LOCUS       sequence-417675-        6071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the bright and photostable StayGold 
            fluorescent protein. Contains a N-terminal 10xHis tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  AUTHORS   Ivorra-Molla E, Akhuli D, McAndrew MBL, Scott W, Kumar L, Palani S, 
            Mishima M, Crow A, Balasubramanian MK
  TITLE     A monomeric StayGold fluorescent protein.
  JOURNAL   Nat Biotechnol. 2023 Dec 11. doi: 10.1038/s41587-023-02018-w.
  PUBMED    38081970
REFERENCE   2  (bases 1 to 6071)
  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 
            Biotechnol. 2023 Dec 11. doi: 10.1038/s41587-023-02018-w."
COMMENT     Sequence Label: pcDNA3-10His-StayGold
FEATURES             Location/Qualifiers
     source          1..6071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(104..895,896..964))
                     /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     727..746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(965..1069)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1144..1163)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1335..1714
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1715..1918
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1869..1889
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1963..1982
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1963..1981
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1995..2004
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2013..2042
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2049..2699
                     /codon_start=1
                     /product="bright and highly photostable dimeric green 
                     fluorescent protein (Hirano et al., 2022)"
                     /label=StayGold
                     /note="engineered variant of a GFP from Cytaeis uchidae"
                     /translation="MASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKL
                     PMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFM
                     KNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTLLYPLLSDKS
                     KCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETLEAHL"
     promoter        complement(2724..2742)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2725..2742)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2762..2779)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2768..2992
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3038..3466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3125..3144)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3335..3356
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3475..3495)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3480..3809
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3660..3795
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3722..3741
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3876..4670
                     /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(3930..3949)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4540..4559
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4844..4965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4881..4900)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4935..4954
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5014..5030)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5014..5030)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5027..5049)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5038..5054
                     /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(5062..5068,5069..5086,5087..5092))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5107..5128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5245..5262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5416..6004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5496..5515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
       61 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      121 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      181 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      241 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
      301 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      361 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      421 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      481 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
      541 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
      601 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
      661 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
      721 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
      781 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
      841 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
      901 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
      961 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1021 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1081 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     1141 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     1201 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     1261 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     1321 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     1381 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1441 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1501 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1561 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1621 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1681 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1741 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     1801 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     1861 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     1921 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
     1981 ggagacccaa gcttgccacc atgggcagca gccatcacca tcaccatcac catcaccatc
     2041 acggttctat ggccagcacg ccgtttaagt tccagttaaa ggggacaatt aacgggaaaa
     2101 gctttacagt cgagggtgaa ggggaaggga actcgcatga aggtagtcat aaaggtaagt
     2161 atgtgtgtac gagcggaaag ttgccgatgt cttgggcggc tcttggtact tcgtttgggt
     2221 acggaatgaa atactataca aaatacccgt cgggcctgaa aaattggttc catgaggtga
     2281 tgcccgaggg gttcacgtat gatcgccaca tccagtacaa aggtgatggc tcaatccacg
     2341 caaagcacca acacttcatg aagaatggga cttatcataa cattgtcgag tttactggcc
     2401 aggacttcaa ggaaaacagc cccgtattga caggtgacat gaatgtgagc ttgccgaacg
     2461 aggtgcagca tattccacgt gacgacggag ttgaatgtcc cgtcacattg ttatacccct
     2521 tgcttagcga taaatcaaag tgtgtcgaag cacatcaaaa cactatctgc aaacctcttc
     2581 ataaccaacc tgcgccagat gtaccgtacc attggattcg caaacaatat acacagtcta
     2641 aggatgatac ggaggaacgc gatcatatct gtcagtccga gacgttagaa gctcatcttt
     2701 aacatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     2761 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2821 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2881 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2941 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3001 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3061 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3121 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3181 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3241 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3301 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3361 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3421 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3481 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3541 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3601 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3661 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3721 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3781 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3841 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3901 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3961 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4021 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     4081 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4141 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4201 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4261 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4321 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4381 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4441 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4501 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4561 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4621 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4681 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4741 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4801 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4861 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4921 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4981 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5041 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5101 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5161 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5221 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5281 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5341 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5821 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6061 ttaagggatt t
//
