LOCUS       sequence-254630-        7762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (mNeonGreen)4-tDeg fluorogenic protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7762)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR
  TITLE     Live imaging of mRNA using RNA-stabilized fluorogenic proteins.
  JOURNAL   Nat Methods. 2019 Sep;16(9):862-865. doi: 10.1038/s41592-019-0531-7.
            Epub 2019 Aug 30.
  PUBMED    31471614
REFERENCE   2  (bases 1 to 7762)
  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: 
            "10.1038/s41592-019-0531-7"; journalName: "Nat Methods"; date: 
            "2019-09"; volume: "16"; issue: "9"; pages: "862-865"
COMMENT     Sequence Label: miniCMV-(mNeonGreen)4-tDeg
FEATURES             Location/Qualifiers
     source          1..7762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..39
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      46..55
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             52..759
                     /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      307..316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2302..3006
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      2554..2563
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3119..3136)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3125..3349
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3395..3823
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3482..3501)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3692..3713
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3832..3852)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3837..4166
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4017..4152
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4079..4098
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4233..5027
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4287..4306)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4897..4916
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5201..5322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5238..5257)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5292..5311
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5371..5387)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5371..5387)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5384..5406)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5395..5411
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5419..5425,5426..5443,5444..5449))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5464..5485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5602..5619)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5773..6361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5853..5872)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6532..7323,7324..7392))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7155..7174
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7393..7497)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7572..7591)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggtaggcgtg tacggtggga ggcctatata agcagagcta agcttgccac catggtgagc
       61 aagggcgagg aggataacat ggcctctctc ccagcgacac atgagttaca catctttggc
      121 tccatcaacg gtgtggactt tgacatggtg ggtcagggca ccggcaatcc aaatgatggt
      181 tatgaggagt taaacctgaa gtccaccaag ggtgacctcc agttctcccc ctggattctg
      241 gtccctcata tcgggtatgg cttccatcag tacctgccct accctgacgg gatgtcgcct
      301 ttccaggccg ccatggtaga tggctccgga taccaagtcc atcgcacaat gcagtttgaa
      361 gatggtgcct cccttactgt taactaccgc tacacctacg agggaagcca catcaaagga
      421 gaggcccagg tgaaggggac tggtttccct gctgacggtc ctgtgatgac caactcgctg
      481 accgctgcgg actggtgcag gtcgaagaag acttacccca acgacaaaac catcatcagt
      541 acctttaagt ggagttacac cactggaaat ggcaagcgct accggagcac tgcgcggacc
      601 acctacacct ttgccaagcc aatggcggct aactatctga agaaccagcc gatgtacgtg
      661 ttccgtaaga cggagctcaa gcactccaag accgagctca acttcaagga gtggcaaaag
      721 gcctttaccg atgtgatggg catggacgag ctgtacaagg gtggacatat gggcacaggg
      781 tccacaggcg gtaccggcgg agtttccaaa ggagaagaag acaatatggc atcactcccc
      841 gcaacccacg agttgcatat tttcggttca attaatggag tagatttcga tatggttggc
      901 cagggaacag gaaacccaaa cgacggatat gaagagctta atctcaaaag taccaaaggc
      961 gatctgcaat tttctccgtg gatactcgtg ccacacattg gatacggatt tcaccaatat
     1021 ctcccgtatc cggatggaat gtcccccttt caagcagcaa tggtggacgg gagtggttat
     1081 caggtacaca gaaccatgca gttcgaggac ggggcttctc tgaccgtaaa ttataggtat
     1141 acttatgaag gctcacatat taagggcgaa gcacaggtta aaggaaccgg gtttcctgcg
     1201 gatggccccg tcatgactaa ttctctgaca gccgcagatt ggtgtcgctc caaaaagaca
     1261 tacccgaatg ataagactat aatctcaaca ttcaaatggt cctatacgac aggcaacggg
     1321 aaacgatata gatccacggc tcgaacaact tacacattcg ctaaacctat ggccgccaat
     1381 tacctcaaaa atcagcccat gtatgtgttt aggaaaaccg aattgaagca ttctaaaacg
     1441 gaacttaatt ttaaggaatg gcagaaggct ttcacagacg taatggggat ggatgaactc
     1501 tataaatcag gtctcgagtc ctcaggggga acgggtgggt ccggaggagt tagtaaaggt
     1561 gaagaggaca atatggcaag tttgcctgcg actcacgagc ttcatatctt tgggtctata
     1621 aatggcgttg acttcgatat ggttggccaa ggtactggca accccaatga cggttacgag
     1681 gagttgaatc tcaagtccac aaaaggtgat cttcagttca gcccttggat tctcgtacct
     1741 catattggat atggctttca ccagtacctt ccatacccag acggtatgtc accctttcaa
     1801 gctgcgatgg tggatggttc cggctatcag gtccaccgaa cgatgcaatt cgaggacggg
     1861 gccagcctca ccgttaatta taggtacacc tatgagggaa gtcacataaa gggagaagcc
     1921 caagtgaaag gaacaggatt cccagctgat ggtccagtaa tgacgaactc cttgacagcg
     1981 gctgactggt gtagaagcaa aaagacgtat cctaatgaca agaccatcat tagcactttc
     2041 aaatggagtt ataccacagg aaacggcaaa cggtacagaa gcactgctag aactacctac
     2101 actttcgcaa agccgatggc tgcaaactat ttgaagaatc agcccatgta cgtgtttcga
     2161 aaaacggaac ttaagcacag taagactgaa cttaatttca aggagtggca gaaggcgttc
     2221 acggatgtca tgggtatgga tgaactgtat aagggagggt ctggcactgg gggcactgcc
     2281 agcagcggat ccggtggcgg tgtgagcaag ggcgaggagg ataacatggc ctctctccca
     2341 gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt
     2401 cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt
     2461 gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac
     2521 ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggatac
     2581 caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac
     2641 acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct
     2701 gacggtcctg tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaagact
     2761 taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc
     2821 aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac
     2881 tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc
     2941 gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg
     3001 tacaagggcg gaagatccgg tggtggttct ggtcctcgtc cccgtggtac tcgtggtaaa
     3061 ggtcgccgta ttcgtcgccg cggttaatct agagggcccg tttaaacccg ctgatcagcc
     3121 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3181 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3241 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3301 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3361 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3421 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3481 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3541 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3601 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3661 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3721 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3781 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3841 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3901 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3961 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4021 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4081 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4141 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4201 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4261 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4321 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4381 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4441 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4501 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4561 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4621 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4681 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4741 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4801 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4861 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4921 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4981 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5041 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5101 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5161 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5221 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5281 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5341 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5401 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5461 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5521 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5581 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5641 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5701 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5761 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5821 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5881 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5941 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6001 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6061 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6121 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6181 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6241 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6301 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6361 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6421 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6481 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6541 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6601 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6661 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6721 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6781 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6841 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6901 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6961 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7021 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7081 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7141 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7201 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7261 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7321 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7381 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7441 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7501 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7561 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     7621 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     7681 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     7741 gtacgggcca gatatacgcg tt
//
