LOCUS       sequence-346992-        8720 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Opto-kinesin for optogenetic heterodimerization to iLID via 
            SSPB(micro).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8720)
  AUTHORS   Nijenhuis W, van Grinsven MMP, Kapitein LC
  TITLE     An optimized toolbox for the optogenetic control of intracellular 
            transport.
  JOURNAL   J Cell Biol. 2020 Apr 6;219(4). pii: 133834. doi: 
            10.1083/jcb.201907149.
  PUBMED    32328628
REFERENCE   2  (bases 1 to 8720)
  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: "J Cell 
            Biol."; date: "2020-04-6"; volume: "219(4). pii: 133834. doi"; 
            pages: " 10.1083/jcb.201907149"
COMMENT     Sequence Label: pB80-KIF1A(1-365)-VVDfast-mVenus-SSPB(micro)
FEATURES             Location/Qualifiers
     source          1..8720
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1045..1067
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1130..1139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2112..2121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2735..2737,2738..3448)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2735..2737,2738..3448)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2777..2798)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3038..3057)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3385..3406
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(3578..3784,3785..3787,3788..3907)
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein and with the mutation R74Q to reduce the 
                     affinity for iLID"
                     /label=SspB micro
                     /note="dimerizes with iLID at moderate affinity to create 
                     iLID micro (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNAQFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     polyA_signal    4040..4174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4077..4096)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4131..4150
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4188..4204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4188..4204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4201..4223)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4212..4228
                     /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(4236..4242,4243..4260,4261..4266))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4281..4302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4419..4436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4590..5178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4670..4689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5349..6140,6141..6209))
                     /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     5972..5991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6210..6314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6382..6400
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6438..6460)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6560..6579
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6773..6795
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6787..6804
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6788..6804
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
       61 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      121 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      181 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      241 ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa agggctccgg
      301 gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
      361 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
      421 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
      481 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
      541 agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct ccccgagttg
      601 ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg
      661 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
      721 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
      781 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
      841 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
      901 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
      961 cgcgccgccg tccccttctc catctccagc ctcggggctg tccgcagggg gacggctgcc
     1021 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggggtttata
     1081 tcttcccttc tctgttcctc cgcagccccc aagctaggca attgggcgcg ccaccatggc
     1141 cggggcttcg gtgaaggtgg cggtgcgtgt ccgccccttc aactcccggg aaatgagccg
     1201 agactccaag tgcatcattc agatgtctgg aagcaccacc actattgtca accccaaaca
     1261 gcccaaggag acacccaaga gcttcagctt tgattactcc tactggtcac atacctcgcc
     1321 ggaggacatc aactatgcct ctcagaagca ggtctacagg gacattgggg aagagatgct
     1381 acagcatgcc tttgagggct acaatgtgtg catctttgcc tatgggcaga cgggggccgg
     1441 caagtcctac accatgatgg ggaaacagga gaaggaccag caaggcatca ttccacagct
     1501 ctgcgaagac ctcttctctc gaatcaacga caccaccaac gacaacatgt cctactctgt
     1561 ggaggtcagc tacatggaga tttactgtga gcgagtccgt gacctcctga accccaagaa
     1621 caaaggcaac ctgcgtgtga gggagcaccc tctgctgggg ccttatgtgg aggacctctc
     1681 caagctggct gtaacctcct acaatgacat ccaggacctc atggactcag ggaacaaggc
     1741 caggactgtg gcggccacaa acatgaatga gaccagcagc cgttctcacg cagtcttcaa
     1801 catcatcttt acccagaagc gccatgatgc agagaccaac atcaccacag agaaagtgag
     1861 caaaatcagc ctggtggacc tggcaggcag tgaacgagcc gactccacag gagccaaggg
     1921 cacgcgactc aaggaggggg ccaacatcaa caagtctttg accaccctgg gaaaggtcat
     1981 ctctgctctg gctgaaatgg actctggacc caacaagaac aagaaaaaga agaagacaga
     2041 tttcatccct taccgagact ccgtattgac atggcttctc cgggaaaacc tcggtggcaa
     2101 ctcaaggaca gccatggtgg cagccctgag ccctgcggac atcaactatg atgagaccct
     2161 cagcacactg agatatgcag atcgggccaa acaaatccgc tgcaatgcca tcatcaatga
     2221 agaccccaac tctagaggtg gcgggggcag tgggggatcc catactcttt atgcccccgg
     2281 tggatatgac attatgggat atctgtgcca gatcatgaac cggccaaacc cgcaggtgga
     2341 actgggcccc gtggatacat cctgcgcctt ggttctttgt gacctgaaac agaaagacac
     2401 cccggtagtt tacgcgagtg aagccttcct ctacatgaca ggttacagca acgcagaggt
     2461 gctgggccgg aattgccggt ttctgcaaag ccctgacggc atggtgaagc ccaagagcac
     2521 ccggaagtac gtggatagta acacaatcaa tactatgcgc aaggcaatcg acaggaatgc
     2581 cgaggtgcag gttgaagtag tcaattttaa aaagaatgga cagcgatttg ttaatttcct
     2641 gactatgata cctgttaggg acgaaacagg cgagtatcga tactctatgg gattccagtg
     2701 cgaagtcgac gggagcgggg gtgggagtaa gcttgtgagc aagggcgagg agctgttcac
     2761 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2821 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tgatctgcac
     2881 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctcggct acggcctgca
     2941 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3001 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3061 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3121 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3181 cgtctatatc atggccgaca agcagaagaa cggcatcaag gcaaacttca agatccgcca
     3241 caacatcgag gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3301 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca aactgagcaa
     3361 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3421 cactctcggc atggacgagc tgtacaaggc tagcggaggc ggtggctcag gcggaggagg
     3481 gtcaggaggg ggaggcagtg gcgggggcgg ctctggagga gggggttctg gtggtggtag
     3541 ctacgtagat ctgagcggcc tgaccctgca ggaattcagc tccccgaaac gccctaagct
     3601 gctgcgtgaa tattacgatt ggctggttga taacagcttt accccatatc tggtggtgga
     3661 tgccacatac ctgggcgtga acgtgcccgt ggagtatgtg aaagacggtc agatcgtgct
     3721 gaatctgtct gcaagtgcga ccggcaacct gcaactgaca aatgatttta tccagttcaa
     3781 cgcccagttt aagggcgtgt ctcgtgaact gtatatcccg atgggtgccg ctctggccat
     3841 ttacgctcgc gagaacggcg atggtgtgat gttcgaacca gaagaaatct atgacgagct
     3901 gaatattggt taaaccggtc ctagataagt aatgatcata atcagccata tcacatctgt
     3961 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     4021 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4081 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4141 caaactcatc aatgtatctt atcatgtctg gatcaattcg taatcatgtc atagctgttt
     4201 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4261 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4321 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4381 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5221 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5281 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5341 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5401 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5461 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5521 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5581 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5641 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5701 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5761 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5821 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5881 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5941 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6001 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6061 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6121 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6181 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6241 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6301 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6361 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6421 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6481 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6541 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     6601 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     6661 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6721 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6781 cgacgttgta aaacgacggc cagtgccaag ttgggatctt tgcattggcc cacggctctc
     6841 aggatgggga tgctcccctt cagcacccgg ttccccttgg aaactgatgg tcctggctct
     6901 gtggcatggc agtggcactg tgaggagccc cctaccagca gcacacagtg gggttggcac
     6961 tgccacgctc cggatgccgc gctctgatcc aaccccataa tcaagggaac ccgaattgcc
     7021 ccatcattgc ccccaccacc cccatcctgc cgggccctca caccccacgc tgccttctgg
     7081 tgacattccc cagcccaaac ccacggcttc atggctaccg cggggcattt cccattgccg
     7141 ccccattatc agctctgcac acctcccgct gtacccatgc ctcgtggctg cccttctttg
     7201 acgtataatc ttctaattaa tacccggcct tgtcaaagtg gagcacaaac gttaattaat
     7261 tccccagcag gcaggtaatt aacagtgtga ctcccttttt gctgcgagtg gggctgatac
     7321 agagagatgt ggcactatgg agcccacggg gtcctggcac tgggtgccca cggaggtccc
     7381 catgtgctgc agtgtcaccg cctccgagga gacagtattg tccctgcggt gtccctgcag
     7441 ctcagctctg tccacagggc cacctccagt ttggagggga cacaatgcag ccccgatgca
     7501 acccatcctc gcagcatccc agggacaaag accccactgc aagaccgcac acagggctgg
     7561 gtcccgctcc cctaatatct acagtgcttc ttgcatggcc ccttaatcaa tgcagttaat
     7621 cagcatgcgc tcatgcaccg ctctggagct gcaaagcccc tcgcagcgct gctcaccaac
     7681 accgcgcacc gccccggccc agcctgcagc acgcgctgca aacaggaaag aaacaaaata
     7741 ttgcccaaat gtaggcaaag gcattcggct gccttgacct ccgccgggcc gggccctgcc
     7801 tgactcagct ccttactcag cgctcgcttc ctccctccgg ctgccaccgc cgcagcgcac
     7861 accctgacaa agagtggccc ttaacgggct ctgaggtgca cccagcagtg cactcagcag
     7921 tccaagggcc ggcctggagg tttgcaccgc tacgtgctga cattagcatt gaacttggcc
     7981 ctgggtagtg ctgcaggccg ggcggggtgg gtgtagagag tgcagcgcgc gttgcacccg
     8041 gtgccccttc ccctcccttg catcccagca ggctgcaccc cagcaccagg cccgtgcatg
     8101 catgctcctg gtgttattgc agcctggtgc atgcatgcgt cttgatggtg cagcgctgtg
     8161 catgcatcct ccttggtgtg tagcagtcta gtgcatgcat acccctcggt gttattgctg
     8221 ctctgtgcac gcacgctcat tgtatcactt catcccagtg catgcactca cactggagcg
     8281 attgctgctc ggtgcacgca cactcattgt atcactgcag ctcagtggct gcacgcacac
     8341 cggtgttatt gctgctcggt gcgtgcatgc acatcagtgt cgctgcagct cagtgcatgc
     8401 acgctcattg cccatcgcta tccctgcctc tcctgctggc gctccccggg aggtgacttc
     8461 aaggggaccg caggaccacc tcgggggtgg ggggagggct gcacacgcgg accccgctcc
     8521 ccctccccaa caaagcactg tggaatcaaa aaggggggag gggggatgga ggggcgcgtc
     8581 acacccccgc cccacaccct cacctcgagg tgagccccac gttctgcttc actctcccca
     8641 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     8701 cgatgggggc gggggggggg
//
