LOCUS       sequence-454230-        4169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mBeRFP, for MuSIC barcode construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4169)
  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 4169)
  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-mBeRFP
FEATURES             Location/Qualifiers
     source          1..4169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    359..480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(396..415)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     450..469
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(487..942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(624..645)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     836..855
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(986..1006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1030..1240
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        1052..1248
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1099..1234
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1283..2077
                     /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             1283..2077
                     /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(1337..1356)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1947..1966
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2265..2284)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2309..2356
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2415..3003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2904..2923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3184..3487
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3488..3691
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3642..3662
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3739..3748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cggttgggaa gcctccacag agatgctgta cccagctgac ggaggtctgg aaggaagaga
       61 ctacatggct ctgaagctcg tgggcggggg ccacctgatc tgcaacgcca agaccacata
      121 cagatccaag aaacccgcta agaacctcaa gatgcccggc gtctactatg tggacaggag
      181 actggaaaga atcaaggagg ccgacaaaga gacaagcgtc gagcagcacg aggtggctgt
      241 ggccagatac tgcgacctcc ctagcaaact ggggcacaaa ggctccatgt cttcataatt
      301 gaagacgaat tctagataat gaattctgaa acataaaatg aatgcaattg ttgttgttaa
      361 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
      421 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
      481 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
      541 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      601 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      661 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      721 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      781 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      841 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      901 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcagtactt aaaggaagag
      961 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     1021 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     1081 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     1141 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     1201 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     1261 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     1321 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     1381 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     1441 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     1501 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     1561 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     1621 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     1681 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     1741 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     1801 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     1861 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     1921 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     1981 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     2041 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     2101 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     2161 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     2221 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     2281 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     2341 gttgggtcgt ttgttcataa acgcggttcg ttccactgag cgtcagaccc cgtagaaaag
     2401 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2461 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2521 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2581 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2641 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2701 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2761 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2821 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2881 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2941 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3001 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3061 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag
     3121 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccggct
     3181 agccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     3241 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     3301 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     3361 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     3421 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     3481 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     3541 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     3601 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     3661 tgtacggtgg gaggtctata taagcagagc taccgctgat cagcctcgtg cttactggct
     3721 tatcgaaatg gatccgccgc caccatggtg tctaagggcg aagagctgat taaggagaac
     3781 atgcatatga agctgtacat ggagggcacc gtgaacaacc accacttcaa gtgcacatcc
     3841 gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc
     3901 ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg cagcaaaacc
     3961 ttcatcaacc acacccaggg catccccgac ttctttaagc agtccttccc tgagggcttc
     4021 acatgggaga gatccaccac atacgaggac gggggcgtgc tgaccgctac ccaggacacc
     4081 agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa cttcccatcc
     4141 aacggccctg tgatgcagaa gaaaacact
//
