LOCUS       sequence-380267-        5114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  It expresses HA-fused mouse Set1 isoform protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5114)
  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 5114)
  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 Set1
FEATURES             Location/Qualifiers
     source          1..5114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    110..131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(146..151,152..169,170..176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    184..200
                     /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     189..211
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     208..224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     208..224
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        253..632
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        633..836
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     787..807
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     833..857
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1067..1085
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1067..1084
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1098..1117
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1172..1261
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     regulatory      1653..1662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1666..1800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1703..1722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1757..1776
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1869..2226
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1869..2060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1892..1912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2077..2212
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2246..2263)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2246..2262)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2255..2277)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2475..2930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2562..2581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2772..2793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2947..2966)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3066..3088
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3126..3144)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3212..3316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3317..3385,3386..4177)
                     /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(3535..3554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4348..4936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4837..4856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5090..5107
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
       61 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      121 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      181 gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt acgaattaat
      241 tcgagctcgc ccgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      301 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      361 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      421 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      481 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      541 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      601 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      661 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      721 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      781 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
      841 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
      901 accgggaccg atccagcctc cgcggccggg aacggtgcat tggaacgcgg attccccgtg
      961 ccaagagtga cgtaagtacc gcctatagag tctataggcc cacccccttg gcttcgttag
     1021 aacgcggcta caattaatac ataaccttat gtatcataca catacgattt aggtgacact
     1081 atagaataac atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca
     1141 cctcggttct atcgatatgg gccgcatctt ttacccatac gatgttcctg actatgcggg
     1201 ctatccctat gacgtcccgg actatgcagg atcctatcca tatgacgttc cagattacgc
     1261 tgctcagcgc gaattcgccc cgaagcggca gtctgcgatc ctgcctcagc ccaagaaacc
     1321 cagaccggct gctgccccga agctggagga caagtcggcc tctcccggcc tgccgaaggg
     1381 agaaaaagaa cagcaagaag caattgaaca tattgatgaa gtacaaaatg aaatagacag
     1441 acttaatgaa caagccagtg aggaaatttt gaaagtagaa caaaaatata acaaactccg
     1501 ccaaccattt tttcagaaga ggtcagaatt gatcgccaaa atcccaaatt tttgggtaac
     1561 aacatttgtc aaccatccac aagtgtctgc actgcttggg gaggaggacg aaggcgagga
     1621 tgattagctc gagtgactga ctgagaagct tggccgccat ggcccaactt gtttattgca
     1681 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1741 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     1801 gggaattaat tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     1861 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     1921 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1981 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2041 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2101 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2161 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2221 tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     2281 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2341 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2401 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2461 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2521 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2581 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2641 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2701 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2761 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     2821 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2881 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2941 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     3001 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     3061 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     3121 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3181 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3241 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3301 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3361 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3421 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3481 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3541 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3601 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3661 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3721 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3781 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3841 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3901 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3961 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4021 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4081 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4141 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4201 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4261 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4321 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4381 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4441 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4501 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4561 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4621 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4681 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4741 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4801 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4861 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4921 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4981 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5041 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5101 gagcgaggaa gcgg
//
