LOCUS       sequence-464049-        3581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mGold2t 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-mGold2t
FEATURES             Location/Qualifiers
     source          1..3581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      514..1102
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1003..1022
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1256..1273
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1358..1377
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1358..1376
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1408..1430
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1450..1467
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1471..1503
                     /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(1507..1515,1516..1530)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(1537..1539,1540..1542,1543..2256)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRVVLKGIDFKEDGNIMGHKLEYNYNCHNVFITADRQQNGIK
                     VNFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYLTKLSKDPNEKRDHMVLL
                     EFVTAAGITLSMDELYK"
     primer_bind     complement(1843..1862)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2190..2211
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2320..2338)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2334..2381
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2478..2933
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2565..2584)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2775..2796
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2959..3063
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3064..3132,3133..3581,1..343)
                     /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(3282..3301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
       61 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      121 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      181 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
      241 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
      301 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
      361 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
      421 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
      481 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
      541 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      601 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      661 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      721 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      781 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      841 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
      901 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
      961 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1021 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1081 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1141 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     1201 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     1261 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1321 gccgattcat taatgcagga tctcgatccc gcgaaattaa tacgactcac tatagggaga
     1381 ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     1441 cggggttctc atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg
     1501 ggtcgggatc tgtacgacga tgacgataag gatccgatgg cgagcaaggg cgaggagctg
     1561 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acataaacgg ccacaagttc
     1621 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     1681 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccagcct gggctacggc
     1741 ctgcagtgct tcgcccgcta ccccgacaac atgaagcggc acgacttctt caagtccgcc
     1801 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     1861 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcgtcgt gctgaagggc
     1921 atcgacttca aggaggacgg caacatcatg gggcacaagc tggagtacaa ctacaactgc
     1981 cacaacgtct ttatcaccgc cgacaggcag cagaacggca tcaaggtcaa cttcaagatc
     2041 cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     2101 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacct gaccaagctg
     2161 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2221 gggatcactc tcagcatgga cgagctgtac aagtaatgaa ttcgaagctt gatccggctg
     2281 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     2341 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     2401 ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2461 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2521 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2581 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2641 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2701 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2761 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2821 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2881 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     2941 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3001 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3061 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3121 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3181 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3241 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3301 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3361 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3421 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3481 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3541 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa a
//
