LOCUS       sequence-452467-        4175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mPapaya, for MuSIC barcode construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4175)
  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 4175)
  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-mPapaya
FEATURES             Location/Qualifiers
     source          1..4175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      458..467
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(515..718)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(544..564)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        719..1022
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1203..1791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1283..1302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1850..1897
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1922..1941
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2129..2923)
                     /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(2129..2923)
                     /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(2240..2259)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2850..2869
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2958..3154)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(2966..3176)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2972..3107
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3200..3220
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3264..3719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3351..3370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3561..3582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3726..3847
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3737..3756)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3791..3810
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3926..4175,1..461))
                     /codon_start=1
                     /product="monomeric yellow fluorescent protein derived from
                     the Zoanthus fluorescent protein zFP538"
                     /label=mPapaya1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEGQSKHGLKEEMTVKYHMEGCVNGHKFVITGEGIGNPFKG
                     KQTANLCVIEGGPLPFSEDILSPGFKYGDRIFTEYPQDIVDYFKNSCPAGYTWERSYLF
                     EDGAVCRCNVDITVSEKENCIYHKSIFRGVNFPADGPVMKKMTTNWEASTEKIVPVPKQ
                     GILKGKVKMCLLLKDGGRYHCQFDTVYKAKSVPSKMPEWHFIQHKLLREDRSDAKNQKW
                     QLTEHAIAGMDELYK"
ORIGIN
        1 gtactagctt cccaattagt ggtcattttc ttcattacgg ggccgtcggc gggaaaattc
       61 acccctctga agatactttt gtgatagatg cagttctcct tttcagagac tgttatgtcc
      121 acgttgcatc gacagaccgc tccgtcctca aagaggtagc tcctttccca cgtatagccc
      181 gctggacatg agttcttgaa gtaatctaca atatcctgtg ggtactctgt gaaaatccgg
      241 tcaccatatt taaaacccgg acttaaaatg tcctccgaga acggcagcgg gcctccttct
      301 atcacacaca aatttgcggt ttgcttaccc ttaaaagggt ttccaatgcc ctctccagta
      361 atgacaaatt tgtgaccatt aacgcaacct tccatgtggt acttcaccgt catctcctcc
      421 tttagtccat gcttcgattg cccctcgccc ttgctcacca tggtggcggc ggatccattt
      481 cgataagcca gtaagcacga ggctgatcag cggtagctct gcttatatag acctcccacc
      541 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
      601 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
      661 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
      721 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
      781 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
      841 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
      901 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
      961 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     1021 cggctagccg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     1081 aataactaat gcatggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1141 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1201 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1261 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1321 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1381 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1441 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1501 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1561 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1621 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     1681 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1741 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1801 ttgatctttt ctacggggtc tgacgctcag tggaacgaac cgcgtttatg aacaaacgac
     1861 ccaacaccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt
     1921 attgtctcct tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg
     1981 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     2041 ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg
     2101 tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg
     2161 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
     2221 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
     2281 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
     2341 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
     2401 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
     2461 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
     2521 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
     2581 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
     2641 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
     2701 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
     2761 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
     2821 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
     2881 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
     2941 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     3001 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     3061 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     3121 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     3181 gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct
     3241 caggactctt cctttaagta ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3301 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3361 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3421 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3481 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3541 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3601 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3661 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3721 cgccttaaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     3781 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     3841 acaagttaac aacaacaatt gcattcattt tatgtttcag aattcattat ctagaattcg
     3901 tcttcaatta tgaagacatg gagcccttgt acagctcgtc catgcctgcg attgcatgtt
     3961 ctgttaattg ccatttctgg tttttagcat cagagcggtc ctccctaagg agcttatgct
     4021 gtataaaatg ccattctggc atcttggagg gcactgattt agctttatat accgtatcaa
     4081 actggcaatg ataacgaccg ccatccttca gcaacaggca cattttgacc tttcccttta
     4141 atatcccttg ctttggaaca ggcacaattt tctcg
//
