LOCUS       sequence-452470-        4166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CyOFP1, for MuSIC barcode construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4166)
  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 4166)
  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-CyOFP1
FEATURES             Location/Qualifiers
     source          1..4166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..176
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      220..675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(307..326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     517..538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    682..803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(693..712)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     747..766
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(882..1583)
                     /codon_start=1
                     /product="orange-red fluorescent protein that is excitable 
                     by cyan light and suitable as an acceptor for blue-emitting
                     luciferase (Chu et al., 2016)"
                     /label=CyOFP1
                     /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTN
                     RIKVVEGGPLPFAFDILATHFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG
                     VLTATQDTSLQDGELIYNVKVRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD
                     KALKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
     regulatory      1580..1589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1637..1840)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1666..1686)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1841..2144
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2325..2913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2405..2424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2972..3019
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3044..3063
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3251..4045)
                     /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(3251..4045)
                     /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(3362..3381)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3972..3991
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4080..4166,1..110))
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(join(4088..4166,1..132))
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(4094..4166,1..63)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
       61 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
      121 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      181 ctggttcttt ccgcctcagg actcttcctt taagtactga cgcgccctgt agcggcgcat
      241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      661 taacgcttac aatttacgcc ttaagataca ttgatgagtt tggacaaacc acaactagaa
      721 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
      781 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcagaatt
      841 cattatctag aattcgtctt caattatgaa gacatggagc ccttgtacag ctcgtccatg
      901 cctccaccca ggttggagta tctggccaca gcgtgctcgt actgctcgac gtaagtctcg
      961 ttgtcggcct ccttgattct ttccagtctg cggtccacgt agtggacgcc gggcatcttc
     1021 acgggtttct tggacttgta tgtggtcttg aagttgacgt gtaagtgtcc acctcccacg
     1081 agcttcaggg ccttgtcgca tcttccttcc agacctccgt cagctgggta catggtttcg
     1141 gtgcttggtt cccagcccag tgttttcttc tgcatcacgg ggccgttggc tgggaagttc
     1201 acccctctga ccttgacgtt gtagatgagc tcgccgtcct ggaggctggt gtcctgggtg
     1261 gcggtcagca cgcccccgtc ctcgaacacc atgactctct cccatgtgaa gccctcaggg
     1321 aaggactgct taaaataatc ggggaggtcg gcggggtact tgatgaacac cttgctcccg
     1381 tacataaagt gggtggccag gatgtcgaat gcgaacggca gggggcctcc ctccaccacc
     1441 ttgatcctgt tggtctgctt gccctcgtag ggcttgccct ccccttcgtg ggtgcacttg
     1501 aactggtggc cgttcacgct gccttccagg tacagcttgc ttctcatgtt ctccttgatc
     1561 agctcctcgc ccttgctcac catggtggcg gcggatccat ttcgataagc cagtaagcac
     1621 gaggctgatc agcggtagct ctgcttatat agacctccca ccgtacacgc ctaccgccca
     1681 tttgcgtcaa tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc
     1741 aaaacaaact cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc
     1801 gctatccacg cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta
     1861 atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc
     1921 aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt
     1981 gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag
     2041 tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt
     2101 tgggcggtca gccaggcggg ccatttaccg taagttatgt aacggctagc cggaactcca
     2161 tatatgggct atgaactaat gaccccgtaa ttgattacta ttaataacta atgcatggcg
     2221 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2281 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2341 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2401 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2461 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2521 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2581 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2641 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2701 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2761 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2821 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2881 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2941 tctgacgctc agtggaacga accgcgttta tgaacaaacg acccaacacc gtgcgtttta
     3001 ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt
     3061 tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     3121 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     3181 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     3241 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     3301 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     3361 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     3421 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     3481 catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt
     3541 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     3601 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     3661 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3721 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3781 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     3841 acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga
     3901 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     3961 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     4021 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct
     4081 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
     4141 gaggcggcct cggcctctgc ataaat
//
