LOCUS       sequence-419221-        7343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For labeling the Golgi apparatus. c4 is the appendage for N-terminal
            tagging with td5StayGold. 'GianCreg' is an abbreviation for giantin 
            C-terminal region. '=' denotes a linker [(GGGGS)3]..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7343)
  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 7343)
  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/td5StayGold(c4)=GianCreg
FEATURES             Location/Qualifiers
     source          1..7343
                     /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
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTL
                     LYPLLSDKSKCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETL
                     EAHL"
     promoter        complement(2896..2914)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2897..2914)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2934..2951)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2940..3164
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3210..3638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3297..3316)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3507..3528
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3647..3667)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3652..3981
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3832..3967
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3894..3913
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4048..4842
                     /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(4102..4121)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4712..4731
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5016..5137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5053..5072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5107..5126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5186..5202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5186..5202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5199..5221)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5210..5226
                     /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(5234..5240,5241..5258,5259..5264))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5279..5300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5417..5434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5588..6176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5668..5687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6347..7138,7139..7207))
                     /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     6970..6989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7208..7312)
                     /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 tttaccgggg 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 gacatgaacg tgagcctgcc caacgaggtg cagcacatcc ccagagatga cggcgtggag
     1381 tgcccagtga ccctgctgta ccctctgctg agcgacaaga gcaagtgcgt ggaggcccac
     1441 cagaacacca tctgcaagcc cctgcacaat cagccagccc ccgatgtgcc ataccactgg
     1501 atcagaaagc agtacaccca gagcaaggac gacaccgagg agagagacca catctgccag
     1561 agcgagaccc tggaggccca cctgccatgg cacgagcctt ctgcttctgc cgtggggcat
     1621 ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga caacaacatc
     1681 gatatggtga gtacagggga ggagctgttc accggggtgg tgccatttaa atttcaactt
     1741 aaaggaacca tcaatggcaa atcgtttacc gttgaaggcg aaggtgaagg aaattcacat
     1801 gaaggttctc ataaaggaaa atatgtttgt acaagtggaa aactaccgat gtcatgggca
     1861 gcacttggaa catcctttgg ttatggaatg aaatattata ccaaatatcc tagtggactg
     1921 aagaactggt ttcatgaagt aatgcctgaa ggctttacct acgatcgtca tattcaatat
     1981 aaaggcgatg ggagtatcca tgcaaaacac caacacttta tgaaaaatgg gacttatcac
     2041 aacattgttg aatttactgg tcaggatttt aaagaaaata gtccagtctt aactggagat
     2101 atgaatgtct cattaccgaa tgaagttcaa catataccca gagatgatgg agtagaatgc
     2161 ccagtgacct tgctttatcc tttattatcg gataaatcaa aatgcgttga ggctcaccaa
     2221 aatacaatct gcaagcctct tcataatcaa ccagcacctg atgtcccata tcactggatt
     2281 cgtaaacaat acacacaaag caaagatgat accgaggaac gtgatcatat ttgtcaatca
     2341 gagactctcg aagcacactt accttggcat gagccttcag catcggcagt agaattcgat
     2401 ggtggaggcg gttcaggcgg aggtggctct ggcggtggcg gatcgccatc aagcttatcc
     2461 ggactcagat cacgaggaga accgcagcaa agtttttctg aagctcagca gcagctatgc
     2521 aacaccagac aggaagtgaa tgaattaagg aagctgctgg aagaagaacg agaccaaaga
     2581 gtggctgctg agaatgctct ctctgtggcc gaggagcaga tcagacggtt agagcacagt
     2641 gaatgggact cttcccggac tcctatcatt ggctcctgtg gcactcagga gcaggcactg
     2701 ttaatagatc ttacaagcaa cagttgtcga aggacccgga gtggcgttgg atggaagcga
     2761 gtcctgcgtt cactctgtca ttcacggacc cgagtgccac ttctagcagc catctacttt
     2821 ctaatgattc atgtcctgct cattctgtgt tttacgggcc atctatagct cgagcatgca
     2881 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     2941 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3001 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3061 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3121 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3181 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3241 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3301 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3361 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3421 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3481 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3541 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3601 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3661 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3721 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3781 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3901 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3961 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4021 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4081 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4141 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4201 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4261 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4321 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4381 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4441 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4501 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4561 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4621 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4681 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4741 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4801 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4861 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4921 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4981 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5041 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5101 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5161 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5221 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5281 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5341 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5401 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5461 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5521 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5581 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5641 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5701 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5761 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5821 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5881 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5941 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6001 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6061 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6121 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6181 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6241 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6301 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6361 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6421 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6481 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6541 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6601 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6661 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6721 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6781 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6841 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6901 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6961 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7021 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7081 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7141 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7201 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7261 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7321 cccgaaaagt gccacctgac gtc
//
