LOCUS       sequence-236937-        8487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces equimolar expression of yeast codon-optimized tagRFPT and 
            mTq2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8487)
  AUTHORS   Botman D, de Groot DH, Schmidt P, Goedhart J, Teusink B
  TITLE     In vivo characterisation of fluorescent proteins in budding yeast.
  JOURNAL   Sci Rep. 2019 Feb 19;9(1):2234. doi: 10.1038/s41598-019-38913-z.
  PUBMED    30783202
REFERENCE   2  (bases 1 to 8487)
  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: "Sci Rep."; 
            date: "2019-02-19"; volume: "9(1):2234. doi"; pages: " 
            10.1038/s41598-019-38913-z"
COMMENT     Sequence Label: pDRF1-GW yotagRFPT-T2A-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..8487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    800..824
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      829..838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1573..1626
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      1642..1651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1648..1650,1651..1653,1654..2367)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1693..1714)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1954..1973)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2301..2322
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2377..2401)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2463..2482)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2464..2482)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2630..2817
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2685..2709)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2829..2846)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2829..2845)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2838..2860)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3281..4627
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(4738..5541)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(5542..5757)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5782..5800)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5868..5972
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5973..6041,6042..6833)
                     /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(6191..6210)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7004..7592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7493..7512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7746..7763
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7880..7901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7916..7921,7922..7939,7940..7946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7954..7970
                     /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     7959..7981
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7978..7994
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7978..7994
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(8024..8479)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8161..8182)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8373..8392
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 caagcttcct gaaacggaga aacataaaca ggcattgctg ggatcaccca tacatcactc
       61 tgttttgcct gaccttttcc ggtaatttga aaacaaaccc ggtctcgaag cggagatccg
      121 gcgataatta ccgcagaaat aaacccatac acgagacgta gaaccagccg cacatggccg
      181 gagaaactcc tgcgagaatt tcgtaaactc gcgcgcattg catctgtatt tcctaatgcg
      241 gcacttccag gcctcgatcg agaccgttta tccattgctt ttttgttgtc tttttccctc
      301 gttcacagaa agtctgaaga agctatagta gaactatgag ctttttttgt ttctgttttc
      361 cttttttttt tttttacctc tgtggaaatt gttactctca cactctttag ttcgtttgtt
      421 tgttttgttt attccaatta tgaccggtga cgaaacgtgg tcgatggtgg gtaccgctta
      481 tgctcccctc cattagtttc gattatataa aaaggccaaa tattgtatta ttttcaaatg
      541 tcctatcatt atcgtctaac atctaatttc tcttaaattt tttctctttc tttcctataa
      601 caccaatagt gaaaatcttt ttttcttcta tatctacaaa aacttttttt ttctatcaac
      661 ctcgttgata aattttttct ttaacaatcg ttaataatta attaattgga aaataaccat
      721 tttttctctc ttttatacac acattcaaaa gaaagaaaaa aaatataccc cagcctcgac
      781 tagtgaattc gcggccatca caagtttgta caaaaaagca ggctgctagc caccatggta
      841 tctaaaggtg aagagttgat aaaggaaaat atgcacatga aactctatat ggaaggcaca
      901 gtgaataacc atcattttaa gtgtacttca gaaggagaag gtaagcctta tgaggggaca
      961 caaactatga gaattaaagt tgttgaagga ggtccactac cattcgcttt tgacatcctg
     1021 gcaacgagct tcatgtatgg ctccagaaca ttcataaacc atacacaagg tattccagat
     1081 ttcttcaaac aaagttttcc agaaggcttt acttgggaga gagttactac ctacgaggac
     1141 gggggtgttt taactgcgac acaggataca tcattacaag atggctgttt gatctacaac
     1201 gtaaaaatca gaggtgtcaa ttttccatct aacggacctg ttatgcagaa aaagaccttg
     1261 ggttgggagg ctaatactga aatgctatac ccagcagatg gaggattgga aggtaggaca
     1321 gatatggcat tgaaacttgt aggaggcggt cacctaattt gcaactttaa gacaacctac
     1381 agaagcaaaa agcctgctaa aaatcttaaa atgccagggg tgtactacgt ggatcataga
     1441 cttgaacgta tcaaggaagc tgacaaagag acttacgttg aacaacacga agtcgccgtc
     1501 gccagatact gtgacttacc ttcaaagtta ggtcataaac tgaatggtat ggatgaattg
     1561 tataagtccg gagagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     1621 ggccctgggg atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     1681 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     1741 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1801 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgtcctgggg cgtgcagtgc
     1861 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     1921 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1981 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2041 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actactttag cgacaacgtc
     2101 tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac
     2161 atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2221 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccaagct gagcaaagac
     2281 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     2341 ctcggcatgg acgagctgta caagtaaagc ggccgcaccc agctttcttg tacaaagtgg
     2401 tgtagttaat tcatgatggc cgctgcaggt cgacctcgag ggggggcccg gtacccaatt
     2461 cgccctatag tgagtcgtat tacgcgcgga tccagctttg gacttcttcg ccagaggttt
     2521 ggtcaagtct ccaatcaagg ttgtcggctt gtctaccttg ccagaaattt acgaaaagat
     2581 ggaaaagggt caaatcgttg gtagatacgt tgttgacact tctaaataag cgaatttctt
     2641 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     2701 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     2761 caggttgctt tctcaggtat agcatgaggt cgctcttatt gaccacacct ctaccggcat
     2821 gccaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2881 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     2941 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     3001 tttctcctta cgcatctgtg cggtatttca caccgcataa tcggatcgta cttgttaccc
     3061 atcattgaat tttgaacatc cgaacctggg agttttccct gaaacagata gtatatttga
     3121 acctgtataa taatatatag tctagcgctt tacggaagac aatgtatgta tttcggttcc
     3181 tggagaaact attgcatcta ttgcataggt aatcttgcac gtcgcatccc cggttcattt
     3241 tctgcgtttc catcttgcac ttcaatagca tatctttgtt aacgaagcat ctgtgcttca
     3301 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     3361 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     3421 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     3481 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     3541 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     3601 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     3661 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     3721 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     3781 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     3841 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     3901 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     3961 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     4021 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     4081 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     4141 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     4201 gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     4261 tagctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt
     4321 ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg
     4381 cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa
     4441 tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata
     4501 ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta
     4561 tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata
     4621 ttggatcgat ccgatgataa gctgtcaaac atgagaattg ggtaataact gatataatta
     4681 aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata cagtttttta
     4741 gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct gcttttctgt aacgttcacc
     4801 ctctacctta gcatcccttc cctttgcaaa tagtcctctt ccaacaataa taatgtcaga
     4861 tcctgtagag accacatcat ccacggttct atactgttga cccaatgcgt ctcccttgtc
     4921 atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa ccttcatctc ttccacccat
     4981 gtctctttga gcaataaagc cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt
     5041 acccttagta tattctccag tagataggga gcccttgcat gacaattctg ctaacatcaa
     5101 aaggcctcta ggttcctttg ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc
     5161 tgggcccacc acaccgtgtg cattcgtaat gtctgcccat tctgctattc tgtatacacc
     5221 cgcagagtac tgcaatttga ctgtattacc aatgtcagca aattttctgt cttcgaagag
     5281 taaaaaattg tacttggcgg ataatgcctt tagcggctta actgtgccct ccatggaaaa
     5341 atcagtcaag atatccacat gtgtttttag taaacaaatt ttgggaccta atgcttcaac
     5401 taactccagt aattccttgg tggtacgaac atccaatgaa gcacacaagt ttgtttgctt
     5461 ttcgtgcatg atattaaata gcttggcagc aacaggacta ggatgagtag cagcacgttc
     5521 cttatatgta gctttcgaca tgatttatct tcgtttcctg catgtttttg ttctgtgcag
     5581 ttgggttaag aatactgggc aatttcatgt ttcttcaaca ctacatatgc gtatatatac
     5641 caatctaagt ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa
     5701 aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag
     5761 ctaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5821 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5881 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5941 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6001 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6061 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6121 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6181 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6241 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6301 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6361 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6421 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6481 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6541 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6601 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6661 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6721 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6781 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6841 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6901 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6961 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7021 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7081 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7141 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7201 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7261 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7321 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7381 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7441 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7501 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7561 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7621 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7681 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7741 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7801 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     7861 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     7921 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     7981 gaaacagcta tgaccatgat tacgccaagc ttaccgcatc aggaaattgt aagcgttaat
     8041 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     8101 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     8161 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     8221 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     8281 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     8341 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     8401 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     8461 gcgccgctac agggcgcgtc cattcgc
//
