LOCUS       sequence-417677-        5951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the bright and photostable StayGold 
            fluorescent protein. Codon optimised for expression in Escherichia 
            coli. Contains a N-terminal 10xHis tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5951)
  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 5951)
  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: pET-MCN-10His-StayGold
FEATURES             Location/Qualifiers
     source          1..5951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(473..1555)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1517..1536
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1556..1633)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1839..1858
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1942..1961
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1942..1960
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1961..1985
                     /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)."
     RBS             2000..2022
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2039..2068
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2075..2725
                     /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"
     primer_bind     complement(2827..2845)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2841..2888
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2925..3380
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3012..3031)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3222..3243
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3472..4287)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3472..4287)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4195..4214
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4279..4298
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4409..4997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4898..4917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5151..5168
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5183..5322
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5248..5267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5408..5430
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5424..5615)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
       61 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
      121 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
      181 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
      241 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
      301 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
      361 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
      421 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
      481 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      541 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
      601 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
      661 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
      721 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
      781 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
      841 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
      901 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
      961 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     1021 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     1081 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     1141 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     1201 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     1261 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     1321 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     1381 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     1441 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     1501 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     1561 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     1621 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     1681 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     1741 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     1801 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     1861 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     1921 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     1981 attccccact agtaataatt ttgtttaact ttaagaagga gatataatgg gcagcagcca
     2041 tcaccatcac catcaccatc accatcacgg ttctatggcc agcacgccgt ttaagttcca
     2101 gttaaagggg acaattaacg ggaaaagctt tacagtcgag ggtgaagggg aagggaactc
     2161 gcatgaaggt agtcataaag gtaagtatgt gtgtacgagc ggaaagttgc cgatgtcttg
     2221 ggcggctctt ggtacttcgt ttgggtacgg aatgaaatac tatacaaaat acccgtcggg
     2281 cctgaaaaat tggttccatg aggtgatgcc cgaggggttc acgtatgatc gccacatcca
     2341 gtacaaaggt gatggctcaa tccacgcaaa gcaccaacac ttcatgaaga atgggactta
     2401 tcataacatt gtcgagttta ctggccagga cttcaaggaa aacagccccg tattgacagg
     2461 tgacatgaat gtgagcttgc cgaacgaggt gcagcatatt ccacgtgacg acggagttga
     2521 atgtcccgtc acattgttat accccttgct tagcgataaa tcaaagtgtg tcgaagcaca
     2581 tcaaaacact atctgcaaac ctcttcataa ccaacctgcg ccagatgtac cgtaccattg
     2641 gattcgcaaa caatatacac agtctaagga tgatacggag gaacgcgatc atatctgtca
     2701 gtccgagacg ttagaagctc atctttaact cgagcttaag caattgggat cctaatagtc
     2761 tagagctagc cctaggagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     2821 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2881 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     2941 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3001 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3061 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3121 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3181 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3241 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3301 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3361 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
     3481 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     3541 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     3601 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     3661 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     3721 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     3781 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     3841 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     3901 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     3961 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     4021 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     4081 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     4141 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     4201 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     4261 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     4321 cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4381 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4441 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4501 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4561 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4621 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4681 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4741 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4801 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4861 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4921 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4981 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5041 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5101 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5161 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5221 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5281 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     5341 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5401 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     5461 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     5521 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     5581 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     5641 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     5701 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     5761 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     5821 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     5881 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     5941 gttgctcagg t
//
