LOCUS       sequence-248442-        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"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        235..273
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      280..289
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             286..993
                     /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      541..550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2536..3240
                     /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      2788..2797
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3353..3370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3359..3586
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3632..4060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3719..3738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3929..3950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4069..4089)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4074..4403
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4254..4389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4316..4335
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4470..5264
                     /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(4524..4543)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5134..5153
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5438..5559
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5475..5494)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5529..5548
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5608..5624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5608..5624)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5621..5643)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5632..5648
                     /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(5656..5662,5663..5680,5681..5686))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5701..5722
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5839..5856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6010..6595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6090..6109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6766..7557,7558..7626))
                     /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     7389..7408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7627..7731)
                     /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 cgttggtagg
      241 cgtgtacggt gggaggccta tataagcaga gctaagcttg ccaccatggt gagcaagggc
      301 gaggaggata acatggcctc tctcccagcg acacatgagt tacacatctt tggctccatc
      361 aacggtgtgg actttgacat ggtgggtcag ggcaccggca atccaaatga tggttatgag
      421 gagttaaacc tgaagtccac caagggtgac ctccagttct ccccctggat tctggtccct
      481 catatcgggt atggcttcca tcagtacctg ccctaccctg acgggatgtc gcctttccag
      541 gccgccatgg tagatggctc cggataccaa gtccatcgca caatgcagtt tgaagatggt
      601 gcctccctta ctgttaacta ccgctacacc tacgagggaa gccacatcaa aggagaggcc
      661 caggtgaagg ggactggttt ccctgctgac ggtcctgtga tgaccaactc gctgaccgct
      721 gcggactggt gcaggtcgaa gaagacttac cccaacgaca aaaccatcat cagtaccttt
      781 aagtggagtt acaccactgg aaatggcaag cgctaccgga gcactgcgcg gaccacctac
      841 acctttgcca agccaatggc ggctaactat ctgaagaacc agccgatgta cgtgttccgt
      901 aagacggagc tcaagcactc caagaccgag ctcaacttca aggagtggca aaaggccttt
      961 accgatgtga tgggcatgga cgagctgtac aagggtggac atatgggcac agggtccaca
     1021 ggcggtaccg gcggagtttc caaaggagaa gaagacaata tggcatcact ccccgcaacc
     1081 cacgagttgc atattttcgg ttcaattaat ggagtagatt tcgatatggt tggccaggga
     1141 acaggaaacc caaacgacgg atatgaagag cttaatctca aaagtaccaa aggcgatctg
     1201 caattttctc cgtggatact cgtgccacac attggatacg gatttcacca atatctcccg
     1261 tatccggatg gaatgtcccc ctttcaagca gcaatggtgg acgggagtgg ttatcaggta
     1321 cacagaacca tgcagttcga ggacggggct tctctgaccg taaattatag gtatacttat
     1381 gaaggctcac atattaaggg cgaagcacag gttaaaggaa ccgggtttcc tgcggatggc
     1441 cccgtcatga ctaattctct gacagccgca gattggtgtc gctccaaaaa gacatacccg
     1501 aatgataaga ctataatctc aacattcaaa tggtcctata cgacaggcaa cgggaaacga
     1561 tatagatcca cggctcgaac aacttacaca ttcgctaaac ctatggccgc caattacctc
     1621 aaaaatcagc ccatgtatgt gtttaggaaa accgaattga agcattctaa aacggaactt
     1681 aattttaagg aatggcagaa ggctttcaca gacgtaatgg ggatggatga actctataaa
     1741 tcaggtctcg agtcctcagg gggaacgggt gggtccggag gagttagtaa aggtgaagag
     1801 gacaatatgg caagtttgcc tgcgactcac gagcttcata tctttgggtc tataaatggc
     1861 gttgacttcg atatggttgg ccaaggtact ggcaacccca atgacggtta cgaggagttg
     1921 aatctcaagt ccacaaaagg tgatcttcag ttcagccctt ggattctcgt acctcatatt
     1981 ggatatggct ttcaccagta ccttccatac ccagacggta tgtcaccctt tcaagctgcg
     2041 atggtggatg gttccggcta tcaggtccac cgaacgatgc aattcgagga cggggccagc
     2101 ctcaccgtta attataggta cacctatgag ggaagtcaca taaagggaga agcccaagtg
     2161 aaaggaacag gattcccagc tgatggtcca gtaatgacga actccttgac agcggctgac
     2221 tggtgtagaa gcaaaaagac gtatcctaat gacaagacca tcattagcac tttcaaatgg
     2281 agttatacca caggaaacgg caaacggtac agaagcactg ctagaactac ctacactttc
     2341 gcaaagccga tggctgcaaa ctatttgaag aatcagccca tgtacgtgtt tcgaaaaacg
     2401 gaacttaagc acagtaagac tgaacttaat ttcaaggagt ggcagaaggc gttcacggat
     2461 gtcatgggta tggatgaact gtataaggga gggtctggca ctgggggcac tgccagcagc
     2521 ggatccggtg gcggtgtgag caagggcgag gaggataaca tggcctctct cccagcgaca
     2581 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
     2641 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
     2701 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
     2761 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ataccaagtc
     2821 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
     2881 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     2941 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     3001 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     3061 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     3121 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     3181 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     3241 ggcggaagat ccggtggtgg ttctggtcct cgtccccgtg gtactcgtgg taaaggtcgc
     3301 cgtattcgtc gccgcggtta atctagaggg cccgtttaaa cccgctgatc agcctcgact
     3361 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3421 gaaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3481 gagtaggtgt cattctattc tggggggtgg gggtgggggc aggacagcaa gggggaggat
     3541 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3601 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3661 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3721 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3781 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3841 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3901 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3961 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4021 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4081 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4141 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4201 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4261 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4321 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4381 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4441 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4501 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4561 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4621 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4681 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4741 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4801 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4861 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4921 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4981 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5041 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5101 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5161 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5221 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5281 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5341 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5401 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5461 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5521 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5581 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5641 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5701 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5761 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5821 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6541 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6601 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6661 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6721 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6781 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6841 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6901 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6961 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7021 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7081 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7141 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7201 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7261 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7321 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7381 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7441 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7501 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7561 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7621 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7681 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7741 ccgaaaagtg ccacctgacg tc
//
