LOCUS       sequence-435546-        4742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mTagBFP2 expression in mammalian cells. Reporter plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4742)
  AUTHORS   Barykina NV, Carey EM, Oliinyk OS, Nimmerjahn A, Verkhusha VV
  TITLE     Destabilized near-infrared fluorescent nanobodies enable 
            background-free targeting of GFP-based biosensors for imaging and 
            manipulation.
  JOURNAL   Nat Commun. 2024 Sep 6;15(1):7788. doi: 10.1038/s41467-024-51857-x.
  PUBMED    39242569
REFERENCE   2  (bases 1 to 4742)
  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: "Nat 
            Commun."; date: "2024-09-6"; volume: "15(1):7788. doi"; pages: " 
            10.1038/s41467-024-51857-x"
COMMENT     Sequence Label: pGP-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..4742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..33
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     15..32
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     intron          1626..1691
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1821..1841
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2266..2400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2303..2322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2357..2376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2410..2429)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2411..2429)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2516..2533)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2687..3275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2767..2786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3446..4237,4238..4306))
                     /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     4069..4088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4307..4411)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4479..4497
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4535..4557)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4657..4676
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atcatacaca tacgatttag gtgacactat agaactcgag cagctgaagc ttcggagtac
       61 tgtcctccga gcggagtact gtcctccgag cggagtactg tcctccgagt cgactctagc
      121 ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagtcga
      181 ctctagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggagt actgtcctcc
      241 ggctagtagc ggagactcta gagggtatat aatggatccc cgggtacccg gggatccacc
      301 atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtggac
      361 aaccatcact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
      421 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctact
      481 agcttcctct acggcagcaa gaccttcatc aaccacaccc agggcatccc cgacttcttc
      541 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
      601 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
      661 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
      721 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
      781 gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg ccaagaccac atatagatcc
      841 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
      901 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
      961 tactgcgacc tccctagcaa actggggcac aagtaaagcg gccgcgactc tagaattccg
     1021 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     1081 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     1141 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     1201 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     1261 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     1321 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     1381 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     1441 gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggttg
     1501 ctacgcctga ataagtgata ataagcggat gaatggcaga aattcgccgg atctttgtga
     1561 aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct
     1621 ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
     1681 gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta
     1741 atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg
     1801 actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc
     1861 cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct
     1921 ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat
     1981 attctgtaac ctttataagt aggcataaca gttataatca taacatactg ttttttctta
     2041 ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta
     2101 gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag
     2161 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2221 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2281 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2341 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     2401 tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat
     2461 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2521 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2581 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2641 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2701 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2761 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2821 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2881 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2941 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3001 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3061 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3121 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3181 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3241 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3301 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3361 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3421 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3481 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3541 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3601 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3661 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3721 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3781 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3841 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3901 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3961 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4021 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4081 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4141 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4201 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4261 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4321 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4381 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4441 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     4501 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     4561 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     4621 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     4681 gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat
     4741 gt
//
