LOCUS       sequence-452455-        4181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mCerulean3, for MuSIC barcode construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4181)
  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 4181)
  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-mCerulean3
FEATURES             Location/Qualifiers
     source          1..4181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(76..870)
                     /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(76..870)
                     /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(187..206)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     797..816
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(905..1101)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(913..1123)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      919..1054
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1147..1167
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1211..1666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1298..1317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1508..1529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1673..1794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1684..1703)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1738..1757
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1873..2583,2584..2586,2587..2589))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1915..1936)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2264..2283
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2523..2544
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2586..2595
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2643..2846)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2672..2692)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2847..3150
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3331..3919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3411..3430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3978..4025
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4050..4069
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga
       61 accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga
      121 atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc
      181 ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg
      241 gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc
      301 atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa
      361 cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc
      421 ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca
      481 ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc
      541 ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca
      601 gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc
      661 cagccacgat agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt
      721 cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca
      781 gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga
      841 acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc
      901 gatctttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag
      961 aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     1021 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg
     1081 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     1141 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
     1201 ttaagtactg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1261 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1321 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1381 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1441 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1501 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     1561 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     1621 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacgc cttaagatac
     1681 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1741 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1801 aacaattgca ttcattttat gtttcagaat tcttactact gaattcgtct tcaattatga
     1861 agacatggag cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     1921 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagct tggactgggt
     1981 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     2041 gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg cagttgaggc cgaagttggc
     2101 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgccgtgga tggcgttgta
     2161 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     2221 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     2281 gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa
     2341 gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac tgcacgcccc agctcagggt
     2401 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     2461 cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt
     2521 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     2581 gctcaccatg gtggcggcgg atccatttcg ataagccagt aagcacgagg ctgatcagcg
     2641 gtagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     2701 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     2761 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     2821 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     2881 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     2941 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     3001 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     3061 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     3121 ggcgggccat ttaccgtaag ttatgtaacg gctagccgga actccatata tgggctatga
     3181 actaatgacc ccgtaattga ttactattaa taactaatgc atggcggtaa tacggttatc
     3241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3961 gaacgaaccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat
     4021 tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc
     4081 cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc
     4141 gtcccggaaa acgattccga agcccaacct ttcatagaag g
//
