LOCUS       sequence-252310-        7157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mTagBFP2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7157)
  AUTHORS   Ochiai H, Hayashi T, Umeda M, Yoshimura M, Harada A, Shimizu Y, 
            Nakano K, Saitoh N, Liu Z, Yamamoto T, Okamura T, Ohkawa Y, Kimura 
            H, Nikaido I
  TITLE     Genome-wide kinetic properties of transcriptional bursting in mouse 
            embryonic stem cells.
  JOURNAL   Sci Adv. 2020 Jun 17;6(25):eaaz6699. doi: 10.1126/sciadv.aaz6699. 
            eCollection 2020 Jun.
  PUBMED    32596448
REFERENCE   2  (bases 1 to 7157)
  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: "Sci Adv."; 
            date: "2020-06-17"; volume: "6(25):eaaz6699. doi"; pages: " 
            10.1126/sciadv.aaz6699. eCollection 2020 Jun"
COMMENT     Sequence Label: pCAG-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..7157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          161..1177
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1101..1123
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1185..1204
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     1275..1294
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1275..1293
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          2202..2431
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2414..2433
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2504..3078,3079..3081,3082..3090)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2671..2688)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2898..2917
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3113..3916
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3176..3195)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3786..3805
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4046..4167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4083..4102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4269..4285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4269..4285)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4282..4304)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4293..4309
                     /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(4317..4323,4324..4341,4342..4347))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4362..4383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4500..4517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4671..5259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4751..4770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5430..6221,6222..6290))
                     /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     6053..6072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6291..6395)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6470..6489)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6661..7040
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(7042..7157,1..160)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggggggcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
       61 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      121 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
      181 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
      241 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
      301 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
      361 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
      421 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
      481 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
      541 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
      601 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
      661 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc
      721 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga
      781 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
      841 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
      901 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
      961 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     1021 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
     1081 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1141 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     1201 tgtgctgtct catcattttg gcaaagagct ctctggctaa ctagagaacc cactgcttac
     1261 tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta tcgacctcgg
     1321 atcgtcgcta gcgatatcgg cgcgccagca tttaaatctg tacagaccgg tgaattcacc
     1381 atggtgtcca agggtgagga actcataaag gaaaacatgc acatgaaatt atacatggag
     1441 ggcacagtag acaatcatca ttttaagtgc acctccgaag gcgaaggcaa gccctacgag
     1501 ggcactcaga caatgagaat caaggtggtc gaggggggtc cgctaccctt tgccttcgac
     1561 attttggcta ccagttttct gtacgggagc aagactttca taaaccacac ccaggggatt
     1621 cctgattttt ttaaacagag ttttcctgag ggctttacat gggaacgggt gacgacttac
     1681 gaggacggag gagtcctgac cgcgacccag gatacctcgt tgcaggacgg atgtttaatt
     1741 tacaatgtaa agatccgggg ggtaaatttc acgtccaatg gtcctgttat gcagaaaaaa
     1801 actctggggt gggaagcctt tacagagact ctgtaccccg ctgacggcgg actggagggt
     1861 aggaacgata tggctttgaa actcgtgggg ggttcacact taatagcaaa cgctaagact
     1921 acctatagaa gcaaaaagcc tgctaagaac ttgaagatgc ctggtgtata ctacgtggat
     1981 tacaggctgg aacgtatcaa agaggcaaac aacgaaacat acgtggaaca acatgaggtt
     2041 gccgtcgcta ggtactgcga tttgccatct aaactcggtc ataagttgaa ccctgcaggc
     2101 ggatccgcgt ttaaactcga ggttaattaa tgagcggccg catagataac tgatccagtg
     2161 tgctggaatt aattcgctgt ctgcgagggc cagctgttgg ggtgagtact ccctctcaaa
     2221 agcgggcatg acttctgcgc taagattgtc agtttccaaa aacgaggagg atttgatatt
     2281 cacctggccc gcggtgatgc ctttgagggt ggccgcgtcc atctggtcag aaaagacaat
     2341 ctttttgttg tcaagcttga ggtgtggcag gcttgagatc tggccataca cttgagtgac
     2401 aatgacatcc actttgcctt tctctccaca ggtgtccact cccaggtcca actgcaggtc
     2461 gagcatgcat ctagggcggc caattccgcc cctctccccc ccccccctct ccctcccccc
     2521 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     2581 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     2641 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     2701 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     2761 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     2821 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     2881 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     2941 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     3001 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     3061 gaaaaacacg atgataagct tgccacaacc cgggataatt cctgcagcca atatgggatc
     3121 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3181 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3241 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3301 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3361 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3421 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3481 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3541 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3601 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3661 cggcgatgat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3721 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3781 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3841 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3901 tgacgagttc ttctgagggg atcaattctc tagataactg atcataatca gccataccac
     3961 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4021 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4081 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4141 tttgtccaaa ctcatcaatg tatcttaacg cgtcgagtgc attctagttg tggtttgtcc
     4201 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4261 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4321 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4381 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4441 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4501 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4561 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4621 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4681 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4741 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4801 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4861 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4921 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4981 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5041 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5101 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5161 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5221 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5281 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5341 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5401 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5461 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5521 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5581 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5641 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5701 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5761 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5821 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5881 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5941 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6001 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6061 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6121 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6181 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6241 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6301 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6361 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6421 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6481 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6541 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     6601 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     6661 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6721 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6781 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6841 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6901 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6961 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     7021 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     7081 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     7141 tgggggcggg ggggggg
//
