LOCUS       sequence-452466-        4160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mKate2, for MuSIC barcode construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4160)
  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 4160)
  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-mKate2
FEATURES             Location/Qualifiers
     source          1..4160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..38
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      115..124
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             121..816
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     polyA_signal    895..1016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(932..951)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     986..1005
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1023..1478)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1160..1181)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1372..1391
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(1522..1542)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1566..1776
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        1588..1784
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1635..1770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1819..2613
                     /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             1819..2613
                     /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(1873..1892)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2483..2502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2801..2820)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2845..2892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2951..3539
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3440..3459
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3720..4023
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(4024..4160,1..67)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
       61 cagagctacc gctgatcagc ctcgtgctta ctggcttatc gaaatggatc cgccgccacc
      121 atggtgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg
      181 aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
      241 accatgagaa tcaaggcggt cgagggcggc cctctcccct tcgccttcga catcctggct
      301 accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat ccccgacttc
      361 tttaagcagt ccttccccga gggcttcaca tgggagagag tcaccacata cgaagatggg
      421 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc
      481 aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa aacactcggc
      541 tgggaggcct ccaccgagac actgtacccc gctgacggcg gcctggaagg cagagccgac
      601 atggccctga agctcgtggg cgggggccac ctgatctgca acttaaagac cacatacaga
      661 tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga cagaaggctg
      721 gaaagaatca aggaggccga caaagagact tacgtcgagc agcacgaggt ggctgtggcc
      781 agatactgcg acctccctag caaactgggg cacagaggct ccatgtcttc ataattgaag
      841 acgaattcta gataatgaat tctgaaacat aaaatgaatg caattgttgt tgttaacttg
      901 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      961 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     1021 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1081 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     1141 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     1201 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     1261 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     1321 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1381 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1441 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca gtacttaaag gaagagtcct
     1501 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     1561 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1621 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1681 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     1741 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     1801 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     1861 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     1921 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     1981 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     2041 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2101 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2161 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2221 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     2281 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     2341 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     2401 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     2461 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     2521 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     2581 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     2641 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     2701 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     2761 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     2821 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     2881 ggtcgtttgt tcataaacgc ggttcgttcc actgagcgtc agaccccgta gaaaagatca
     2941 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3001 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3061 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3121 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3181 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3241 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3301 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3361 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3421 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3481 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3541 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3601 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     3661 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccggctagcc
     3721 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     3781 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     3841 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     3901 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     3961 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4021 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     4081 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     4141 gactttccaa aatgtcgtaa
//
