LOCUS       sequence-414192-        4731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Monomer expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4731)
  AUTHORS   Petrich A, Aji AK, Dunsing V, Chiantia S
  TITLE     Benchmarking of novel green fluorescent proteins for the 
            quantification of protein oligomerization in living cells.
  JOURNAL   PLoS One. 2023 Aug 3;18(8):e0285486. doi: 
            10.1371/journal.pone.0285486. eCollection 2023.
  PUBMED    37535571
REFERENCE   2  (bases 1 to 4731)
  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"; journalName: "PLoS One.";
            date: "2023-08-3"; volume: "18(8):e0285486. doi"; pages: " 
            10.1371/journal.pone.0285486. eCollection 2023"
COMMENT     Sequence Label: Gamillus
FEATURES             Location/Qualifiers
     source          1..4731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        172..475
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        476..679
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     630..650
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      718..727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             724..1440
                     /codon_start=1
                     /product="acid-tolerant monomeric green fluorescent protein
                     (Shinoda et al., 2018)"
                     /label=Gamillus
                     /translation="MVSKGEEASGRALFQYPMTSKIELNGEINGKKFKVAGEGFTPSSG
                     RFNMHAYCTTGDLPMSWVVIASPLQYGFHMFAHYPEDITHFFQECFPGSYTLDRTLRME
                     GDGTLTTHHEYSLEDGCVTSKTTLNASGFDPKGATMTKSFVKQLPNEVKITPHGPNGIR
                     LTSTVLYLKEDGTIQIGTQDCIVTPVGGRKVTQPKAHFLHTQIIQKKDPNDTRDHIVQT
                     ELAVAGNLWHGMDELYK"
     misc_feature    1441..1506
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1630..1751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1667..1686)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1721..1740
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1758..2213)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1895..1916)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2107..2126
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2240..2344
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2346..2703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2346..2537
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2369..2389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2554..2689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2738..3532
                     /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             2738..3532
                     /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(2792..2811)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3402..3421
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3720..3739)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3764..3811
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4140..4728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4629..4648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
       61 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
      121 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      181 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      241 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      301 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      361 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      421 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      481 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      541 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      601 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      661 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc
      721 accatggtga gcaagggcga ggaggcatct ggcagagccc tgttccagta ccccatgacc
      781 agcaagatcg agctgaacgg cgagatcaac ggcaagaaat tcaaggtggc cggcgagggc
      841 ttcaccccca gcagcggcag attcaacatg cacgcctact gcaccaccgg cgacctgcct
      901 atgagctggg tcgtgattgc cagccccctc cagtacggct tccacatgtt cgcccactac
      961 cccgaggaca tcacacactt tttccaggaa tgcttccccg gcagctacac cctggaccgg
     1021 accctgagaa tggaaggcga cggcaccctg accacccacc acgagtacag cctggaggac
     1081 ggctgcgtga cctccaagac caccctgaat gccagcggct tcgaccctaa gggcgccacc
     1141 atgaccaaga gcttcgtgaa acaactgcct aacgaggtga agatcacccc ccacggcccc
     1201 aacggcatca gactgaccag caccgtgctg tacctgaagg aggatggcac catccagatc
     1261 ggcacccagg actgcatcgt gacccctgtg ggcggaagga aagtgaccca gcccaaggcc
     1321 cacttcctgc acacccagat catccagaag aaggacccca acgacacccg ggaccacatc
     1381 gtgcagacag aactggccgt ggccggcaat ctgtggcacg gcatggacga gctgtacaag
     1441 tccggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg
     1501 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     1561 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     1621 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     1681 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     1741 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     1801 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     1861 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     1921 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     1981 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2041 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2101 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2161 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     2221 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2281 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2341 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2401 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2461 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2521 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2581 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2641 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2701 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     2761 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     2821 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     2881 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     2941 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3001 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3061 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3121 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3181 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3241 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     3301 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3361 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3421 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     3481 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     3541 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     3601 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     3661 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     3721 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3781 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     3841 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     3901 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     3961 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     4021 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4081 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4261 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa c
//
