LOCUS       sequence-384555-        5594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  It expresses HA-fused mouse Set2 isoform protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5594)
  AUTHORS   Molla MR, Shimizu A, Komeno M, Rahman NIA, Soh JEC, Nguyen LKC, Khan
            MR, Tesega WW, Chen S, Pang X, Tanaka-Okamoto M, Takashima N, Sato 
            A, Suzuki T, Ogita H
  TITLE     Vascular smooth muscle RhoA counteracts abdominal aortic aneurysm 
            formation by modulating MAP4K4 activity.
  JOURNAL   Commun Biol. 2022 Oct 7;5(1):1071. doi: 10.1038/s42003-022-04042-z.
  PUBMED    36207400
REFERENCE   2  (bases 1 to 5594)
  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: "Commun 
            Biol."; date: "2022-10-7"; volume: "5(1):1071. doi"; pages: " 
            10.1038/s42003-022-04042-z"
COMMENT     Sequence Label: pCMV-HA Set2
FEATURES             Location/Qualifiers
     source          1..5594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     29..46
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    163..184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(199..204,205..222,223..229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    237..253
                     /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     242..264
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     261..277
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     261..277
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        306..685
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        686..889
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     840..860
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     886..910
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1120..1138
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1120..1137
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1151..1170
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1225..1314
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     regulatory      2186..2195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2199..2333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2236..2255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2290..2309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2402..2759
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2402..2593
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2425..2445)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2610..2745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2779..2796)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2779..2795)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2788..2810)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3008..3463
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3095..3114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3305..3326
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3480..3499)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3599..3621
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3659..3677)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3745..3849
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3850..3918,3919..4710)
                     /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(4068..4087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4881..5469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5370..5389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
       61 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      121 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      181 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      241 gagcggataa caatttcaca caggaaacag ctatgacatg attacgaatt aattcgagct
      301 cgcccgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      361 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      421 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      481 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      541 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      601 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      661 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      721 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      781 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      841 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
      901 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
      961 ccgatccagc ctccgcggcc gggaacggtg cattggaacg cggattcccc gtgccaagag
     1021 tgacgtaagt accgcctata gagtctatag gcccaccccc ttggcttcgt tagaacgcgg
     1081 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actatagaat
     1141 aacatccact ttgcctttct ctccacaggt gtccactccc aggtccaact gcacctcggt
     1201 tctatcgata tgggccgcat cttttaccca tacgatgttc ctgactatgc gggctatccc
     1261 tatgacgtcc cggactatgc aggatcctat ccatatgacg ttccagatta cgctgctcag
     1321 cgcgaattct ctgcgccgac ggccaaagcc agtaaaaagg agctcaactc caatcacgac
     1381 ggggccgacg agacctcaga aaaagaacag caagaagcaa ttgaacatat tgatgaagta
     1441 caaaatgaaa tagacagact taatgaacaa gccagtgagg aaattttgaa agtagaacaa
     1501 aaatataaca aactccgcca accatttttt cagaagaggt cagaattgat cgccaaaatc
     1561 ccaaattttt gggtaacaac atttgtcaac catccacaag tgtctgcact gcttggggag
     1621 gaggacgagg aggctctgca ttatttgacc agagttgaag tgacagaatt tgaagacatt
     1681 aaatcaggtt acagaataga tttttatttt gatgaaaatc cttactttga aaataaagtt
     1741 ctctccaaag aatttcatct gaacgagagt ggtgacccgt cttcaaagtc caccgaaatc
     1801 aaatggaaat ctggaaagga tttgacaaaa cgctcaagtc aaacgcaaaa taaggccagc
     1861 aggaagaggc agcacgaaga gccagagagc ttctttacct ggtttactga ccattctgac
     1921 gcaggtgctg atgagttagg agaggtcatc aaagatgaca tctggccaaa tcccttgcag
     1981 tactacctgg ttcccgacat ggatgatgaa gaaggagagg cagaagatga tgatgacgac
     2041 gacgaagagg aggaggggct ggaagatatt gatgaagaag gagatgagga tgaaggtgaa
     2101 gaagatgacg atgaggatga aggggaggaa ggagaggagg acgaaggcga ggatgattag
     2161 ctcgagtgac tgactgagaa gcttggccgc catggcccaa cttgtttatt gcagcttata
     2221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2281 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcgggaatt
     2341 aattcggcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
     2401 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2461 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2641 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     2701 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2761 agctgttaac agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2821 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2881 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
     2941 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3001 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3061 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3121 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3181 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3241 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3301 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     3361 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3421 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     3481 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3541 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3601 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3661 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3721 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3781 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3841 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3901 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3961 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4021 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4081 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4141 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4201 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4261 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4321 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4381 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4441 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4501 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4561 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4621 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4681 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4741 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4801 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4861 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4921 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4981 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5041 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5101 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5161 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5221 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5281 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5341 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5401 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5461 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5521 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5581 gagtgagctg atac
//
