LOCUS       sequence-470111-        6819 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mGold2t in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6819)
  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 6819)
  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: pCaggs-mGold2t
FEATURES             Location/Qualifiers
     source          1..6819
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      8..17
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(14..16,17..19,20..733)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFEDD
                     GNYKTRAEVKFEGGTLVNRVELKGIDFKEDGNILGHKLEYNYNCHNVFITADRQQNGIK
                     VNFKIRHNIEVGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQTKLSKDPNEKRDHMVLL
                     EFVTAAGITLSMDELYK"
     primer_bind     complement(320..339)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     667..688
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(848..865)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    854..1078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1124..1552
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1211..1230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1421..1442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1561..1581)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1566..1895
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1746..1881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1808..1827
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1906..2505
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1906..1925)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2402..2422
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2665..2786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2702..2721)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2756..2775
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2835..2851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2835..2851)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2848..2870)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2859..2875
                     /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(2883..2889,2890..2907,2908..2913))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2928..2949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3066..3083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3237..3825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3317..3336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3996..4787,4788..4856))
                     /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     4619..4638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4857..4961)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4993..5372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5374..5650
                     /label=chicken beta-actin promoter
     intron          5651..6667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     6591..6613
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     6675..6694
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gctagccgcc accatggcga gcaagggcga ggagctgttc accggggtgg tgcccatcct
       61 ggtcgagctg gacggcgaca taaacggcca caagttcagc gtgtccggcg agggcgaggg
      121 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      181 gccctggccc accctcgtga ccagcctggg ctacggcctg cagtgcttcg cccgctaccc
      241 cgacaacatg aagcggcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      301 gcgcaccatc ttcttcgagg acgacggcaa ctacaagacc cgcgccgagg tgaagtttga
      361 gggcggcacc ctggtgaacc gcgtcgagct gaagggcatc gacttcaagg aggacggcaa
      421 catcctgggg cacaagctgg agtacaacta caactgccac aacgtcttta tcaccgccga
      481 caggcagcag aacggcatca aggtcaactt caagatccgc cacaacatcg aggtcggcgg
      541 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      601 gcccgacaac cactacctga gctaccagac caagctgagc aaagacccca acgagaagcg
      661 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctca gcatggacga
      721 gctgtacaag taatgaaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      781 ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc
      841 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
      901 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
      961 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1021 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     1081 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     1141 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1201 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1261 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1321 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1381 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1441 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1501 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     1561 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     1621 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     1681 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     1741 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     1801 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     1861 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgcatgac cgagtacaag
     1921 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     1981 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     2041 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     2101 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     2161 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     2221 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     2281 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     2341 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     2401 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     2461 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgattcga aatgaccgac
     2521 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     2581 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     2641 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     2701 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2761 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     2821 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2881 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2941 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3001 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3061 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     3841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     3901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     3961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     4861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     4921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     4981 ccacctgacg tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     5041 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     5101 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     5161 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     5221 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     5281 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     5341 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     5401 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     5461 tttgtgcagc gatgggggcg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     5521 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     5581 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     5641 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     5701 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     5761 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
     5821 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
     5881 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
     5941 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag
     6001 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     6061 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     6121 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     6181 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc
     6241 gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg
     6301 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     6361 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg
     6421 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat
     6481 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg
     6541 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
     6601 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     6661 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     6721 cctcgaggaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     6781 caatgccctg gctcacaaat accactgaga tctgcatga
//
