LOCUS       sequence-470403-        3767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mPapaya and extra insert, for MuSIC barcode 
            construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3767)
  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 3767)
  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: pM-mPapaya
FEATURES             Location/Qualifiers
     source          1..3767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      417..426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(436..489)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(499..555)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     promoter        complement(582..785)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(611..631)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        786..1089
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1264..1852)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1344..1363)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1906..2027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2068..2727)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2642..2661
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2728..2830)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2856..3311
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2943..2962)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3153..3174
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3318..3439
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(3474..3767,1..420))
                     /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 gccgtcggcg ggaaaattca cccctctgaa gatacttttg tgatagatgc agttctcctt
       61 ttcagagact gttatgtcca cgttgcatcg acagaccgct ccgtcctcaa agaggtagct
      121 cctttcccac gtatagcccg ctggacatga gttcttgaag taatctacaa tatcctgtgg
      181 gtactctgtg aaaatccggt caccatattt aaaacccgga cttaaaatgt cctccgagaa
      241 cggcagcggg cctccttcta tcacacacaa atttgcggtt tgcttaccct taaaagggtt
      301 tccaatgccc tctccagtaa tgacaaattt gtgaccatta acgcaacctt ccatgtggta
      361 cttcaccgtc atctcctcct ttagtccatg cttcgattgc ccctcgccct tgctcaccat
      421 ggtggcggcg gatcctgggc cgggattttc ctccacgtcc ccgcatgtaa gtagacttcc
      481 cctgccctcg ccgctgccag gcccggggtt ttcttcaaca tcacctgctt gctttagcag
      541 agagaagttt gtggcgccgg agccgtcttc ttcactaaac cagctctgct tatatagacc
      601 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
      661 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
      721 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
      781 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
      841 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
      901 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
      961 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     1021 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     1081 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     1141 taataactaa tgcatggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1201 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1261 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1321 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1381 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1441 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1501 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1561 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1621 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1681 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1741 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1801 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1861 tttgatcttt tctacggggt ctgacgctca gtggaacgaa atgccaactt gtttattgca
     1921 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     1981 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgaatt
     2041 gctcgcgcga ccaattctca tgtttgatta cgccccgccc tgccactcat cgcagtactg
     2101 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     2161 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     2221 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     2281 gggattggct gatacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     2341 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     2401 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     2461 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     2521 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     2581 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     2641 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     2701 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     2761 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     2821 gtgccgatca cgaagtgatc ttccgttaag tactgacgcg ccctgtagcg gcgcattaag
     2881 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2941 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3001 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3061 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3121 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3181 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3241 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3301 gcttacaatt tacgccttaa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3361 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3421 aagctgcaat aaacaagttc attttatgtt tcagaagaca tgaatggcaa ctcttacttg
     3481 tacagctcgt ccatgcctgc gattgcatgt tctgttaatt gccatttctg gtttttagca
     3541 tcagagcggt cctccctaag gagcttatgc tgtataaaat gccattctgg catcttggag
     3601 ggcactgatt tagctttata taccgtatca aactggcaat gataacgacc gccatccttc
     3661 agcaacaggc acattttgac ctttcccttt aatatccctt gctttggaac aggcacaatt
     3721 ttctcggtac tagcttccca attagtggtc attttcttca ttacggg
//
