LOCUS       sequence-480639-        6847 bp    DNA     circular SYN 28-JUL-2025
DEFINITION  Expresses (n1)StayGold(c4) and mCherry in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6847)
  AUTHORS   Zhang H, Lesnov GD, Subach OM, Zhang W, Kuzmicheva TP, Vlaskina AV, 
            Samygina VR, Chen L, Ye X, Nikolaeva AY, Gabdulkhakov A, Papadaki S,
            Qin W, Borshchevskiy V, Perfilov MM, Gavrikov AS, Drobizhev M, 
            Mishin AS, Piatkevich KD, Subach FV
  TITLE     Bright and stable monomeric green fluorescent protein derived from 
            StayGold.
  JOURNAL   Nat Methods. 2024 Apr;21(4):657-665. doi: 
            10.1038/s41592-024-02203-y. Epub 2024 Feb 26.
  PUBMED    38409224
REFERENCE   2  (bases 1 to 6847)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-024-02203-y"; journalName: "Nat Methods"; date: 
            "2024-04"; volume: "21"; issue: "4"; pages: "657-665"
COMMENT     Sequence Label: pAAV-CAG-(n1)StayGold(c4)-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..6847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    2..123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13..32)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     67..86
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(502..1090)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(582..601)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1261..2052,2053..2121))
                     /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     1884..1903
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2122..2226)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2252..2707)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2389..2410)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2601..2620
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2779..2908
                     /label=AAV2 ITR
     gap             3385..3394
                     /estimated_length=10
     repeat_region   3529..3658
                     /label=AAV2 ITR
     primer_bind     4531..4553
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4615..4634
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      4673..4682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5411..5467
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5480..6190
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5625..5643)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5893..5912
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6218..6806
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6271..6291)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6689..6700)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
       61 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
      121 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
      181 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
      241 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
      301 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gactcgctgc gctcggtcgt
      361 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      421 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      481 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      541 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      601 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      661 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      721 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      781 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      841 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      901 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
      961 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1021 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1081 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1141 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1201 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1261 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1321 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1381 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     1441 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1501 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1561 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1621 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1681 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1741 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1801 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1861 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1921 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1981 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2041 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2101 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2161 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2221 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     2281 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2341 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2401 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2461 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2521 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2581 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     2641 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     2701 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     2761 tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
     2821 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     2881 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
     2941 tctacgtagc catgctctag gaagagtacc attgacgtca ataatgacgt atgttcccat
     3001 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     3061 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     3121 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     3181 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
     3241 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
     3301 attattttgt gcagcgatgg gggcgggggg gggggggggg ggcgcgccgg gggggggggg
     3361 gggggggggg gggggggggg ggggnnnnnn nnnntagcgg tcccgctgcg cgtacccccc
     3421 ccccccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     3481 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc
     3541 tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc
     3601 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg
     3661 tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag gaagagtacc
     3721 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     3781 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     3841 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     3901 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     3961 taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     4021 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     4081 gggggggggg ggcgcgccgg gggggggggg gggggggggg gggggggggc ggggcggggc
     4141 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
     4201 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
     4261 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
     4321 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     4381 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
     4441 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg
     4501 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     4561 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     4621 tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatccggta ccgccaccat
     4681 ggctagtaca ggggaggagc tgttcaccgg ggtggtgcca tttaaatttc aacttaaagg
     4741 aaccatcaat ggcaaatcgt ttaccgttga aggcgaaggt gaaggaaatt cacatgaagg
     4801 ttctcataaa ggaaaatatg tttgtacaag tggaaaacta ccgatgtcat gggcagcact
     4861 tggaacatcc tttggttatg gaatgaaata ttataccaaa tatcctagtg gactgaagaa
     4921 ctggtttcat gaagtaatgc ctgaaggctt tacctacgat cgtcatattc aatataaagg
     4981 cgatgggagt atccatgcaa aacaccaaca ctttatgaaa aatgggactt atcacaacat
     5041 tgttgaattt actggtcagg attttaaaga aaatagtcca gtcttaactg gagatatgaa
     5101 tgtctcatta ccgaatgaag ttcaacatat acccagagat gatggagtag aatgcccagt
     5161 gaccttgctt tatcctttat tatcggataa atcaaaatgc gttgaggctc accaaaatac
     5221 aatctgcaag cctcttcata atcaaccagc acctgatgtc ccatatcact ggattcgtaa
     5281 acaatacaca caaagcaaag atgataccga ggaacgtgat catatttgtc aatcagagac
     5341 tctcgaagca cacttaccat ggcacgagcc ttctgcttct gccgtgcgac cggttcacgt
     5401 gggaagtggg gctacaaact tctcactgct gaaacaggcc ggggatgtgg aggaaaaccc
     5461 aggaccacac gtgactagta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
     5521 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     5581 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     5641 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     5701 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     5761 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     5821 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     5881 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     5941 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     6001 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     6061 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     6121 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     6181 gtacaagtag taagaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     6241 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6301 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6361 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     6421 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     6481 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6541 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6601 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6661 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6721 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6781 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc
     6841 agacatg
//
