LOCUS       sequence-349804-        8323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA targeting hNKD2 - encodes puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8323)
  AUTHORS   Wu B, Woo JS, Vila P, Jew M, Leung J, Sun Z, Srikanth S, Gwack Y
  TITLE     NKD2 mediates stimulation-dependent ORAI1 trafficking to augment 
            Ca(2+) entry in T cells.
  JOURNAL   Cell Rep. 2021 Aug 24;36(8):109603. doi: 
            10.1016/j.celrep.2021.109603.
  PUBMED    34433025
REFERENCE   2  (bases 1 to 8323)
  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: "Cell Rep.";
            date: "2021-08-24"; volume: "36(8):109603. doi"; pages: " 
            10.1016/j.celrep.2021.109603"
COMMENT     Sequence Label: pLentiguide-puro_hNKD2sg#1
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     381..398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    515..536
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(551..556,557..574,575..581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    589..605
                     /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     594..616
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     613..629
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     648..668
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        650..668
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        696..922
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             923..1103
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1150..1275
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1768..2001
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2186..2230
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        2408..2656
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2408..2428
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2579..2598
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2678..2753
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2809..2926
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3065..4243
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3296..4234
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4191..4211
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4256..4855
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4256..4275)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4752..4772
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4871..5459
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4924..4944)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5342..5353)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5531..5764
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5842..5963
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5879..5898)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5933..5952
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6003..6138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6158..6177)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6159..6177)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6187..6204)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6187..6203)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6196..6218)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6345..6800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6432..6451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6642..6663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6826..6930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6931..6999,7000..7791)
                     /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(7149..7168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7962..8323,1..227)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
       61 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
      121 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      181 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      241 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
      301 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
      361 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
      421 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
      481 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      541 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      601 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      661 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
      721 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
      781 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
      841 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
      901 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
      961 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     1021 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     1081 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     1141 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     1201 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     1261 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     1321 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     1381 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     1441 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     1501 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     1561 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     1621 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     1681 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     1741 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     1801 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     1861 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     1921 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     1981 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     2041 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     2101 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     2161 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     2221 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     2281 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     2341 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     2401 acccagagag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
     2461 tagagagata attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
     2521 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
     2581 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
     2641 gaaaggacga aacaccgccg ggctctctct ccgcttggtt ttagagctag aaatagcaag
     2701 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttta
     2761 agcttggcgt aactagatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2821 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2881 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2941 ggacagcaga gatccacttt ggcgccggct cgagggggcc cgggtgcaaa gatggataaa
     3001 gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg
     3061 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     3121 ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga
     3181 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     3241 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     3301 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     3361 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
     3421 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
     3481 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
     3541 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
     3601 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
     3661 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
     3721 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
     3781 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
     3841 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
     3901 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     3961 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     4021 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     4081 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     4141 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     4201 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgac
     4261 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     4321 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     4381 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     4441 caaggtgtgg gtcgcggacg acggcgccgc cgtggcggtc tggaccacgc cggagagcgt
     4501 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     4561 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     4621 gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     4681 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     4741 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     4801 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg
     4861 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     4921 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     4981 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5041 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5101 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5161 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5221 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     5281 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5341 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5401 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     5461 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5521 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     5581 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5641 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5701 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5761 agcagtacgt atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa
     5821 tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc
     5881 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5941 tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc
     6001 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6061 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     6121 gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc
     6181 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     6241 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     6301 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     6361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6781 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6841 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6901 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6961 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     7021 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     7081 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     7141 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     7201 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     7261 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     7321 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     7381 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     7441 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     7501 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     7561 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     7621 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     7681 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7741 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7801 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7861 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7921 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7981 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     8041 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     8101 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     8161 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     8221 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     8281 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cct
//
