LOCUS       sequence-452468-        4181 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of mVenus, 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 F, Biswas T, 
            Ashy KC, Woods SK, Livingston MWT, Blenner MA, Birtwistle MR
  TITLE     Genetically-Encoded Fluorescence Barcodes for Single-Cell Analysis.
  JOURNAL   bioRxiv [Preprint]. 2024 Oct 24:2024.10.23.619855. doi: 
            10.1101/2024.10.23.619855.
  PUBMED    39484616
REFERENCE   2  (bases 1 to 4181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Oct 24:2024.10.23.619855. doi: 
            10.1101/2024.10.23.619855."
COMMENT     Sequence Label: pR-mVenus
FEATURES             Location/Qualifiers
     source          1..4181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(113..907)
                     /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(113..907)
                     /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(224..243)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     834..853
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(942..1138)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(950..1160)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      956..1091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1184..1204
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1248..1703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1335..1354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1545..1566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1710..1831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1721..1740)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1775..1794
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1910..2620,2621..2623,2624..2626))
                     /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(1952..1973)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2301..2320
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2560..2581
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2623..2632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2680..2883)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2709..2729)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2884..3187
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3368..3956)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3448..3467)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4015..4062
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4087..4106
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
ORIGIN
        1 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
       61 gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc gctcagaaga
      121 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
      181 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
      241 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
      301 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
      361 cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
      421 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
      481 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
      541 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
      601 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
      661 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
      721 cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
      781 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
      841 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
      901 caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa gcctaggcct
      961 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
     1021 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
     1081 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
     1141 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     1201 ttccacagct ggttctttcc gcctcaggac tcttccttta agtactgacg cgccctgtag
     1261 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1321 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1381 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1441 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1501 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1561 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1621 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     1681 caaaatatta acgcttacaa tttacgcctt aagatacatt gatgagtttg gacaaaccac
     1741 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1801 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1861 tcagaattcg gcaactcgaa ttcgtcttca attatgaaga catggagccc ttgtagagct
     1921 cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc
     1981 gcttctcgtt ggggtctttg ctcagcttgg actggtagct caggtagtgg ttgtcgggca
     2041 gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc
     2101 cgccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc ttctgcttgt
     2161 cggcggtgat atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt
     2221 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     2281 cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct
     2341 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     2401 ggtagcgggc gaagcactgc aggccgtagc ccagggtggt cacgagggtg ggccagggca
     2461 cgggcagctt gccggtggtg cagatcagct tcagggtcag cttgccgtag gtggcatcgc
     2521 cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     2581 ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg gcggcggatc
     2641 catttcgata agccagtaag cacgaggctg atcagcggta gctctgctta tatagacctc
     2701 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
     2761 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
     2821 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     2881 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     2941 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
     3001 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
     3061 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
     3121 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
     3181 tgtaacggct agccggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     3241 ctattaataa ctaatgcatg gcggtaatac ggttatccac agaatcaggg gataacgcag
     3301 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3361 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3421 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3481 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3541 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3601 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3661 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3721 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3781 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3841 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3901 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3961 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaccgcgt ttatgaacaa
     4021 acgacccaac accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt
     4081 ccggtattgt ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca
     4141 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc c
//
