LOCUS       sequence-452483-        3761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mBeRFP and extra insert, for MuSIC barcode 
            construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3761)
  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 3761)
  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-mBeRFP
FEATURES             Location/Qualifiers
     source          1..3761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    325..446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(453..908)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(590..611)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     802..821
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        934..1036
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1037..1696
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1103..1122)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     polyA_signal    1737..1858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1912..2500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2401..2420
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2675..2978
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2979..3182
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3133..3153
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3209..3265
                     /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"
     CDS             3275..3328
                     /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"
     regulatory      3338..3347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aaacactcgg ttgggaagcc tccacagaga tgctgtaccc agctgacgga ggtctggaag
       61 gaagagacta catggctctg aagctcgtgg gcgggggcca cctgatctgc aacgccaaga
      121 ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg
      181 acaggagact ggaaagaatc aaggaggccg acaaagagac aagcgtcgag cagcacgagg
      241 tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaataa gagttgccat
      301 tcatgtcttc tgaaacataa aatgaacttg tttattgcag cttataatgg ttacaaataa
      361 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
      421 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
      481 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
      541 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
      601 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
      661 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
      721 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
      781 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
      841 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
      901 gggcgcgtca gtacttaacg gaagatcact tcgtgatcgg cacgtaagag gttccaactt
      961 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     1021 agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca
     1081 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
     1141 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
     1201 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat
     1261 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
     1321 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
     1381 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
     1441 taaagggttt attgagaata tgtttttcgt atcagccaat ccctgggtga gtttcaccag
     1501 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     1561 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     1621 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     1681 gcagggcggg gcgtaatcaa acatgagaat tggtcgcgcg agcaattcag acatgataag
     1741 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1801 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttgg
     1861 catttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1921 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1981 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2041 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2101 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2161 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2221 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2281 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2341 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2401 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2461 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2521 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2581 tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg
     2641 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     2701 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     2761 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     2821 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2881 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2941 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     3001 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     3061 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     3121 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     3181 ctggtttagt gaagaagacg gctccggcgc cacaaacttc tctctgctaa agcaagcagg
     3241 tgatgttgaa gaaaaccccg ggcctggcag cggcgagggc aggggaagtc tacttacatg
     3301 cggggacgtg gaggaaaatc ccggcccagg atccgccgcc accatggtgt ctaagggcga
     3361 agagctgatt aaggagaaca tgcatatgaa gctgtacatg gagggcaccg tgaacaacca
     3421 ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     3481 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt
     3541 catgtacggc agcaaaacct tcatcaacca cacccagggc atccccgact tctttaagca
     3601 gtccttccct gagggcttca catgggagag atccaccaca tacgaggacg ggggcgtgct
     3661 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
     3721 aggggtgaac ttcccatcca acggccctgt gatgcagaag a
//
