LOCUS       sequence-419202-        6984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For filamentous actin labeling. Alternatively, the UtrCH gene can be
            replaced with a target gene for N-terminal tagging with mStayGold 
            via a c4 adaptor and a linker [(GGGGS)3] (denoted as '=')..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6984)
  AUTHORS   Ando R, Shimozono S, Ago H, Takagi M, Sugiyama M, Kurokawa H, Hirano
            M, Niino Y, Ueno G, Ishidate F, Fujiwara T, Okada Y, Yamamoto M, 
            Miyawaki A
  TITLE     StayGold variants for molecular fusion and membrane-targeting 
            applications.
  JOURNAL   Nat Methods. 2023 Nov 30. doi: 10.1038/s41592-023-02085-6.
  PUBMED    38036853
REFERENCE   2  (bases 1 to 6984)
  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: "Nat 
            Methods. 2023 Nov 30. doi: 10.1038/s41592-023-02085-6."
COMMENT     Sequence Label: pcDNA3/mStayGold(c4)=UtrCH
FEATURES             Location/Qualifiers
     source          1..6984
                     /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"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             907..1584
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTL
                     QYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     primer_bind     1616..1632
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1617..1633
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2537..2555)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2538..2555)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2575..2592)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2581..2805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2851..3279
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2938..2957)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3148..3169
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3288..3308)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3293..3622
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3473..3608
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3535..3554
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3689..4483
                     /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(3743..3762)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4353..4372
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4657..4778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4694..4713)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4748..4767
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4827..4843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4827..4843)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4840..4862)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4851..4867
                     /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(4875..4881,4882..4899,4900..4905))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4920..4941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5058..5075)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5229..5817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5309..5328)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5988..6779,6780..6848))
                     /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     6611..6630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6849..6953)
                     /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 cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gccaccatgg tgtctacagg cgaggagctg tttaccggcg tggtgccctt caagttccag
      961 ctgaagggca ccatcaacgg caagagcttc accgtggaag gcgagggcga gggcaatagc
     1021 cacgagggca gccacaaagg caagtacgtg tgcaccagcg gcaaactgcc aatgtcttgg
     1081 gccgccctgg gaactagctt cggctatggc atgaagtact acaccaagta ccccagcggc
     1141 ctgaagaact ggttccacga ggtgatgccc gagggcttca cctacgacag acacatccag
     1201 tacaagggcg acggcagcat ccacgccaag caccagcact tcatgaagaa cggcacctac
     1261 cacaacatcg tggagttcac cggccaggac ttcaaggaga acagccccgt gctgaccggc
     1321 gacatggacg tgagcctgcc caacgaggtg cagcacatcc ccatagatga cggcgtggag
     1381 tgcacagtga ccctgcagta ccctctgctg agcgacgaga gcaagtgcgt ggaggcctac
     1441 cagaacacca tcatcaagcc cctgcacaat cagccagccc ccgatgtgcc attccactgg
     1501 atcagaaagc agtacaccca gagcaaggac gacaccgagg agagagacca catcatccag
     1561 agcgagaccc tggaggccca cctgccatgg cacgagcctt ctgcttctgc cgtgctcgag
     1621 gtcgacggta tcgataagct tgatggtgga ggcggttcag gcggaggtgg ctctggcggt
     1681 ggcggatcga tcgaattcct gcagcccggg ggatccatgg ccaagtatgg agaacatgaa
     1741 gccagtcctg acaatgggca gaacgaattc agtgatatca ttaagtccag atctgatgaa
     1801 cacaatgacg tacagaagaa aacctttacc aaatggataa atgctcgatt ttcaaagagt
     1861 gggaaaccac ccatcaatga tatgttcaca gacctcaaag atggaaggaa gctattggat
     1921 cttctagaag gcctcacagg aacatcactg ccaaaggaac gtggttccac aagggtacat
     1981 gccttaaata acgtcaacag agtgctgcag gttttacatc agaacaatgt ggaattagtg
     2041 aatatagggg gaactgacat tgtggatgga aatcacaaac tgactttggg gttactttgg
     2101 agcatcattt tgcactggca ggtgaaagat gtcatgaagg atgtcatgtc ggacctgcag
     2161 cagacgaaca gtgagaagat cctgctcagc tgggtgcgtc agaccaccag gccctacagc
     2221 caagtcaacg tcctcaactt caccaccagc tggacagatg gactcgcctt taatgctgtc
     2281 ctccaccgac ataaacctga tctcttcagc tgggataaag ttgtcaaaat gtcaccaatt
     2341 gagagacttg aacatgcctt cagcaaggct caaacttatt tgggaattga aaagctgtta
     2401 gatcctgaag atgttgccgt tcagcttcct gacaagaaat ccataattat gtatttaaca
     2461 tctttgtttg aggtgctacc tcagcaagtc accatagact aagcggccgc tcgagcatgc
     2521 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
     2581 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2641 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2701 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2761 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     2821 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     2881 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2941 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3001 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3061 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3121 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3181 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3241 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     3301 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3361 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3421 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3481 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3541 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3601 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3661 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3721 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3781 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3841 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3901 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3961 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4021 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4081 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4141 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4201 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4261 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4321 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4381 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4441 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4501 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4561 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4621 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4681 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4741 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4801 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4861 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4921 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4981 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5041 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5101 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5161 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5221 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5281 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5341 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5401 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5461 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5521 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5581 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5641 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5701 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5761 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5821 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5881 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5941 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6001 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6061 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6121 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6181 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6241 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6301 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6361 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6421 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6481 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6541 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6601 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6661 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6721 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6781 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6841 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6901 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6961 ccccgaaaag tgccacctga cgtc
//
