LOCUS       sequence-390516-        4722 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 4722)
  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 4722)
  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: NeonGreen
FEATURES             Location/Qualifiers
     source          1..4722
                     /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..1431
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      979..988
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1432..1497
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1621..1742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1658..1677)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1712..1731
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1749..2204)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1886..1907)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2098..2117
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2231..2335
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2337..2694
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2337..2528
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2360..2380)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2545..2680
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2729..3523
                     /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             2729..3523
                     /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(2783..2802)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3393..3412
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3711..3730)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3755..3802
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4131..4719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4620..4639
                     /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 ggaggataac atggcctctc tcccagcgac acatgagtta
      781 cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg caccggcaat
      841 ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct ccagttctcc
      901 ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc ctaccctgac
      961 gggatgtcgc ctttccaggc cgccatggta gatggctccg gataccaagt ccatcgcaca
     1021 atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta cgagggaagc
     1081 cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg tcctgtgatg
     1141 accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc caacgacaaa
     1201 accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg ctaccggagc
     1261 actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct gaagaaccag
     1321 ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct caacttcaag
     1381 gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa gtccggactc
     1441 agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc
     1501 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1561 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1621 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     1681 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     1741 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     1801 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     1861 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1921 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1981 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2041 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2101 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2161 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     2221 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2281 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     2341 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2401 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2461 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2521 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2581 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2641 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     2701 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     2761 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     2821 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     2881 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     2941 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3001 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3061 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3121 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3181 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3241 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3301 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3361 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3421 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3481 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3541 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3601 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3661 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     3721 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     3781 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     3841 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     3901 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     3961 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4021 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4081 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4141 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4201 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4261 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4321 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4381 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4441 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4501 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4561 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4621 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4681 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa ac
//
