LOCUS       sequence-309932-        6816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mGold in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  AUTHORS   Lee J, Liu Z, Suzuki PH, Ahrens JF, Lai S, Lu X, Guan S, St-Pierre F
  TITLE     Versatile phenotype-activated cell sorting.
  JOURNAL   Sci Adv. 2020 Oct 23;6(43). pii: 6/43/eabb7438. doi: 
            10.1126/sciadv.abb7438. Print 2020 Oct.
  PUBMED    33097540
REFERENCE   2  (bases 1 to 6816)
  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: "Sci Adv."; 
            date: "2020-10-23"; volume: "6(43). pii: 6/43/eabb7438. doi"; pages:
            " 10.1126/sciadv.abb7438. Print 2020 Oct"
COMMENT     Sequence Label: pCaggs-mGold
FEATURES             Location/Qualifiers
     source          1..6816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..37
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     99..118
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      251..260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(257..259,260..262,263..976)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(302..323)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(563..582)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     910..931
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1091..1108)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1097..1321
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1367..1795
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1454..1473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1664..1685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1804..1824)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1809..2138
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1989..2124
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2051..2070
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2149..2748
                     /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(2149..2168)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2645..2665
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2908..3029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2945..2964)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2999..3018
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3078..3094)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3078..3094)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3091..3113)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3102..3118
                     /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(3126..3132,3133..3150,3151..3156))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3171..3192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3309..3326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3480..4068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3560..3579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4239..5030,5031..5099))
                     /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     4862..4881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5100..5204)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5236..5615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          join(5891..6816,1..91)
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
ORIGIN
        1 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa
       61 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct
      121 gtctcatcat tttggcaaag aattcctcga ggaattcact cctcaggtgc aggctgccta
      181 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctgca
      241 tgagctagcc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      301 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      361 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      421 cgtgccctgg cccaccctcg tgaccagcct gggctacggc ctgcagtgct tcgcccgcta
      481 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      541 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      601 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      661 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc
      721 cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg
      781 cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      841 gctgcccgac aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa
      901 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      961 cgagctgtac aagtaatgaa agcttggtac cgagctcgga tccactagtc cagtgtggtg
     1021 gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc
     1081 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1141 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1201 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1261 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1321 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     1381 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1441 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1501 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1561 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1621 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1681 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1741 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     1801 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     1861 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     1921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     1981 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2041 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2101 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccgcat gaccgagtac
     2161 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     2221 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     2281 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     2341 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     2401 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     2461 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     2521 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     2581 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     2641 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     2701 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatt cgaaatgacc
     2761 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     2821 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     2881 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     2941 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3001 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     3061 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3121 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3181 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3241 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3301 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3361 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4141 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4201 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4261 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4321 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4381 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4441 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4501 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4561 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4621 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4681 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4741 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4801 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4861 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4921 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4981 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5041 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5101 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5161 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5221 gtgccacctg acgtcgacat tgattattga ctagttatta atagtaatca attacggggt
     5281 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     5341 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     5401 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     5461 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     5521 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     5581 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
     5641 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
     5701 tattttgtgc agcgatgggg gcgggggggg ggggggggcg ccrggsgggg gggggggggg
     5761 gggggggsgg ggsggggsgr ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     5821 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     5881 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     5941 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     6001 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
     6061 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
     6121 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
     6181 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag
     6241 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     6301 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     6361 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     6421 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc
     6481 gggcggggcg gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg
     6541 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     6601 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg
     6661 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat
     6721 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg
     6781 gggctgtccg cggggggacg gctgccttcg gggggg
//
