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     3361 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     3421 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     3481 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     3541 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     3601 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     3661 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     3721 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     3781 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     3841 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     3901 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     3961 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttaact
     4021 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     4081 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttatc
     4141 gatcacgaga ctagcctcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
     4201 actccctatc agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt
     4261 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
     4321 gatagagaac gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac
     4381 tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga ggcctatata
     4441 agcagagctc gtttagtgaa ccgtcagatc gcctggagaa ttcgagctcg gtacccgggg
     4501 atccaccggt tcgtcgacta gtccagtgtg gtggaattct gcagatatca acaagtttgt
     4561 acaaaaaagc aggctttaaa ggaaccaatt cagtcgactg gatctcgagc tcaagcttat
     4621 gccaactttg tatacaaaag ttgccgccat gggcagtgtg agaaccaacc gctacagcat
     4681 cgtctcttcg gaggaagatg gcatgaagct ggccactatg gcagttgcca atggctttgg
     4741 gaatggcaag agtaaagtcc atacccgaca acagtgcagg agccgctttg tgaagaaaga
     4801 tggtcattgc aatgttcagt ttatcaacgt gggtgagaag ggacagaggt acctggcaga
     4861 catctttact acctgtgtcg acatccgctg gaggtggatg ctggttatct tctgtcttgc
     4921 cttcgtgctc tcctggctgt tctttggctg tgtgttttgg ttgatagccc tgctccatgg
     4981 ggatctagat acttctaaag tgagcaaagc atgcgtgtca gaggtcaaca gcttcacggc
     5041 tgccttcctc ttctccatcg agacccagac aaccattggc tatggtttca ggtgtgtgac
     5101 agacgagtgc ccaattgctg tcttcatggt ggtattccag tcaatcgtag gctgcatcat
     5161 tgacgccttc atcattggtg cagtcatggc gaagatggca aagccaaaga agagaaatga
     5221 gactcttgtc ttcagtcaca atgctgtgat tgccatgagg gatggcaaac tctgcttgat
     5281 gtggagagtg ggtaaccttc gaaagagcca ccttgtggaa gctcatgtcc gggcacagct
     5341 tctcaaatct aggatcactt cagaagggga gtatatccct ttggaccaga tagacatcaa
     5401 tgttggtttt gatagtggaa ttgaccgcat atttctagtg tcccccatca ctatcgttca
     5461 cgaaatagat gaagacagcc ctttatatga cttgagtaag caggacattg acaatgcaga
     5521 ctttgaaatt gttgtcatac tggaaggcat ggtggaggcg actgccatga caactcaatg
     5581 ccggagttcg tatctggcca atgaaattct ctggggtcac cgctatgagc cagtgctctt
     5641 tgaagagaaa cactactata aagtagacta ttcaagattc cataagactt atgaagtacc
     5701 taacaccccc ctttgtagtg ccagagactt agcagagaag aaatacatcc tttcaaatgc
     5761 aaattcattt tgctatgaaa atgaagttgc cctaacaagc aaagaggaag aggaggatag
     5821 tgagaacgga gtcccagaga gcacaagcac agactcacct cctggcatag atctccacaa
     5881 ccaggcaagc gtacctctag agcccaggcc cttaaggcga gaatcggaga tagggccggc
     5941 catgatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     6001 ggacggcgac gtaaacggcc acaggttcag cgtgtccggc gagggcgagg gcgatgccac
     6061 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     6121 caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc ccgaccacat
     6181 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgtaccat
     6241 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     6301 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     6361 gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg acaagcagaa
     6421 gaacggcatc aaggcccact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     6481 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     6541 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     6601 ggtcctgctg gagttcgtga ccgccgccgg gatcactcac ggcatggacg agctgtacaa
     6661 gtgaacccag ctttcttgta caaagtggtt gatatccagc acagtggcgg ccgctcgaca
     6721 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     6781 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     6841 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     6901 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     6961 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     7021 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     7081 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     7141 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     7201 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     7261 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattctgc
     7321 agatatccgg actagtgatc taattctacc gggtagggga ggcgcttttc ccaaggcagt
     7381 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     7441 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     7501 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     7561 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     7621 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     7681 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     7741 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     7801 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     7861 aatatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     7921 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     7981 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     8041 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     8101 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     8161 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     8221 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     8281 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     8341 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     8401 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     8461 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     8521 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     8581 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     8641 tatcgccttc ttgacgagtt cttctgaggg gatccgtcga gggcccggta cctttaagac
     8701 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     8761 aagggctagc tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     8821 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     8881 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     8941 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     9001 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     9061 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     9121 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     9181 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     9241 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     9301 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     9361 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     9421 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     9481 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     9541 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     9601 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     9661 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     9721 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     9781 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     9841 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     9901 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     9961 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
    10021 t
//
