LOCUS       sequence-254682-        8122 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 8122)
  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 8122)
  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: UbC-(mNeonGreen)4-tDeg
FEATURES             Location/Qualifiers
     source          1..8122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        235..633
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     regulatory      640..649
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             646..1353
                     /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      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2896..3600
                     /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      3148..3157
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3713..3730)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3719..3943
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3989..4417
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4076..4095)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4286..4307
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4426..4446)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4431..4760
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4611..4746
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4673..4692
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4827..5621
                     /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(4881..4900)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5491..5510
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5795..5916
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5832..5851)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5886..5905
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5965..5981)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5965..5981)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5978..6000)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5989..6005
                     /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(6013..6019,6020..6037,6038..6043))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6058..6079
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6196..6213)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6367..6955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6447..6466)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7126..7917,7918..7986))
                     /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     7749..7768
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7987..8091)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttggcctc
      241 cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt
      301 cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc
      361 gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg
      421 acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt
      481 agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat
      541 ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt
      601 tcttgtttgt ggatcgctgt gatcgtcact tggaagcttg ccaccatggt gagcaagggc
      661 gaggaggata acatggcctc tctcccagcg acacatgagt tacacatctt tggctccatc
      721 aacggtgtgg actttgacat ggtgggtcag ggcaccggca atccaaatga tggttatgag
      781 gagttaaacc tgaagtccac caagggtgac ctccagttct ccccctggat tctggtccct
      841 catatcgggt atggcttcca tcagtacctg ccctaccctg acgggatgtc gcctttccag
      901 gccgccatgg tagatggctc cggataccaa gtccatcgca caatgcagtt tgaagatggt
      961 gcctccctta ctgttaacta ccgctacacc tacgagggaa gccacatcaa aggagaggcc
     1021 caggtgaagg ggactggttt ccctgctgac ggtcctgtga tgaccaactc gctgaccgct
     1081 gcggactggt gcaggtcgaa gaagacttac cccaacgaca aaaccatcat cagtaccttt
     1141 aagtggagtt acaccactgg aaatggcaag cgctaccgga gcactgcgcg gaccacctac
     1201 acctttgcca agccaatggc ggctaactat ctgaagaacc agccgatgta cgtgttccgt
     1261 aagacggagc tcaagcactc caagaccgag ctcaacttca aggagtggca aaaggccttt
     1321 accgatgtga tgggcatgga cgagctgtac aagggtggac atatgggcac agggtccaca
     1381 ggcggtaccg gcggagtttc caaaggagaa gaagacaata tggcatcact ccccgcaacc
     1441 cacgagttgc atattttcgg ttcaattaat ggagtagatt tcgatatggt tggccaggga
     1501 acaggaaacc caaacgacgg atatgaagag cttaatctca aaagtaccaa aggcgatctg
     1561 caattttctc cgtggatact cgtgccacac attggatacg gatttcacca atatctcccg
     1621 tatccggatg gaatgtcccc ctttcaagca gcaatggtgg acgggagtgg ttatcaggta
     1681 cacagaacca tgcagttcga ggacggggct tctctgaccg taaattatag gtatacttat
     1741 gaaggctcac atattaaggg cgaagcacag gttaaaggaa ccgggtttcc tgcggatggc
     1801 cccgtcatga ctaattctct gacagccgca gattggtgtc gctccaaaaa gacatacccg
     1861 aatgataaga ctataatctc aacattcaaa tggtcctata cgacaggcaa cgggaaacga
     1921 tatagatcca cggctcgaac aacttacaca ttcgctaaac ctatggccgc caattacctc
     1981 aaaaatcagc ccatgtatgt gtttaggaaa accgaattga agcattctaa aacggaactt
     2041 aattttaagg aatggcagaa ggctttcaca gacgtaatgg ggatggatga actctataaa
     2101 tcaggtctcg agtcctcagg gggaacgggt gggtccggag gagttagtaa aggtgaagag
     2161 gacaatatgg caagtttgcc tgcgactcac gagcttcata tctttgggtc tataaatggc
     2221 gttgacttcg atatggttgg ccaaggtact ggcaacccca atgacggtta cgaggagttg
     2281 aatctcaagt ccacaaaagg tgatcttcag ttcagccctt ggattctcgt acctcatatt
     2341 ggatatggct ttcaccagta ccttccatac ccagacggta tgtcaccctt tcaagctgcg
     2401 atggtggatg gttccggcta tcaggtccac cgaacgatgc aattcgagga cggggccagc
     2461 ctcaccgtta attataggta cacctatgag ggaagtcaca taaagggaga agcccaagtg
     2521 aaaggaacag gattcccagc tgatggtcca gtaatgacga actccttgac agcggctgac
     2581 tggtgtagaa gcaaaaagac gtatcctaat gacaagacca tcattagcac tttcaaatgg
     2641 agttatacca caggaaacgg caaacggtac agaagcactg ctagaactac ctacactttc
     2701 gcaaagccga tggctgcaaa ctatttgaag aatcagccca tgtacgtgtt tcgaaaaacg
     2761 gaacttaagc acagtaagac tgaacttaat ttcaaggagt ggcagaaggc gttcacggat
     2821 gtcatgggta tggatgaact gtataaggga gggtctggca ctgggggcac tgccagcagc
     2881 ggatccggtg gcggtgtgag caagggcgag gaggataaca tggcctctct cccagcgaca
     2941 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
     3001 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
     3061 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
     3121 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ataccaagtc
     3181 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
     3241 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     3301 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     3361 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     3421 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     3481 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     3541 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     3601 ggcggaagat ccggtggtgg ttctggtcct cgtccccgtg gtactcgtgg taaaggtcgc
     3661 cgtattcgtc gccgcggtta atctagaggg cccgtttaaa cccgctgatc agcctcgact
     3721 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3781 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3841 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3901 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3961 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4021 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4081 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4141 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4201 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4261 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4321 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4381 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4441 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4501 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4561 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4621 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4681 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4741 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4801 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4861 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4921 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4981 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5041 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5101 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5161 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5221 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5281 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5341 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5401 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5461 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5521 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5581 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5641 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5701 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5761 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5821 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5881 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5941 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6001 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6061 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6121 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6181 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6241 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6301 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6361 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6421 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6481 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6541 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6601 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6661 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6721 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6781 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6841 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6901 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6961 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7021 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7081 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7141 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7201 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7261 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7321 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7381 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7441 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7501 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7561 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7621 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7681 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7741 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7801 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7861 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7921 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7981 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8041 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8101 ccgaaaagtg ccacctgacg tc
//
