LOCUS       sequence-475691-        9205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpress Sc FLAG-RFC1 in yeast (integrated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9205)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 9205)
  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"
COMMENT     Sequence Label: Sc FLAG-RFC1-pRS405/GAL-L
FEATURES             Location/Qualifiers
     source          1..9205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..27)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7..25)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(46..62)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(46..62)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(59..81)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    70..86
                     /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(94..100,101..118,119..124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    139..160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(277..294)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(448..1036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(528..547)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1207..1998,1999..2067))
                     /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     1830..1849
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2068..2172)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2240..2258
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2296..2318)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2418..2437
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2931..4025)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4026..4433)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(4734..5189)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4871..4892)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5083..5102
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5315..5337
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5329..5346
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5330..5346
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5356..5375
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5356..5374
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5399..5415
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5400..5416
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5444..6108
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    5660..5777
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6072..6095
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             6127..6192
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(8917..8935)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8931..8978
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(9153..9158,9159..9175,9176..9181))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
       61 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      121 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      181 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      241 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      301 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      361 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      421 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      481 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      541 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      601 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      661 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      721 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      781 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      841 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
      901 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
      961 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     2221 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     2281 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2341 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2401 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatcgacta
     2461 cgtcgttaag gccgtttctg acagagtaaa attcttgagg gaactttcac cattatggga
     2521 aatggttcaa gaaggtattg acttaaactc catcaaatgg tcaggtcatt gagtgttttt
     2581 tatttgttgt attttttttt ttttagagaa aatcctccaa tatataaatt aggaatcata
     2641 gtttcatgat tttctgttac acctaacttt ttgtgtggtg ccctcctcct tgtcaatatt
     2701 aatgttaaag tgcaattctt tttccttatc acgttgagcc attagtatca atttgcttac
     2761 ctgtattcct ttacatcctc ctttttctcc ttcttgataa atgtatgtag attgcgtata
     2821 tagtttcgtc taccctatga acatattcca ttttgtaatt tcgtgtcgtt tctattatga
     2881 atttcattta taaagtttat gtacaaatat cataaaaaaa gagaatcttt ttaagcaagg
     2941 attttcttaa cttcttcggc gacagcatca ccgacttcgg tggtactgtt ggaaccacct
     3001 aaatcaccag ttctgatacc tgcatccaaa acctttttaa ctgcatcttc aatggcctta
     3061 ccttcttcag gcaagttcaa tgacaatttc aacatcattg cagcagacaa gatagtggcg
     3121 atagggttga ccttattctt tggcaaatct ggagcagaac cgtggcatgg ttcgtacaaa
     3181 ccaaatgcgg tgttcttgtc tggcaaagag gccaaggacg cagatggcaa caaacccaag
     3241 gaacctggga taacggaggc ttcatcggag atgatatcac caaacatgtt gctggtgatt
     3301 ataataccat ttaggtgggt tgggttctta actaggatca tggcggcaga atcaatcaat
     3361 tgatgttgaa ccttcaatgt agggaattcg ttcttgatgg tttcctccac agtttttctc
     3421 cataatcttg aagaggccaa aacattagct ttatccaagg accaaatagg caatggtggc
     3481 tcatgttgta gggccatgaa agcggccatt cttgtgattc tttgcacttc tggaacggtg
     3541 tattgttcac tatcccaagc gacaccatca ccatcgtctt cctttctctt accaaagtaa
     3601 atacctccca ctaattctct gacaacaacg aagtcagtac ctttagcaaa ttgtggcttg
     3661 attggagata agtctaaaag agagtcggat gcaaagttac atggtcttaa gttggcgtac
     3721 aattgaagtt ctttacggat ttttagtaaa ccttgttcag gtctaacact accggtaccc
     3781 catttaggac cacccacagc acctaacaaa acggcatcag ccttcttgga ggcttccagc
     3841 gcctcatctg gaagtggaac acctgtagca tcgatagcag caccaccaat taaatgattt
     3901 tcgaaatcga acttgacatt ggaacgaaca tcagaaatag ctttaagaac cttaatggct
     3961 tcggctgtga tttcttgacc aacgtggtca cctggcaaaa cgacgatctt cttaggggca
     4021 gacattagaa tggtatatcc ttgaaatata tatatatata ttgctgaaat gtaaaaggta
     4081 agaaaagtta gaaagtaaga cgattgctaa ccacctattg gaaaaaacaa taggtcctta
     4141 aataatattg tcaacttcaa gtattgtgat gcaagcattt agtcatgaac gcttctctat
     4201 tctatatgaa aagccggttc cggcgctctc acctttcctt tttctcccaa tttttcagtt
     4261 gaaaaaggta tatgcgtcag gcgacctctg aaattaacaa aaaatttcca gtcatcgaat
     4321 ttgattctgt gcgatagcgc ccctgtgtgt tctcgttatg ttgaggaaaa aaataatggt
     4381 tgctaagaga ttcgaactct tgcatcttac gatacctgag tattcccaca gttaactgcg
     4441 gtcaagatat ttcttgaatc aggcgcctta gaccgctcgg ccaaacaacc aattacttgt
     4501 tgagaaatag agtataatta tcctataaat ataacgtttt tgaacacaca tgaacaagga
     4561 agtacaggac aattgatttt gaagagaatg tggattttga tgtaattgtt gggattccat
     4621 ttttaataag gcaataatat taggtatgta gatatactag aagttctcct cgaccggtcg
     4681 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     4741 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4801 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4861 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4921 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4981 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5041 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5101 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5161 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     5221 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     5281 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     5341 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gccccccctc
     5401 gaggtcgacg gtatcgataa gcttgatatc gaattccttg aattttcaaa aattcttact
     5461 ttttttttgg atggacgcaa agaagtttaa taatcatatt acatggcatt accaccatat
     5521 acatatccat atacatatcc atatctaatc ttacttatat gttgtggaaa tgtaaagagc
     5581 cccattatct tagcctaaaa aaaccttctc tttggaactt tcagtaatac gcttaactgc
     5641 tcattgctat attgaagtac ggattagaag ccgccgagcg ggtgacagcc ctccgaagga
     5701 agactctcct ccgtgcgtcc tcgtcttcac cggtcgcgtt cctgaaacgc agatgtgcct
     5761 cgcgccgcac tgctccgaac aataaagatt ctacaatact agcttttatg gttatgaaga
     5821 ggaaaaattg gcagtaacct ggccccacaa accttcaaat gaacgaatca aattaacaac
     5881 cataggatga taatgcgatt agttttttag ccttatttct ggggtaatta atcagcgaag
     5941 cgatgatttt tgatctatta acagatatat aaatgcaaaa actgcataac cactttaact
     6001 aatactttca acattttcgg tttgtattac ttcttattca aatgtaataa aagtatcaac
     6061 aaaaaattgt taatatacct ctatacttta acgtcaagga gaaaaaaccc cggatccact
     6121 agtatggact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     6181 gacgatgaca agtccatggt caatatttct gatttctttg gtaaaaataa gaaatccgta
     6241 agatcgtcaa catccagacc tacgagacag gtgggttcgt ctaaaccaga agttatcgac
     6301 ttagatactg aatctgacca agaatcaacc aataaaacac caaagaagat gcctgtaagt
     6361 aatgtaattg atgtatcaga gacacctgaa ggagaaaaaa agctacctct tcccgctaag
     6421 aggaaggcct cttcaccaac tgtaaagcca gcaagttcaa aaaaaaccaa accctcctct
     6481 aaaagttcag actcggcttc gaatattacg gctcaagatg tactagataa gattccatcc
     6541 ttggatctgt caaatgttca tgtgaaagag aatgctaaat ttgatttcaa aagcgcaaat
     6601 tcaaatgcgg atccagatga aattgtttct gaaataggta gttttccaga aggaaagcca
     6661 aattgtttat taggtctaac aattgtcttc acaggtgttc tgccaacctt ggagcgtggt
     6721 gcatcggaag ctttagcgaa aagatacgga gcaagggtta ctaaatcaat ttcaagcaaa
     6781 acttctgtag tagtcttagg tgatgaagca gggccaaaga aactagaaaa aatcaagcaa
     6841 ttgaaaatca aagccataga cgaagaaggc tttaagcaat taattgctgg gatgcctgct
     6901 gaaggtggtg acggggaagc tgccgaaaaa gcacgtcgaa aattagaaga acagcataat
     6961 attgctacca aagaagcaga attgcttgtt aagaaagaag aggaaaggtc gaagaagctt
     7021 gcagccacta gagtttctgg tggccatctg gagagagaca atgtggttag ggaagaagat
     7081 aaattgtgga cagtaaagta cgcaccaacg aatctacaac aggtctgtgg taacaaagga
     7141 agtgtaatga aattaaaaaa ctggttggct aattgggaaa attcaaagaa aaatagtttt
     7201 aaacatgctg gtaaagatgg ctctggtgtt ttcagggctg caatgttata cggtcctcct
     7261 ggtattggga agacaactgc tgctcattta gttgcacaag agcttgggta tgacattctt
     7321 gaacaaaacg cttctgatgt tcgctctaaa actttactta atgccggtgt taaaaacgct
     7381 cttgataaca tgtctgtagt gggatatttc aaacacaacg aagaagctca aaatttgaac
     7441 gggaaacatt ttgtcattat catggatgaa gttgatggta tgagcggagg cgataggggg
     7501 ggagttggcc agctggcaca gttttgtcgt aagacgtcaa ctccattgat tctaatttgt
     7561 aatgaacgta atttaccaaa aatgagacca ttcgatagag tatgtcttga tatccaattt
     7621 agaagacctg atgctaacag tatcaaatca agattaatga caattgcgat cagagaaaag
     7681 ttcaaacttg atccaaatgt cattgatagg ttgatacaga ctacaagagg tgatatccgc
     7741 caggttatta atctactttc aacgatatct acgactacta aaaccataaa ccatgaaaat
     7801 atcaacgaga tctcaaaggc atgggaaaag aatattgccc taaaaccctt tgacattgcc
     7861 cataaaatgt tggatggaca aatatactca gacataggtt caaggaactt tacactgaat
     7921 gataagatcg cattatattt tgatgacttt gatttcacac ccctaatgat tcaggaaaac
     7981 tatttatcta caagaccaag cgttttgaaa cccggccagt cacatttgga agcagttgct
     8041 gaggcggcta actgtatttc attaggtgat attgtcgaaa agaagattcg tagtagtgag
     8101 caattatgga gtcttttgcc tttgcatgct gttctttcat ctgtttatcc agcatcaaaa
     8161 gttgccggcc atatggcggg aagaatcaat ttcacagctt ggttgggcca aaattctaaa
     8221 tctgccaaat attataggtt actgcaagag atacattatc atacaagact aggtacatct
     8281 actgataaaa ttggactaag gctggattac ctaccaactt ttagaaaaag attattggac
     8341 ccatttttga agcaaggtgc tgatgcaatt tcatctgtca tagaggtaat ggacgattac
     8401 tatttgacca aagaagactg ggattctatt atggagtttt tcgtaggtcc tgatgtgacc
     8461 acagccatta tcaaaaagat accagctacg gttaaaagtg gattcacgcg gaaatacaac
     8521 agtatgacac atccagtcgc aatttacaga acaggtagta ctattggtgg tggtggtgtt
     8581 ggcaccagta ctagcacccc cgatttcgaa gatgtcgttg acgcagatga caatccagtt
     8641 cccgcagatg atgaagaaac acaagatagt agcacagact tgaagaagga taaacttatc
     8701 aagcagaaag ccaaacctac gaagaggaag actgccacca gtaaacctgg tggtagcaaa
     8761 aaaaggaaaa cgaaagcatg atttattcat acactaagat tataattaca cttttcttct
     8821 cattgatttt gtgagccgtt ctatcacaga tacgaaaact gtgtggtgat ccggctgcta
     8881 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     8941 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     9001 gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     9061 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     9121 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     9181 aacatgagaa ttcgagctcc agctt
//
