pDisplay-nLightG-IRES-mcherry-CAAX

LOCUS       pDisplay-nLightG-IRES-mcherry-CAAX              8495 bp DNA     circular SYN 23-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8495)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 29, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11..627
                     /label=CMV_immearly_promoter
     misc_feature    90..377
                     /label=CAG_enhancer
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    638..656
                     /label=T7_promoter
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     696..754
                     /label=F-pdisplay-EcoRI-gb-nLight
     CDS             737..2686
                     /codon_start=1
                     /label=nLightG
                     /translation="MKTIIALSYIFCLVFADYKDDDDAVFLSGNASDSSNCTHPPPPVN
                     ISKAILLGVILGGLILFGVLGNILVILSVACHRHLHSVTHYYIVNLAVADLLLTSTVLP
                     FSAIFEILGYWAFGRVFCNIWAAVDVLCCTASIMGLCLISIDRYIGVSYPLRYPTIVTQ
                     KRGLMALLCVWALSLVISIGPLFGWRQPAPEDETICQINEEPGYVLFSALGSFYVPLTI
                     ILVMYCRVYRIAQKQLSSLINVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTP
                     IGDGPVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMV
                     SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTL
                     VTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTL
                     VNRIELKGIDFKEDGNILGHKLEYNNHDQLKRETKVAKTLGIVVGCFVLCWLPFFLVMP
                     IGSFFPDFRPSETVFKIAFWLGYLNSCINPIIYPCSSQEFKKAFQNVLRIQCLRRKQSS
                     KHALGYTLHPPSHALEGQYRNLVRIPVGSGETFCKISKTDGVCEWKFFSSLPRGSARIT
                     VPKDQSACTTARDF"
     misc_feature    737..2221
                     /label=nLightG-Nter_cpEGFP
     primer_bind     complement(2200..2246)
                     /label=R-nLight-Cter-gb-nLightG-Nter
     misc_feature    2222..2686
                     /label=nLight-Cter
     primer_bind     2222..2240
                     /label=F-nLight-Cter
     primer_bind     complement(2669..2717)
                     /label=R-IRES-NheI-gb-nLight-Cter
     misc_feature    2693..3276
                     /label=IRES
     misc_feature    2694..3279
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3280..4038
                     /codon_start=1
                     /label=ORF2
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYKDGKKKKKKSKTKCVIM"
     CDS             3280..3987
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    3988..4035
                     /label=CAAX
     CDS             4045..4074
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    4097..4102
                     /label=SDS
     polyA_signal    4252..4476
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4593..5212)
                     /direction=LEFT
                     /label=pBR322_origin
     rep_origin      complement(4608..5196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    5525..5572
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             5753..6289
                     /codon_start=1
                     /label=ORF frame 2
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEAV
                     SPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAIGHDEILAVG
                     HARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQPPH
                     CISHDGYFLGRSKVR"
     CDS             complement(5804..6598)
                     /codon_start=1
                     /label=ORF frame 3
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTYGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(6633..6986)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6647..6778
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    6661..6666
                     /label=SDS
     rep_origin      complement(6701..6778)
                     /direction=LEFT
                     /label=SV40_origin
     gene            complement(7046..7906)
                     /label=Ampicillin
     CDS             complement(7046..7906)
                     /codon_start=1
                     /label=ORF2
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7907..8011)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(7948..7976)
                     /label=AmpR_promoter
     rep_origin      8038..8466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      8055..8361
                     /label=f1_origin
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccctgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg
      721 cttggggata tccaccatga agacgatcat cgccctgagc tacatcttct gcctggtgtt
      781 cgccgactac aaggacgatg atgacgcagt ctttctcagt gggaacgcta gtgactcttc
      841 aaattgtacc cacccaccac ctcctgttaa tatcagcaag gctattttgc tgggtgtcat
      901 cttgggaggc ctgattctgt tcggagtgct tggtaacatt ctcgtaattc tctcagtagc
      961 gtgtcaccgc catctccaca gcgttaccca ctactatatt gtgaacctcg ctgtagcaga
     1021 cctgcttctc acgagtacgg tactgccatt ttccgctatc ttcgaaattt tggggtattg
     1081 ggcctttgga cgggtttttt gtaacatctg ggccgctgtg gacgtattgt gttgtacggc
     1141 atcaatcatg ggcctgtgcc ttatctcaat agatcgatac attggagtgt cttatcctct
     1201 gaggtatccg accatagtaa cccaaaaacg agggttgatg gctctccttt gtgtctgggc
     1261 tctttccttg gttatatcca ttgggccctt gttcggctgg cgccagcccg ccccggagga
     1321 cgaaacaata tgtcagatca acgaagagcc tggatacgtc cttttcagcg ccctcggatc
     1381 tttttatgtc ccgttgacaa tcatcctggt catgtattgc agagtttata ggattgctca
     1441 gaaacagctg agctcactca ttaacgtcta tatcaaggcc gacaagcaga agaacggcat
     1501 caaggcgaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca
     1561 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1621 gagcgtgcag tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1681 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac
     1741 cggagggagc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1801 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga
     1861 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1921 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1981 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg
     2041 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2101 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2161 cctggggcac aagctggagt acaacaatca tgaccaactg aaaagagaaa ctaaagtcgc
     2221 taagacgctc gggatagttg ttggttgttt cgtactctgc tggttgccct ttttcttggt
     2281 gatgccgata ggtagctttt ttcccgactt ccgaccgagt gagacagtat tcaaaatagc
     2341 tttttggctt ggttatttga actcctgtat taatcccatt atatatccct gttcatctca
     2401 agagtttaaa aaggcttttc aaaacgtgtt gcgcatacaa tgcctgcgca gaaagcagtc
     2461 ttctaagcac gcactcggtt atacgctcca ccccccatcc catgctctcg agggacagta
     2521 tagaaacctc gtgcgaatac ctgtgggtag tggcgaaaca ttttgcaaaa tcagtaaaac
     2581 agacggggta tgtgaatgga aatttttttc atccttgcca cgcggatctg ccagaataac
     2641 agttcctaag gaccagtccg cttgcacaac tgcccgggac ttttaagcta gcgcccctct
     2701 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     2761 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     2821 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     2881 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     2941 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     3001 caacgccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     3061 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     3121 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     3181 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3241 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     3301 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     3361 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     3421 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     3481 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     3541 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3601 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     3661 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     3721 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     3781 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     3841 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     3901 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     3961 accggcggca tggacgagct gtacaaggat ggtaaaaaga agaaaaagaa gtcaaagaca
     4021 aagtgtgtaa ttatgtaagt cgacgaacaa aaactcatct cagaagagga tctgaatgct
     4081 gtgggccagg acacgcagga ggtcatcgtg gtgccacact ccttgccctt taaggtggtg
     4141 gtgatctcag ccatcctggc cctggtggtg ctcaccatca tctcccttat catcctcatc
     4201 atgctttggc agaagaagcc acgttaggcg gccgctcgag atcagcctcg actgtgcctt
     4261 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4321 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4381 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4441 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagtg
     4501 gcggtaatac ggttatccac agaatcgggg gataacgcag gaaagaacat gtgagcaaaa
     4561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4921 tccaacccgg taagacgcga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5341 atatatgagt aacctgaggc tatggcaggg cctgccgccc cgacgttggc tgcgagccct
     5401 gggccttcac ccgaacttgg ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg
     5461 ccccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt
     5521 ttatgaacaa acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg
     5581 cgggttcctt ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg
     5641 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     5701 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     5761 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     5821 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     5881 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     5941 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     6001 tttccaccat gatattcggc aagcaggcat cgccataggt cacgacgaga tcctcgccgt
     6061 cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     6121 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     6181 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     6241 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     6301 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     6361 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcttgca
     6421 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     6481 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     6541 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     6601 gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa agcctaggcc tccaaaaaag
     6661 cctcctcact acttctggaa tagctcagag gccgggcctc ggcctctgca taaataaaaa
     6721 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcggaatgg
     6781 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     6841 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     6901 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     6961 tccacagctg gttctttccg cctcaggact cttccttttt caataaatca atctaaagta
     7021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     8041 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     8101 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     8161 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     8221 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     8281 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     8341 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     8401 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     8461 cgaattttaa caaaatatta acgcttacaa tttac
//