LOCUS       sequence-454228-        4175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mTagBFP2, for MuSIC barcode construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4175)
  AUTHORS   Lu X, Pritko DJ, Abravanel ME, Huggins JR, Ogunleye O, Biswas T, 
            Ashy KC, Woods SK, Livingston MWT, Blenner MA, Birtwistle MR
  TITLE     Genetically Encoded Fluorescence Barcodes Allow for Single-Cell 
            Analysis via Spectral Flow Cytometry.
  JOURNAL   ACS Synth Biol. 2025 May 16;14(5):1533-1548. doi: 
            10.1021/acssynbio.4c00807. Epub 2025 May 6.
  PUBMED    40326708
REFERENCE   2  (bases 1 to 4175)
  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"; doi: 
            "10.1021/acssynbio.4c00807"; journalName: "ACS Synth Biol"; date: 
            "2025-05-16- 16"; volume: "14"; issue: "5"; pages: "1533-1548"
COMMENT     Sequence Label: pR-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..4175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(64..858)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(64..858)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(175..194)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     785..804
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(893..1089)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(901..1111)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      907..1042
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1135..1155
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1199..1654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1286..1305)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1496..1517
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1661..1782
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1672..1691)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1726..1745
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1861..2571)
                     /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"
     regulatory      2568..2577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2625..2828)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2654..2674)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2829..3132
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3313..3901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3393..3412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3960..4007
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4032..4051
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt catttcgaac cccagagtcc
       61 cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc
      121 gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc
      181 acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat
      241 gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt
      301 cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg
      361 cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg
      421 agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc
      481 aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag
      541 gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc
      601 ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag
      661 ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag
      721 aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg
      781 ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa
      841 tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcga tctttgcaaa
      901 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
      961 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
     1021 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
     1081 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
     1141 actgacacac attccacagc tggttctttc cgcctcagga ctcttccttt aagtactgac
     1201 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1261 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1321 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1381 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1441 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1501 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1561 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1621 gcgaatttta acaaaatatt aacgcttaca atttacgcct taagatacat tgatgagttt
     1681 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1741 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1801 cattttatgt ttcagaattc attatctaga attcgtcttc aattatgaag acatggagcc
     1861 attaagcttg tgccccagtt tgctagggag gtcgcagtat ctggccactg ccacctcgtg
     1921 ctgctcgacg tatgtctcgt tgttggcctc cttgattctt tccagtctgt agtccacata
     1981 gtagacgcca ggcatcttga ggttcttagc gggtttcttg gatctatatg tggtcttggc
     2041 gtttgcgatc agatggctcc cgcccacgag cttcagggcc atgtcgtttc tgccttccag
     2101 gccgccgtca gcggggtaca gtgtctcggt gaaggcctcc cagccgagtg ttttcttctg
     2161 catcacaggg ccgttggatg tgaagttcac ccctctgatc ttgacgttgt agatgaggca
     2221 gccgtcctgg aggctggtgt cctgggtagc ggtcagcacg cccccatctt cgtatgtggt
     2281 gactctctcc catgtgaagc cctcagggaa ggactgcttg aagaagtcgg ggatgccctg
     2341 ggtgtggttg atgaaggtct tgctgccgta gaggaagcta gtagccagga tgtcgaaggc
     2401 gaaggggaga gggccgccct cgaccacctt gattctcatg gtctgggtgc cctcgtaggg
     2461 cttgccttcg ccctcggatg tgcacttgaa gtgatggttg tccacggtgc cctccatgta
     2521 cagcttcatg tgcatgttct ccttaatcag ctcttcgccc ttagacacca tggtggcggc
     2581 ggatccattt cgataagcca gtaagcacga ggctgatcag cggtagctct gcttatatag
     2641 acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat
     2701 tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg
     2761 agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac
     2821 cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc
     2881 cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata
     2941 gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa
     3001 tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc
     3061 attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta
     3121 agttatgtaa cggctagccg gaactccata tatgggctat gaactaatga ccccgtaatt
     3181 gattactatt aataactaat gcatggcggt aatacggtta tccacagaat caggggataa
     3241 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3301 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3361 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3421 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3481 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3541 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3601 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3661 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3721 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3781 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3841 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3901 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaac cgcgtttatg
     3961 aacaaacgac ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt
     4021 tccttccggt attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa
     4081 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     4141 gaagcccaac ctttcataga aggcggcggt ggaat
//
