LOCUS       sequence-306231-        7282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral (MSCV) expression of mTagBFP2 fluorescent protein .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7282)
  AUTHORS   Mann SE, Zhou Z, Landry LG, Anderson AM, Alkanani AK, Fischer J, 
            Peakman M, Mallone R, Campbell K, Michels AW, Nakayama M
  TITLE     Multiplex T Cell Stimulation Assay Utilizing a T Cell Activation 
            Reporter-Based Detection System.
  JOURNAL   Front Immunol. 2020 Apr 9;11:633. doi: 10.3389/fimmu.2020.00633. 
            eCollection 2020.
  PUBMED    32328071
REFERENCE   2  (bases 1 to 7282)
  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: "Front 
            Immunol."; date: "2020-04-9"; volume: "11:633. doi"; pages: " 
            10.3389/fimmu.2020.00633. eCollection 2020"
COMMENT     Sequence Label: pMIII-Neo-BFP
FEATURES             Location/Qualifiers
     source          1..7282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    64..405
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             472..888
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     818..840
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     856..872
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      903..912
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             909..1619
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     misc_feature    1654..2203
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1798..1815)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2025..2044
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2213..3016
                     /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
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2276..2295)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2886..2905
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3160..3674
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3836..3852
                     /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(3860..3866,3867..3884,3885..3890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3905..3926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4043..4060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4214..4802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4294..4313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4973..5764,5765..5833))
                     /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     5596..5615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5834..5938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6006..6024
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6062..6084)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6184..6203
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6397..6419
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6547..6566
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6766..7282
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tggaggttcc accgagattt ggagacccct gcccagggac caccgacccc cccgccggga
       61 ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt ttgtgccggc
      121 atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc
      181 cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac
      241 tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg
      301 tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
      361 tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc
      421 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
      481 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
      541 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
      601 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
      661 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
      721 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
      781 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
      841 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg aattcagatc
      901 tagccaccat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt
      961 acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc
     1021 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
     1081 ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc
     1141 agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca
     1201 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     1261 gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc
     1321 agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc
     1381 tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaacg
     1441 ccaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact
     1501 atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc
     1561 acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatt
     1621 gactcgagcg ggatcaattc cgcccccccc ctaacgttac tggccgaagc cgcttggaat
     1681 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     1741 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     1801 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     1861 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     1921 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     1981 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     2041 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     2101 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     2161 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatgaa ccatgggatc
     2221 ggccattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2281 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2341 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2401 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2461 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2521 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2581 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2641 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2701 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     2761 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     2821 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     2881 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     2941 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3001 tgacgagttc ttctgagtcg acctgcaggc atgcaagctt caggtagccg gctaacgtta
     3061 acaaccggta cctctagaac tatagctagc atgcgcaaat ttaaagcgct gatatcgata
     3121 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     3181 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     3241 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     3301 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     3361 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     3421 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     3481 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     3541 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     3601 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     3661 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
     3721 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
     3781 ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt
     3841 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3901 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3961 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4021 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4081 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4141 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4201 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4261 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4321 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4381 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4441 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4501 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4561 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4621 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4681 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4741 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4801 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4861 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4921 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4981 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5041 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5101 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5161 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5221 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5281 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5341 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5401 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5461 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5521 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5581 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5641 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5701 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5761 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5821 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5881 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5941 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     6001 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6061 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6121 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6181 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     6241 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     6301 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     6361 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     6421 cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     6481 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     6541 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     6601 ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt
     6661 ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc
     6721 atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac
     6781 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa
     6841 ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa
     6901 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     6961 agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     7021 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     7081 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg
     7141 ccagtcctcc gatagactgc gtcgcccggg tacccgtatt cccaataaag cctcttgctg
     7201 tttgcatccg aatcgtggac tcgctgatcc ttgggagggt ctcctcagat tgattgactg
     7261 cccacctcgg gggtctttca tt
//
