LOCUS       sequence-233517-        7150 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 7150)
  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 7150)
  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..7150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..892
                     /label=chicken beta-actin promoter
     intron          893..1901
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1825..1847
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1999..2018
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1999..2017
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          2926..3155
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     3138..3157
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3228..3801
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3395..3412)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3622..3641
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3837..4640
                     /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(3900..3919)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4510..4529
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4770..4891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4807..4826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4993..5009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4993..5009)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5006..5028)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5017..5033
                     /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(5041..5047,5048..5065,5066..5071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5086..5107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5224..5241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5395..5983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5475..5494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6154..6945,6946..7014))
                     /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     6777..6796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7015..7119)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catgggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
      661 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      721 gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc
      781 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      841 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      901 tgcgttgcct tcgccccgtg ccccgctccg cgccgcctcg cgccgcccgc cccggctctg
      961 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
     1021 ttagcgcttg gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa gccttaaagg
     1081 gctccgggag ggccctttgt gcggggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
     1141 gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc tgtgagcgct gcgggcgcgg
     1201 cgcggggctt tgtgcgctcc gcgtgtgcgc gaggggagcg cggccggggg cggtgccccg
     1261 cggtgcgggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     1321 gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc ccctccccga
     1381 gttgctgagc acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg cgcggggctc
     1441 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     1501 gccggggagg gctcggggga ggggcgcggc ggccccggag cgccggcggc tgtcgaggcg
     1561 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     1621 tgtcccaaat ctggcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1681 cgggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     1741 ccgcgccgcc gtccccttct ccatctccag cctcggggct gccgcagggg gacggctgcc
     1801 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1861 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1921 ttgttgtgct gtctcatcat tttggcaaag agctctctgg ctaactagag aacccactgc
     1981 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt ggtatcgacc
     2041 tcggatcgtc gctagcgata tcggcgcgcc agcatttaaa tctgtacaga ccggtgaatt
     2101 caccatggtg tccaagggtg aggaactcat aaaggaaaac atgcacatga aattatacat
     2161 ggagggcaca gtagacaatc atcattttaa gtgcacctcc gaaggcgaag gcaagcccta
     2221 cgagggcact cagacaatga gaatcaaggt ggtcgagggg ggtccgctac cctttgcctt
     2281 cgacattttg gctaccagtt ttctgtacgg gagcaagact ttcataaacc acacccaggg
     2341 gattcctgat ttttttaaac agagttttcc tgagggcttt acatgggaac gggtgacgac
     2401 ttacgaggac ggaggagtcc tgaccgcgac ccaggatacc tcgttgcagg acggatgttt
     2461 aatttacaat gtaaagatcc ggggggtaaa tttcacgtcc aatggtcctg ttatgcagaa
     2521 aaaaactctg gggtgggaag cctttacaga gactctgtac cccgctgacg gcggactgga
     2581 gggtaggaac gatatggctt tgaaactcgt ggggggttca cacttaatag caaacgctaa
     2641 gactacctat agaagcaaaa agcctgctaa gaacttgaag atgcctggtg tatactacgt
     2701 ggattacagg ctggaacgta tcaaagaggc aaacaacgaa acatacgtgg aacaacatga
     2761 ggttgccgtc gctaggtact gcgatttgcc atctaaactc ggtcataagt tgaaccctgc
     2821 aggcggatcc gcgtttaaac tcgaggttaa ttaatgagcg gccgcataga taactgatcc
     2881 agtgtgctgg aattaattcg ctgtctgcga gggccagctg ttggggtgag tactccctct
     2941 caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga
     3001 tattcacctg gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga
     3061 caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag
     3121 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca
     3181 ggtcgagcat gcatctaggg cggccaattc cgcccctctc cccccccccc ttttccctcc
     3241 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3301 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3361 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3421 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3481 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3541 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3601 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3661 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3721 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3781 ctttgaaaaa cacgatgata agcttgccac aacccgggat aattcctgca gccaatatgg
     3841 gatcggccat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3901 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3961 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4021 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4081 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4141 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4201 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4261 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4321 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4381 ccgacggcga tgatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4441 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4501 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4561 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4621 ttcttgacga gttcttctga ggggatcaat tctctagata actgatcata atcagccata
     4681 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4741 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4801 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4861 gtggtttgtc caaactcatc aatgtatctt aacgcgtcga gtgcattcta gttgtggttt
     4921 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4981 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5041 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5101 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5161 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5221 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5281 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5401 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5461 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5521 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5581 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5641 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5701 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5761 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5821 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5881 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6001 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6061 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6121 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6601 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6961 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7021 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7081 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7141 acctgacgtc
//
