LOCUS       sequence-452496-        3079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of a single fluorescent protein and extra 
            insert, for MuSIC barcode construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3079)
  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 3079)
  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: pMulti
FEATURES             Location/Qualifiers
     source          1..3079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    414..535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      589..1177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1078..1097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1352..1655
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1656..1859
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1810..1830
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1886..1942
                     /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             1952..2005
                     /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"
     polyA_signal    2081..2202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2209..2664)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2346..2367)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2558..2577
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2690..2792
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2793..3079,1..373)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2859..2878)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
       61 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
      121 agggtttatt gagaatatgt ttttcgtatc agccaatccc tgggtgagtt tcaccagttt
      181 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
      241 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
      301 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
      361 gggcggggcg taatcaaaca tgagaattgg tcgcgcgagc aattcagaca tgataagata
      421 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      481 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttggcat
      541 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      601 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      661 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      721 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1141 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1201 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1261 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     1321 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     1381 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     1441 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     1501 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     1561 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1621 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     1681 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     1741 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     1801 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     1861 gtttagtgaa gaagacggct ccggcgccac aaacttctct ctgctaaagc aagcaggtga
     1921 tgttgaagaa aaccccgggc ctggcagcgg cgagggcagg ggaagtctac ttacatgcgg
     1981 ggacgtggag gaaaatcccg gcccaggatc cgagaccctc tggctaacta gagaacccac
     2041 tgcttactgg tctcattcat gtcttctgaa acataaaatg aacttgttta ttgcagctta
     2101 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2161 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     2221 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     2281 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     2341 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2401 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2461 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2521 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2581 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2641 ttaatgcgcc gctacagggc gcgtcagtac ttaacggaag atcacttcgt gatcggcacg
     2701 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     2761 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     2821 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     2881 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     2941 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     3001 atccggaatt ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     3061 cttgttacac cgttttcca
//
