LOCUS       sequence-463670-        3581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mGold2s in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3581)
  AUTHORS   Lee J, Lai S, Yang S, Zhao S, Blanco FA, Lyons AC, Merino-Urteaga R,
            Ahrens JF, Nguyen NA, Liu H, Liu Z, Lambert GG, Shaner NC, Chen L, 
            Tolias KF, Zhang J, Ha T, St-Pierre F
  TITLE     Bright and photostable yellow fluorescent proteins for extended 
            imaging.
  JOURNAL   Nat Commun. 2025 Apr 4;16(1):3241. doi: 10.1038/s41467-025-58223-5.
  PUBMED    40185748
REFERENCE   2  (bases 1 to 3581)
  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 
            Commun."; date: "2025-04-4"; volume: "16(1):3241. doi"; pages: " 
            10.1038/s41467-025-58223-5"
COMMENT     Sequence Label: pNCS-mGold2s
FEATURES             Location/Qualifiers
     source          1..3581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(199..201,202..204,205..918)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFEDD
                     GNYKTRAEVKFEGGTLVNRVELKGIDFKEDGNILGHKLEYNYNCHNVFITADRQQNGIK
                     VNFKIRHNIEVGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLSMDELYK"
     primer_bind     complement(505..524)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     852..873
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(982..1000)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      996..1043
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1140..1595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1227..1246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1437..1458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1621..1725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1726..1794,1795..2586)
                     /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     complement(1944..1963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2757..3345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3246..3265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3499..3516
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatccgat ggcgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      241 atcctggtcg agctggacgg cgacataaac ggccacaagt tcagcgtgtc cggcgagggc
      301 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      361 cccgtgccct ggcccaccct cgtgaccagc ctgggctacg gcctgcagtg cttcgcccgc
      421 taccccgaca acatgaagcg gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      481 caggagcgca ccatcttctt cgaggacgac ggcaactaca agacccgcgc cgaggtgaag
      541 tttgagggcg gcaccctggt gaaccgcgtc gagctgaagg gcatcgactt caaggaggac
      601 ggcaacatcc tggggcacaa gctggagtac aactacaact gccacaacgt ctttatcacc
      661 gccgacaggc agcagaacgg catcaaggtc aacttcaaga tccgccacaa catcgaggtc
      721 ggcggcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      781 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
      841 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcagcatg
      901 gacgagctgt acaagtaatg aattcgaagc ttgatccggc tgctaacaaa gcccgaaagg
      961 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     1021 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatct ggcgtaatag
     1081 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     1141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1501 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1561 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     1621 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     1681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     1741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     1861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     1921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     1981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2281 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     2341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     2641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     2701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     2761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2881 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3421 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3481 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3541 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca g
//
