LOCUS       sequence-454234-        4172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mOrange2, for MuSIC barcode construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4172)
  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 4172)
  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-mOrange2
FEATURES             Location/Qualifiers
     source          1..4172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(109..903)
                     /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(109..903)
                     /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(220..239)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     830..849
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(938..1134)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(946..1156)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      952..1087
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1180..1200
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1244..1699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1331..1350)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1541..1562
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1706..1827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1717..1736)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1771..1790
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1906..2613)
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     regulatory      2610..2619
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2667..2870)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2696..2716)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2871..3174
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3355..3943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3435..3454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4002..4049
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4074..4093
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg
       61 gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc
      121 gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac
      181 gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc
      241 tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg
      301 gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc
      361 gccgtcgggc atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg
      421 ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc
      481 gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg
      541 ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag
      601 atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc
      661 gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc
      721 ttgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg
      781 cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata
      841 gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat
      901 catgcgaaac gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa
      961 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     1021 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     1081 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     1141 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
     1201 acagctggtt ctttccgcct caggactctt cctttaagta ctgacgcgcc ctgtagcggc
     1261 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1321 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1381 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1441 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1501 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1561 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1621 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1681 atattaacgc ttacaattta cgccttaaga tacattgatg agtttggaca aaccacaact
     1741 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1801 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1861 aattcattat ctagaattcg tcttcaatta tgaagacatg gagcccttgt acagctcgtc
     1921 catgccgccg gtggagtggc ggccctcggc gcgttcgtac tgttccacga tggtgtagtc
     1981 ctcgttgtgg gaggtgatgt ccaacttgat gtcgacgatg taggcgccgg gcagctgcac
     2041 gggcttcttg gccttgtagg tggtcttgac ctcggaggtg tagtggccgc cgtccttcag
     2101 cttcagcctc atcttgatct tgcccttcag ggcaccgtcc tcggggtaca tccgctcgga
     2161 ggaggcctcc cagcccatgg ttttcttctg catcacgggg ccgtcggagg ggaagttggt
     2221 gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc agggaggagt cctgggtcac
     2281 ggtcaccacg ccgccgtcct cgtagttcat cacgcgctcc cacttgaagc cctcggggaa
     2341 ggacagcttg aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct tggagccgta
     2401 ggtgaaatga ggggacagga tgtcccaggc gaagggcagg gggccaccct tggtcacctt
     2461 cagcttagcg gtctgaaagc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa
     2521 ctcgtggccg ttcacggagc cctccatgcg caccttgaag cgcatgaact ccttgatgat
     2581 ggccatgtta ttctcctcgc ccttgctcac catggtggcg gcggatccat ttcgataagc
     2641 cagtaagcac gaggctgatc agcggtagct ctgcttatat agacctccca ccgtacacgc
     2701 ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga
     2761 ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt
     2821 gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata
     2881 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     2941 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     3001 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     3061 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     3121 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacggctagc
     3181 cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta
     3241 atgcatggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3301 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3361 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3421 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3481 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3541 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3601 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3661 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3721 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3781 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3841 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3901 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3961 ttctacgggg tctgacgctc agtggaacga accgcgttta tgaacaaacg acccaacacc
     4021 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     4081 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     4141 gcgctggagg atcatccagc cggcgtcccg ga
//
