LOCUS       sequence-259134-        8012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All in one overexpression of CasRx and guideRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8012)
  TITLE     Unpublished
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8012)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAK_DR30_CasRx_GFP
FEATURES             Location/Qualifiers
     source          1..8012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..268
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     20..40
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     191..210
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        270..299
                     /label=RfxCas13d DR30
                     /note="5' processed 30-base direct repeat for an RfxCas13d 
                     gRNA (Konermann et al., 2018)"
     enhancer        448..733
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        735..1012
                     /label=chicken beta-actin promoter
     intron          1013..1240
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1264..1284
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1294..4191
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             4201..4221
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4231..4257
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4273..4326
                     /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"
     CDS             join(4327..4329,4330..5043)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4327..4329,4330..5040)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4369..4390)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4630..4649)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4977..4998
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5072..5089)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5078..5285
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5294..5434
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5294..5423
                     /label=AAV2 ITR
     rep_origin      5509..5964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5596..5615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5806..5827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5981..6000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6100..6122
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6160..6178)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6246..6350
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6351..6419,6420..7211)
                     /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(6569..6588)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7382..7970
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7871..7890
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtg agggcctatt tcccatgatt ccttcatatt tgcatatacg
       61 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
      121 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
      181 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
      241 atatatcttg tggaaaggac gaaacaccga acccctacca actggtcggg gtttgaaacg
      301 ggtcttcgag aagacctttt ttttggacag cagagatcca gtttggttaa ttaatgcaaa
      361 gatggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga
      421 aaggagtggg aattctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg
      481 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
      541 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      601 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
      661 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      721 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
      781 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
      841 gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc
      901 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      961 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
     1021 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
     1081 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     1141 taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg tattaatgtt
     1201 taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg tgccaccatg
     1261 agccccaaga agaagagaaa ggtggaggcc agcatcgaaa aaaaaaagtc cttcgccaag
     1321 ggcatgggcg tgaagtccac actcgtgtcc ggctccaaag tgtacatgac aaccttcgcc
     1381 gaaggcagcg acgccaggct ggaaaagatc gtggagggcg acagcatcag gagcgtgaat
     1441 gagggcgagg ccttcagcgc tgaaatggcc gataaaaacg ccggctataa gatcggcaac
     1501 gccaaattca gccatcctaa gggctacgcc gtggtggcta acaaccctct gtatacagga
     1561 cccgtccagc aggatatgct cggcctgaag gaaactctgg aaaagaggta cttcggcgag
     1621 agcgctgatg gcaatgacaa tatttgtatc caggtgatcc ataacatcct ggacattgaa
     1681 aaaatcctcg ccgaatacat taccaacgcc gcctacgccg tcaacaatat ctccggcctg
     1741 gataaggaca ttattggatt cggcaagttc tccacagtgt atacctacga cgaattcaaa
     1801 gaccccgagc accatagggc cgctttcaac aataacgata agctcatcaa cgccatcaag
     1861 gcccagtatg acgagttcga caacttcctc gataacccca gactcggcta tttcggccag
     1921 gcctttttca gcaaggaggg cagaaattac atcatcaatt acggcaacga atgctatgac
     1981 attctggccc tcctgagcgg actgaggcac tgggtggtcc ataacaacga agaagagtcc
     2041 aggatctcca ggacctggct ctacaacctc gataagaacc tcgacaacga atacatctcc
     2101 accctcaact acctctacga caggatcacc aatgagctga ccaactcctt ctccaagaac
     2161 tccgccgcca acgtgaacta tattgccgaa actctgggaa tcaaccctgc cgaattcgcc
     2221 gaacaatatt tcagattcag cattatgaaa gagcagaaaa acctcggatt caatatcacc
     2281 aagctcaggg aagtgatgct ggacaggaag gatatgtccg agatcaggaa aaatcataag
     2341 gtgttcgact ccatcaggac caaggtctac accatgatgg actttgtgat ttataggtat
     2401 tacatcgaag aggatgccaa ggtggctgcc gccaataagt ccctccccga taatgagaag
     2461 tccctgagcg agaaggatat ctttgtgatt aacctgaggg gctccttcaa cgacgaccag
     2521 aaggatgccc tctactacga tgaagctaat agaatttgga gaaagctcga aaatatcatg
     2581 cacaacatca aggaatttag gggaaacaag acaagagagt ataagaagaa ggacgcccct
     2641 agactgccca gaatcctgcc cgctggccgt gatgtttccg ccttcagcaa actcatgtat
     2701 gccctgacca tgttcctgga tggcaaggag atcaacgacc tcctgaccac cctgattaat
     2761 aaattcgata acatccagag cttcctgaag gtgatgcctc tcatcggagt caacgctaag
     2821 ttcgtggagg aatacgcctt tttcaaagac tccgccaaga tcgccgatga gctgaggctg
     2881 atcaagtcct tcgctagaat gggagaacct attgccgatg ccaggagggc catgtatatc
     2941 gacgccatcc gtattttagg aaccaacctg tcctatgatg agctcaaggc cctcgccgac
     3001 accttttccc tggacgagaa cggaaacaag ctcaagaaag gcaagcacgg catgagaaat
     3061 ttcattatta ataacgtgat cagcaataaa aggttccact acctgatcag atacggtgat
     3121 cctgcccacc tccatgagat cgccaaaaac gaggccgtgg tgaagttcgt gctcggcagg
     3181 atcgctgaca tccagaaaaa acagggccag aacggcaaga accagatcga caggtactac
     3241 gaaacttgta tcggaaagga taagggcaag agcgtgagcg aaaaggtgga cgctctcaca
     3301 aagatcatca ccggaatgaa ctacgaccaa ttcgacaaga aaaggagcgt cattgaggac
     3361 accggcaggg aaaacgccga gagggagaag tttaaaaaga tcatcagcct gtacctcacc
     3421 gtgatctacc acatcctcaa gaatattgtc aatatcaacg ccaggtacgt catcggattc
     3481 cattgcgtcg agcgtgatgc tcaactgtac aaggagaaag gctacgacat caatctcaag
     3541 aaactggaag agaagggatt cagctccgtc accaagctct gcgctggcat tgatgaaact
     3601 gcccccgata agagaaagga cgtggaaaag gagatggctg aaagagccaa ggagagcatt
     3661 gacagcctcg agagcgccaa ccccaagctg tatgccaatt acatcaaata cagcgacgag
     3721 aagaaagccg aggagttcac caggcagatt aacagggaga aggccaaaac cgccctgaac
     3781 gcctacctga ggaacaccaa gtggaatgtg atcatcaggg aggacctcct gagaattgac
     3841 aacaagacat gtaccctgtt cagaaacaag gccgtccacc tggaagtggc caggtatgtc
     3901 cacgcctata tcaacgacat tgccgaggtc aattcctact tccaactgta ccattacatc
     3961 atgcagagaa ttatcatgaa tgagaggtac gagaaaagca gcggaaaggt gtccgagtac
     4021 ttcgacgctg tgaatgacga gaagaagtac aacgataggc tcctgaaact gctgtgtgtg
     4081 cctttcggct actgtatccc caggtttaag aacctgagca tcgaggccct gttcgatagg
     4141 aacgaggccg ccaagttcga caaggagaaa aagaaggtgt ccggcaattc cggatccgga
     4201 cctaagaaaa agaggaaggt ggcggccgct tacccatacg atgttccaga ttacgctgct
     4261 agcggcagtg gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct
     4321 ggcccagtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     4381 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     4441 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     4501 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     4561 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     4621 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     4681 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     4741 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     4801 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     4861 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     4921 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     4981 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     5041 taacgtacgc taactagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     5101 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5161 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5221 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     5281 ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5341 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     5401 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     5461 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     5521 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5581 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5641 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5701 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5761 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5821 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     5881 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5941 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     6001 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6061 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6121 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6181 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6241 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6301 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6361 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6421 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6481 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6541 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6601 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6661 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6721 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6781 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6841 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6901 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6961 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7021 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     7081 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7141 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7201 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7261 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7321 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7381 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7441 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7501 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7561 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7621 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7681 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7741 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7801 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7861 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7921 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7981 acgcggcctt tttacggttc ctggcctttt gc
//
