LOCUS       sequence-422254-        9145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of Osh1-GFP as an additional copy in genome, expressed 
            under CUP1 promoter. Uses auxotrophic marker URA3 (Saccharomyces 
            cerevisiae) for selection. Yeast PMN pathway substrate..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9145)
  AUTHORS   He CW, Cui XF, Ma SJ, Xu Q, Ran YP, Chen WZ, Mu JX, Li H, Zhu J, 
            Gong Q, Xie Z
  TITLE     Membrane recruitment of Atg8 by Hfl1 facilitates turnover of 
            vacuolar membrane proteins in yeast cells approaching stationary 
            phase.
  JOURNAL   BMC Biol. 2021 Jun 4;19(1):117. doi: 10.1186/s12915-021-01048-7.
  PUBMED    34088313
REFERENCE   2  (bases 1 to 9145)
  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: "BMC Biol.";
            date: "2021-06-4"; volume: "19(1):117. doi"; pages: " 
            10.1186/s12915-021-01048-7"
COMMENT     Sequence Label: PCUP1-GFP-Osh1-406
FEATURES             Location/Qualifiers
     source          1..9145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..51
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(89..107)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        175..279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(280..348,349..1140)
                     /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(498..517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1311..1899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1800..1819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2053..2070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2187..2208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2223..2228,2229..2246,2247..2253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2261..2277
                     /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     2266..2288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2285..2301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2285..2301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2320..2340
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2322..2340
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2359..2593
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             2604..3317
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     primer_bind     complement(7240..7259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7241..7259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7269..7286)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7269..7285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7278..7300)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7426..7881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7513..7532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7723..7744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(8012..8815)
                     /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(8816..9031)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(9055..9074)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
       61 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
      121 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      241 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      301 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      361 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      421 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      481 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
      541 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      601 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      661 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      721 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      781 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
      841 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      901 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      961 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1021 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1081 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1141 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1201 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1261 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1441 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1681 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1921 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1981 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2041 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2101 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2161 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     2221 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     2281 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     2341 gaacaaaagc tggagctcct agttagaaaa agacattttt gctgtcagtc actgtcaaga
     2401 gattcttttg ctggcatttc ttctagaagc aaaaagagcg atgcgtcttt tccgctgaac
     2461 cgttccagca aaaaagacta ccaacgcaat atggattgtc agaatcatat aaaagagaag
     2521 caaataactc cttgtcttgt atcaattgca ttataatatc ttcttgttag tgcaatatca
     2581 tatagaagtc atcgactagt aacatgtcta aaggtgaaga attattcact ggtgttgtcc
     2641 caattttggt tgaattagat ggtgatgtta atggtcacaa attttctgtc tccggtgaag
     2701 gtgaaggtga tgctacttac ggtaaattga ccttaaaatt tatttgtact actggtaaat
     2761 tgccagttcc atggccaacc ttagtcacta ctttaactta tggtgttcaa tgtttttcta
     2821 gatacccaga tcatatgaaa caacatgact ttttcaagtc tgccatgcca gaaggttatg
     2881 ttcaagaaag aactattttt ttcaaagatg acggtaacta caagaccaga gctgaagtca
     2941 agtttgaagg tgatacctta gttaatagaa tcgaattaaa aggtattgat tttaaagaag
     3001 atggtaacat tttaggtcac aaattggaat acaactataa ctctcacaat gtttacatca
     3061 tggctgacaa acaaaagaat ggtatcaaag ttaacttcaa aattagacac aacattgaag
     3121 atggttctgt tcaattagct gaccattatc aacaaaatac tccaattggt gatggtccag
     3181 tcttgttacc agacaaccat tacttatcca ctcaatctgc cttatccaaa gatccaaacg
     3241 aaaagagaga ccacatggtc ttgttagaat ttgttactgc tgctggtatt acccatggta
     3301 ttgatgaatt gtacaaagga gccggaacaa gtgacatgga acaacctgat ctatcgtctg
     3361 tggccatcag taagccgctg ctgaagttga aacttctcga cgcccttcgc cagggaagtt
     3421 tccccaacct acaagatctc ctaaagaaac aattccagcc gctagacgac ccaaacgtcc
     3481 aacaagtgct ccatctcatg ctccactatg ccgtgcaagt cgcccccatg gctgtcataa
     3541 aggaaatcgt ccatcattgg gtctcaacta caaacaccac ttttctaaac atccatcttg
     3601 atctaaacga acgggactcc aacggcaaca ccccattgca catcgccgcc taccagtccc
     3661 gcggtgatat cgtagccttc ctcctggacc aaccaaccat caacgactgc gtgctcaaca
     3721 actcccactt gcaggccatc gaaatgtgca agaacctaaa catcgcgcag atgatgcagg
     3781 tgaaacgctc cacatacgtt gcagagaccg cccaggaatt cagaacagct tttaacaaca
     3841 gggacttcgg ccacctagaa tctatcctct ccagccctcg aaacgcagaa ctgctcgaca
     3901 tcaacggtat ggacccggag actggcgata ccgttctgca cgaattcgtc aagaaaagag
     3961 acgtcatcat gtgccgttgg ttgcttgaac acggtgctga ccccttcaag agagaccgca
     4021 agggcaaact gcccatcgag ctcgttagga aagtcaatga aaacgacacc gccaccaaca
     4081 ccaagatcgc catcgacatc gaactgaaaa aactattgga aagggccacc agggagcaaa
     4141 gtgtcatcga cgtcacaaac aacaacttgc acgaggcccc cacttacaaa ggctacctga
     4201 aaaaatggac caacttcgct caaggctaca aattgcgttg gttcatcctt agtagcgatg
     4261 ggaaactatc ctactacatc gatcaggccg acactaagaa tgcctgcagg ggctccctaa
     4321 acatgtcttc gtgctctctg catttggatt cgtctgaaaa gttgaaattc gaaattatcg
     4381 gcggtaacaa cggtgttatc aggtggcatt taaaggggaa ccaccccatc gagacaaata
     4441 gatgggtttg ggccatccag ggcgccataa gatacgcaaa ggacagagaa attttgctgc
     4501 acaatggccc ctattcgcct tctctggcct taagccatgg cttgtcatcc aaagtgtcca
     4561 ataaagaaaa cttgcatgca acttcaaaac ggttgaccaa gagcccgcat ctgtccaaat
     4621 ccacactgac acaaaacgat cacgataatg acgatgacag cactaacaac aacaacaaca
     4681 aaagtaataa tgattatgac gataataata ataataataa taatgacgat gatgattatg
     4741 atgatgatga tgaaagtaga cccctcatag aaccattacc gttgatttca tccagaagcc
     4801 aaagcttaag cgaaatcact cccggtccac attctaggaa gtctacagtc tcgtctacaa
     4861 gggcagccga tataccatca gatgatgagg gttactctga ggacgattct gatgacgacg
     4921 gtaactcctc ttacacaatg gaaaacggcg gtgaaaacga tggcgacgaa gatctaaatg
     4981 ccatttatgg tccctatatt caaaaactac acatgctaca aagatccatt tccatcgagt
     5041 tggcatcttt gaacgaattg ctgcaagata aacaacaaca cgatgagtac tggaacaccg
     5101 tcaacacttc tattgaaacc gtcagcgaat ttttcgacaa attaaatcgg ttgacctctc
     5161 aaagggaaaa aagaatgatt gcccaaatga ccaagcaacg ggatgttaac aatgtttgga
     5221 ttcaatcggt aaaagatctg gaaatggaac tggttgataa agacgaaaaa ttggttgcct
     5281 tggataaaga acggaaaaat ctgaaaaaaa tgcttcaaaa aaaattgaac aatcaaccac
     5341 aggttgaaac tgaggctaat gaagaatccg atgatgcaaa ttcaatgata aaaggatccc
     5401 aagaatcaac aaataccctt gaggaaatcg taaaatttat cgaagcaaca aaggaaagtg
     5461 atgaggattc tgacgccgac gaatttttcg acgcagaaga agctgcttcc gacaaaaaag
     5521 ccaatgattc ggaagactta accacaaaca aggagactcc agctaatgcg aaaccacaag
     5581 aagaagctcc tgaagacgag agccttattg tgatcagttc tccacaggtg gaaaagaaga
     5641 accaactatt aaaagaggga tcattcgtcg gatatgaaga cccagtgaga accaaactgg
     5701 ctttagacga agataatcgt cccaagattg gtctctggtc tgttttaaag tctatggtcg
     5761 gtcaagactt aaccaaacta actctaccgg tatcgttcaa tgagccaaca tccttactac
     5821 agagagtatc tgaagatatt gagtattctc atattcttga ccaagctgcc acttttgaag
     5881 actcctcttt aagaatgcta tatgtagctg cctttactgc atcaatgtac gcatctacca
     5941 ctaacagagt gtctaaacca ttcaacccct tactcggtga aacttttgaa tatgccagaa
     6001 ctgatggtca gtaccgattc ttcaccgaac aagtctctca ccacccacct atctctgcta
     6061 cttggacaga atcgcccaaa tgggattttt acggtgaatg taatgttgat tcgtcattca
     6121 atgggcgcac gttcgccgtg caacatttag gattatggta cattactatc cggcctgatc
     6181 ataatattag tgttcccgag gaaacttatt cctggaaaaa accaaataac actgttatcg
     6241 gtattttaat ggggaaacca caagtagaca acagtgggga cgtcaaagtc acaaaccata
     6301 ccacaggcga ctattgtatg ctgcattaca aagcccatgg ctggacctca gccggtgcat
     6361 atgaagtcag aggtgaagta ttcaacaagg acgataaaaa attatgggtt cttggtgggc
     6421 attggaatga ttccatttac gggaaaaaag taactgctag aggcggagaa ctgacattag
     6481 acagaataaa aacggcaaat tctgccacgg gaggaccaaa actagatggg tctaagtttc
     6541 tgatatggaa agcaaatgaa aggccttcag tgccatttaa tttaacgtcg tttgcattga
     6601 ctttgaatgc tttgccaccc cacttgatac catatttagc acccacagat agtcgtttaa
     6661 ggcccgatca aagggctatg gaaaatggtg aatacgataa agctgccgcg gaaaagcatc
     6721 gtgttgaagt aaaacaaagg gcagcaaaaa aagaaaggga acaaaaagga gaagaataca
     6781 gacctaagtg gtttgtccag gaggagcacc ccgttaccaa aagtctatac tggaaattta
     6841 atggagagta ttggaacaaa agaaaaaatc atgactttaa agattgtgct gatattttct
     6901 aagctgtgca atgtagtcac aataacactc gttcatttgt atccattgcg aatgccggta
     6961 catcggaaaa caggatagga cctatttaat tatatagtat gaagtattca taacttcttg
     7021 aggcatcaat acatcatatt ccatgagctg cgtggcattc atactcattg atttaaaggt
     7081 tttttatttt catggaaaag attaaccggg ctgaacgaaa tatattaaag atgctaaaac
     7141 ttatgctttc attgactttc aatagtgtcc actaaccaaa aaaaaaaact actctaacaa
     7201 gggatcccca tggattcaaa gccgatggta cccaattcgc cctatagtga gtcgtattac
     7261 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     7321 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     7381 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg
     7441 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     7501 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     7561 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     7621 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     7681 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     7741 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     7801 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     7861 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     7921 ttcacaccgc atagggtaat aactgatata attaaattga agctctaatt tgtgagttta
     7981 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     8041 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     8101 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     8161 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     8221 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     8281 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     8341 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     8401 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     8461 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     8521 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     8581 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     8641 ttagtaaaca aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac
     8701 gaacatccaa tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg
     8761 cagcaacagg actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt
     8821 atcttcgttt cctgcaggtt tttgttctgt gcagttgggt taagaatact gggcaatttc
     8881 atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc
     8941 gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa
     9001 aaaaagaata aaaaaaaaat gatgaattga aaaggtggta tggtgcactc tcagtacaat
     9061 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     9121 ctgacgggct tgtctgctcc cggca
//
