LOCUS       sequence-452450-        4172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of LSSmOrange, 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-LSSmOrange
FEATURES             Location/Qualifiers
     source          1..4172
                     /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..1589)
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     regulatory      1586..1595
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1643..1846)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1672..1692)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1847..2150
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2331..2919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2411..2430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2978..3025
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3050..3069
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3257..4051)
                     /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(3257..4051)
                     /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(3368..3387)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3978..3997
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4086..4172,1..110))
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(join(4094..4172,1..132))
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(4100..4172,1..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 ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg
      961 ttgtgtgagg tgatgtccaa cttgatgtcg acgatgtagg cgcctggcaa ctgcacgggc
     1021 ttcttggcct tgtaggtggt cttgacctcg gaggtgtagt ggccgccgtc cttcagcttc
     1081 agcctgagct tgtcctcgcc cttcagtgcg ccgtcctcgg ggtacatccg ttcagatgag
     1141 gcttccatgc ccatggtttt cttctgcatt acgggtccgt cggaggggaa gttggtgccg
     1201 cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg agtcacggtc
     1261 accacgccgc catcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac
     1321 agcttgaggt agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtaggtg
     1381 aactgaggtg acaatatgtc ccaggcgaaa ggcagtggtc cacccttggt aaccttcagc
     1441 ttaacggtct gaaagccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg
     1501 tggccgttca cggagccctc catgcgcacc ttgaagcgca tgaactcctt gatgatggcc
     1561 atgttattct cctcgccctt gctcaccatg gtggcggcgg atccatttcg ataagccagt
     1621 aagcacgagg ctgatcagcg gtagctctgc ttatatagac ctcccaccgt acacgcctac
     1681 cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc cgttgatttt
     1741 ggtgccaaaa caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt
     1801 caaaccgcta tccacgccca ttgatgtact gccaaaaccg catcaccatg gtaatagcga
     1861 tgactaatac gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat
     1921 aatgccaggc gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat
     1981 acacttgatg tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat
     2041 ggaaagtccc tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg
     2101 ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg gctagccgga
     2161 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactaatgc
     2221 atggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2281 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2341 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2401 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2461 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2521 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2581 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2641 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2701 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2761 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2821 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2881 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2941 acggggtctg acgctcagtg gaacgaaccg cgtttatgaa caaacgaccc aacaccgtgc
     3001 gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc
     3061 cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc
     3121 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     3181 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     3241 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     3301 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     3361 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     3421 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     3481 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     3541 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     3601 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     3661 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     3721 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     3781 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     3841 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     3901 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     3961 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     4021 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     4081 cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     4141 gaggccgagg cggcctcggc ctctgcataa at
//
