LOCUS       sequence-452448-        4172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mTFP1, 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-mTFP1
FEATURES             Location/Qualifiers
     source          1..4172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     398..417
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(716..735)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    760..807
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      866..1454
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1355..1374
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1635..1938
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1939..2142
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2093..2113
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2190..2199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2196..2903
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    2982..3103
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3019..3038)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3073..3092
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3110..3565)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3247..3268)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3459..3478
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3609..3629)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3653..3863
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3675..3871
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3722..3857
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(3906..4172,1..528)
                     /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             join(3906..4172,1..528)
                     /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(3960..3979)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
       61 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      121 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
      181 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
      241 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
      301 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
      361 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
      421 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
      481 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
      541 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
      601 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
      661 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
      721 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
      781 aaacgcacgg tgttgggtcg tttgttcata aacgcggttc gttccactga gcgtcagacc
      841 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      901 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      961 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1021 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1081 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1141 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1201 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1261 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1321 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1381 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1441 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     1501 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     1561 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     1621 gagttccggc tagccgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1681 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1741 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1801 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1861 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1921 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     1981 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     2041 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     2101 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctaccgctga tcagcctcgt
     2161 gcttactggc ttatcgaaat ggatccgccg ccaccatggt gagcaagggc gaggagacta
     2221 caatgggcgt aatcaagccc gacatgaaga tcaagctgaa gatggagggc aacgtgaatg
     2281 gccacgcctt cgtgatcgag ggcgagggcg agggcaagcc ctacgacggc accaacacca
     2341 tcaacctgga ggtgaaggag ggagcccccc tgcccttctc ctacgacatt ctgaccaccg
     2401 cgttcgccta cggcaacagg gccttcacca agtaccccga cgacatcccc aactacttca
     2461 agcagtcctt ccccgagggc tactcttggg agcgcaccat gaccttcgag gacaagggca
     2521 tcgtgaaggt gaagtccgac atctccatgg aggaggactc cttcatctac gagatacacc
     2581 tcaagggcga gaacttcccc cccaacggcc ccgtgatgca gaagaaaacc accggctggg
     2641 acgcctccac cgagaggatg tacgtgcgcg acggcgtgct gaagggcgac gtcaagcaca
     2701 agctgctgct ggagggcggc ggccaccacc gcgttgactt caagaccatc tacagggcca
     2761 agaaggcggt gaagctgccc gactatcact ttgtggacca ccgcatcgag atcctgaacc
     2821 acgacaagga ctacaacaag gtgaccgttt acgagagcgc cgtggcccgc aactccaccg
     2881 acggcatgga cgagctgtac aagggctcca tgtcttcata attgaagacg aattcagctt
     2941 cttgaattct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3001 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3061 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3121 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3181 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3241 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3301 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3361 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3421 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3481 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3541 cttaatgcgc cgctacaggg cgcgtcagta cttaaaggaa gagtcctgag gcggaaagaa
     3601 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3661 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3721 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3781 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     3841 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     3901 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3961 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4021 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4081 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4141 ctgtgctcga cgttgtcact gaagcgggaa gg
//
