LOCUS       sequence-452491-        3773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mVenus and extra insert, for MuSIC barcode 
            construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3773)
  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 3773)
  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-mVenus
FEATURES             Location/Qualifiers
     source          1..3773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(56..769,770..772,773..775))
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(101..122)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     450..469
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     709..730
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      772..781
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(791..844)
                     /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(854..910)
                     /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(937..1140)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(966..986)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1141..1444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1619..2207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1699..1718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2261..2382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2423..3082)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2997..3016
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3083..3185)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      3211..3666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3298..3317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3508..3529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    join(3673..3773,1..21)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ataagctgca ataaacaagt tcattttatg tttcagaaga catgaatggc aactcttact
       61 tgtagagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
      121 tgtgatcgcg cttctcgttg gggtctttgc tcagcttgga ctggtagctc aggtagtggt
      181 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
      241 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct
      301 tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
      361 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
      421 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
      481 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
      541 tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc cagggtggtc acgagggtgg
      601 gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc ttgccgtagg
      661 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
      721 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
      781 cggcggatcc tgggccggga ttttcctcca cgtccccgca tgtaagtaga cttcccctgc
      841 cctcgccgct gccaggcccg gggttttctt caacatcacc tgcttgcttt agcagagaga
      901 agtttgtggc gccggagccg tcttcttcac taaaccagct ctgcttatat agacctccca
      961 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     1021 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     1081 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     1141 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     1201 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     1261 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     1321 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     1381 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     1441 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     1501 actaatgcat ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1561 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1621 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1681 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1741 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1801 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1861 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     1921 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     1981 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2041 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2101 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2161 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2221 tcttttctac ggggtctgac gctcagtgga acgaaatgcc aacttgttta ttgcagctta
     2281 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2341 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gaattgctcg
     2401 cgcgaccaat tctcatgttt gattacgccc cgccctgcca ctcatcgcag tactgttgta
     2461 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     2521 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     2581 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2641 tggctgatac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     2701 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2761 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2821 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat
     2881 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     2941 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     3001 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     3061 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     3121 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     3181 gatcacgaag tgatcttccg ttaagtactg acgcgccctg tagcggcgca ttaagcgcgg
     3241 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3301 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3361 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3421 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3481 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3541 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3601 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3661 caatttacgc cttaagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     3721 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc att
//
