LOCUS       sequence-248939-        7547 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian linearizer gene circuit in a Flp-In expression system 
            based on negative feedback. The hTetR and EGFP is separated by a P2A
            motif..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7547)
  AUTHORS   Farquhar KS, Charlebois DA, Szenk M, Cohen J, Nevozhay D, Balazsi G
  TITLE     Role of network-mediated stochasticity in mammalian drug resistance.
  JOURNAL   Nat Commun. 2019 Jun 24;10(1):2766. doi: 10.1038/s41467-019-10330-w.
  PUBMED    31235692
REFERENCE   2  (bases 1 to 7547)
  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: "Nat 
            Commun."; date: "2019-06-24"; volume: "10(1):2766. doi"; pages: " 
            10.1038/s41467-019-10330-w"
COMMENT     Sequence Label: pDN-D2irTN2AG5kwh
FEATURES             Location/Qualifiers
     source          1..7547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    580..598
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    606..624
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          807..1379
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      1391..1400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2048..2104
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2105..2107,2108..2110,2111..2824)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2105..2107,2108..2110,2111..2821)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2150..2171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2411..2430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2758..2779
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2892..2912)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(3265..3282)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3271..3495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3628..3647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3779..3826
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3834..4856
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3834..4856
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4986..5107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5023..5042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5077..5096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5156..5172)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5156..5172)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5169..5191)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5180..5196
                     /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(5204..5210,5211..5228,5229..5234))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5249..5270
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5387..5404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5558..6146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5638..5657)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6317..7108,7109..7177))
                     /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     6940..6959
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7178..7282)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7357..7376)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcat ccctatcagt gatagagatc
      601 agatctccct atcagtgata gagagctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gactctagcg tttaaactta agcttggtac ccggggatcc tctagggcct
      721 ctgagctatt ccagaagtag tgaagaggct tttttggagg cctaggcttt tgcaaaaagc
      781 tccggatcga tcctgagaac ttcagggtga gtttggggac ccttgattgt tctttctttt
      841 tcgctattgt aaaattcatg ttatatggag ggggcaaagt tttcagggtg ttgtttagaa
      901 tgggaagatg tcccttgtat caccatggac cctcatgata attttgtttc tttcactttc
      961 tactctgttg acaaccattg tctcctctta ttttcttttc attttctgta actttttcgt
     1021 taaactttag cttgcatttg taacgaattt ttaaattcac ttttgtttat ttgtcagatt
     1081 gtaagtactt tctctaatca cttttttttc aaggcaatca gggtatatta tattgtactt
     1141 cagcacagtt ttagagaaca attgttataa ttaaatgata aggtagaata tttctgcata
     1201 taaattctgg ctggcgtgga aatattctta ttggtagaaa caactacatc ctggtcatca
     1261 tcctgccttt ctctttatgg ttacaatgat atacactgtt tgagatgagg ataaaatact
     1321 ctgagtccaa accgggcccc tctgctaacc atgttcatgc cttcttcttt ttcctacagg
     1381 tcctgcaggc gccaccatgg gttctagact ggacaagagc aaagtcataa actctgctct
     1441 ggaattactc aatgaagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct
     1501 gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc
     1561 cctggcaatc gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc
     1621 atggcaagac tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg
     1681 cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga
     1741 aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct
     1801 gtccgccgtg ggccacttta cactgggctg cgtattggag gatcaggagc atcaagtagc
     1861 aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat
     1921 tgagctgttc gaccatcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat
     1981 atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccaaaaaaga agagaaaggg
     2041 aagcggagct actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg
     2101 acctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2161 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2221 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2281 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2341 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2401 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2461 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2521 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2581 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2641 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2701 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2761 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2821 gtaaaccggt gcggccgcaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     2881 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     2941 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     3001 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     3061 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     3121 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     3181 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcctc gagtctagag
     3241 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3301 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3361 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3421 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3481 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     3541 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3601 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3661 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3721 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga
     3781 agttcctatt ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc
     3841 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     3901 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     3961 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     4021 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     4081 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     4141 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     4201 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     4261 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     4321 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     4381 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     4441 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     4501 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     4561 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     4621 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     4681 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     4741 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     4801 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg
     4861 tactacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4921 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4981 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5041 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5101 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5161 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5221 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5281 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5341 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5401 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5461 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5521 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5581 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5641 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5701 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5761 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5821 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5881 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5941 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6001 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6061 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6121 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6181 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6241 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6301 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6361 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6421 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6481 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6541 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6601 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6661 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6721 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6781 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6841 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6901 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6961 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7021 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7081 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7141 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7201 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7261 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7321 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     7381 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     7441 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     7501 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
