pCMV-hA1-BE3

LOCUS       pCMV-hA1-BE3              8553 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses hA1-BE3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-hA1-BE3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8553)
  AUTHORS   Wang X, Li J, Wang Y, Yang V, Wei J, Wu J, Wang R, Huang X, Chen J, 
            Yang L
  TITLE     Efficient base editing in methylated regions with a human 
            APOBEC3A-Cas9 fusion.
  JOURNAL   Nat Biotechnol.,  2018
REFERENCE   2  (bases 1 to 8553)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8553
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        134..513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        514..717
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     668..688
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     759..778
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        759..777
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1547..5647
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5660..5908
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5921..5941
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5950..5967
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5990..6007)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5996..6220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6291..6307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6291..6307)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6304..6326)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6315..6331
                     /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(6339..6369)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6384..6405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6522..6539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6693..7281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6773..6792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7452..8312)
                     /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     8075..8094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8313..8417)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8496..8515)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
       61 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
      121 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
      181 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      241 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      301 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      361 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      421 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      481 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      541 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      601 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      661 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
      721 ttagtgaacc gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag
      781 agccgccacc atgacttctg agaaaggtcc ttcaaccggt gaccccactc tgaggagaag
      841 aatcgaaccc tgggagtttg acgtcttcta tgaccccaga gaacttcgta aagaggcctg
      901 tctgctctac gaaatcaagt ggggcatgag ccggaagatc tggcgaagct caggcaaaaa
      961 caccaccaat cacgtggaag ttaattttat aaaaaaattt acgtcagaaa gagattttca
     1021 cccatccatg agctgctcca tcacctggtt cttgtcctgg agtccctgct gggaatgctc
     1081 ccaggctatt agagagtttc tgagtcggca ccctggtgtg actctagtga tctacgtagc
     1141 tcggcttttt tggcacatgg atcaacaaaa tcggcaaggt ctcagggacc ttgttaacag
     1201 tggagtaact attcagatta tgagagcatc agagtattat cactgctgga ggaattttgt
     1261 caactaccca cctggggatg aagctcactg gccacaatac ccacctctgt ggatgatgtt
     1321 gtacgcactg gagctgcact gcataattct aagtcttcca ccctgtttaa agatttcaag
     1381 aagatggcaa aatcatctta catttttcag acttcatctt caaaactgcc attaccaaac
     1441 gattccgcca cacatccttt tagctacagg gctgatacat ccttctgtgg cttggagaag
     1501 cggcagcgag actcccggga cctcagagtc cgccacaccc gaaagtgata aaaagtattc
     1561 tattggttta gccatcggca ctaattccgt tggatgggct gtcataaccg atgaatacaa
     1621 agtaccttca aagaaattta aggtgttggg gaacacagac cgtcattcga ttaaaaagaa
     1681 tcttatcggt gccctcctat tcgatagtgg cgaaacggca gaggcgactc gcctgaaacg
     1741 aaccgctcgg agaaggtata cacgtcgcaa gaaccgaata tgttacttac aagaaatttt
     1801 tagcaatgag atggccaaag ttgacgattc tttctttcac cgtttggaag agtccttcct
     1861 tgtcgaagag gacaagaaac atgaacggca ccccatcttt ggaaacatag tagatgaggt
     1921 ggcatatcat gaaaagtacc caacgattta tcacctcaga aaaaagctag ttgactcaac
     1981 tgataaagcg gacctgaggt taatctactt ggctcttgcc catatgataa agttccgtgg
     2041 gcactttctc attgagggtg atctaaatcc ggacaactcg gatgtcgaca aactgttcat
     2101 ccagttagta caaacctata atcagttgtt tgaagagaac cctataaatg caagtggcgt
     2161 ggatgcgaag gctattctta gcgcccgcct ctctaaatcc cgacggctag aaaacctgat
     2221 cgcacaatta cccggagaga agaaaaatgg gttgttcggt aaccttatag cgctctcact
     2281 aggcctgaca ccaaatttta agtcgaactt cgacttagct gaagatgcca aattgcagct
     2341 tagtaaggac acgtacgatg acgatctcga caatctactg gcacaaattg gagatcagta
     2401 tgcggactta tttttggctg ccaaaaacct tagcgatgca atcctcctat ctgacatact
     2461 gagagttaat actgagatta ccaaggcgcc gttatccgct tcaatgatca aaaggtacga
     2521 tgaacatcac caagacttga cacttctcaa ggccctagtc cgtcagcaac tgcctgagaa
     2581 atataaggaa atattctttg atcagtcgaa aaacgggtac gcaggttata ttgacggcgg
     2641 agcgagtcaa gaggaattct acaagtttat caaacccata ttagagaaga tggatgggac
     2701 ggaagagttg cttgtaaaac tcaatcgcga agatctactg cgaaagcagc ggactttcga
     2761 caacggtagc attccacatc aaatccactt aggcgaattg catgctatac ttagaaggca
     2821 ggaggatttt tatccgttcc tcaaagacaa tcgtgaaaag attgagaaaa tcctaacctt
     2881 tcgcatacct tactatgtgg gacccctggc ccgagggaac tctcggttcg catggatgac
     2941 aagaaagtcc gaagaaacga ttactccatg gaattttgag gaagttgtcg ataaaggtgc
     3001 gtcagctcaa tcgttcatcg agaggatgac caactttgac aagaatttac cgaacgaaaa
     3061 agtattgcct aagcacagtt tactttacga gtatttcaca gtgtacaatg aactcacgaa
     3121 agttaagtat gtcactgagg gcatgcgtaa acccgccttt ctaagcggag aacagaagaa
     3181 agcaatagta gatctgttat tcaagaccaa ccgcaaagtg acagttaagc aattgaaaga
     3241 ggactacttt aagaaaattg aatgcttcga ttctgtcgag atctccgggg tagaagatcg
     3301 atttaatgcg tcacttggta cgtatcatga cctcctaaag ataattaaag ataaggactt
     3361 cctggataac gaagagaatg aagatatctt agaagatata gtgttgactc ttaccctctt
     3421 tgaagatcgg gaaatgattg aggaaagact aaaaacatac gctcacctgt tcgacgataa
     3481 ggttatgaaa cagttaaaga ggcgtcgcta tacgggctgg ggacgattgt cgcggaaact
     3541 tatcaacggg ataagagaca agcaaagtgg taaaactatt ctcgattttc taaagagcga
     3601 cggcttcgcc aataggaact ttatgcagct gatccatgat gactctttaa ccttcaaaga
     3661 ggatatacaa aaggcacagg tttccggaca aggggactca ttgcacgaac atattgcgaa
     3721 tcttgctggt tcgccagcca tcaaaaaggg catactccag acagtcaaag tagtggatga
     3781 gctagttaag gtcatgggac gtcacaaacc ggaaaacatt gtaatcgaga tggcacgcga
     3841 aaatcaaacg actcagaagg ggcaaaaaaa cagtcgagag cggatgaaga gaatagaaga
     3901 gggtattaaa gaactgggca gccagatctt aaaggagcat cctgtggaaa atacccaatt
     3961 gcagaacgag aaactttacc tctattacct acaaaatgga agggacatgt atgttgatca
     4021 ggaactggac ataaaccgtt tatctgatta cgacgtcgat cacattgtac cccaatcctt
     4081 tttgaaggac gattcaatcg acaataaagt gcttacacgc tcggataaga accgagggaa
     4141 aagtgacaat gttccaagcg aggaagtcgt aaagaaaatg aagaactatt ggcggcagct
     4201 cctaaatgcg aaactgataa cgcaaagaaa gttcgataac ttaactaaag ctgagagggg
     4261 tggcttgtct gaacttgaca aggccggatt tattaaacgt cagctcgtgg aaacccgcca
     4321 aatcacaaag catgttgcac agatactaga ttcccgaatg aatacgaaat acgacgagaa
     4381 cgataagctg attcgggaag tcaaagtaat cactttaaag tcaaaattgg tgtcggactt
     4441 cagaaaggat tttcaattct ataaagttag ggagataaat aactaccacc atgcgcacga
     4501 cgcttatctt aatgccgtcg tagggaccgc actcattaag aaatacccga agctagaaag
     4561 tgagtttgtg tatggtgatt acaaagttta tgacgtccgt aagatgatcg cgaaaagcga
     4621 acaggagata ggcaaggcta cagccaaata cttcttttat tctaacatta tgaatttctt
     4681 taagacggaa atcactctgg caaacggaga gatacgcaaa cgacctttaa ttgaaaccaa
     4741 tggggagaca ggtgaaatcg tatgggataa gggccgggac ttcgcgacgg tgagaaaagt
     4801 tttgtccatg ccccaagtca acatagtaaa gaaaactgag gtgcagaccg gagggttttc
     4861 aaaggaatcg attcttccaa aaaggaatag tgataagctc atcgctcgta aaaaggactg
     4921 ggacccgaaa aagtacggtg gcttcgatag ccctacagtt gcctattctg tcctagtagt
     4981 ggcaaaagtt gagaagggaa aatccaagaa actgaagtca gtcaaagaat tattggggat
     5041 aacgattatg gagcgctcgt cttttgaaaa gaaccccatc gacttccttg aggcgaaagg
     5101 ttacaaggaa gtaaaaaagg atctcataat taaactacca aagtatagtc tgtttgagtt
     5161 agaaaatggc cgaaaacgga tgttggctag cgccggagag cttcaaaagg ggaacgaact
     5221 cgcactaccg tctaaatacg tgaatttcct gtatttagcg tcccattacg agaagttgaa
     5281 aggttcacct gaagataacg aacagaagca actttttgtt gagcagcaca aacattatct
     5341 cgacgaaatc atagagcaaa tttcggaatt cagtaagaga gtcatcctag ctgatgccaa
     5401 tctggacaaa gtattaagcg catacaacaa gcacagggat aaacccatac gtgagcaggc
     5461 ggaaaatatt atccatttgt ttactcttac caacctcggc gctccagccg cattcaagta
     5521 ttttgacaca acgatagatc gcaaacgata cacttctacc aaggaggtgc tagacgcgac
     5581 actgattcac caatccatca cgggattata tgaaactcgg atagatttgt cacagcttgg
     5641 gggtgactct ggtggttcta ctaatctgtc agatattatt gaaaaggaga ccggtaagca
     5701 actggttatc caggaatcca tcctcatgct cccagaggag gtggaagaag tcattgggaa
     5761 caagccggaa agcgatatac tcgtgcacac cgcctacgac gagagcaccg acgagaatgt
     5821 catgcttctg actagcgacg cccctgaata caagccttgg gctctggtca tacaggatag
     5881 caacggtgag aacaagatta agatgctctc tggtggttct cccaagaaga agaggaaagt
     5941 ctaaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc ctcgactgtg
     6001 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6061 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     6121 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     6181 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     6241 agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6301 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6361 aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6421 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6481 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6541 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6601 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6661 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6721 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6781 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6841 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6901 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6961 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7021 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7081 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7141 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7201 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7261 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7321 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7381 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7441 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7501 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7561 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7621 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7681 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7741 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7801 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7861 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7921 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7981 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8041 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8101 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8161 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8221 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8281 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8341 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8401 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     8461 gatcgatctc ccgatcccct agggtcgact ctcagtacaa tctgctctga tgccgcatag
     8521 ttaagccagt atctgctccc tgcttgtgtg ttg
//