LOCUS       sequence-480265-        6159 bp    DNA     circular SYN 24-JUL-2025
DEFINITION  Expresses (n1)QC2-6 FIQ(c4) and FusionRed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6159)
  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 6159)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 24, 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)QC2-6 FIQ(c4)-P2A-FusionRed
FEATURES             Location/Qualifiers
     source          1..6159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     915..937
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     999..1018
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1744
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MASTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTL
                     QYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     CDS             1799..1855
                     /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             1868..2566
                     /codon_start=1
                     /product="monomeric red fluorescent protein optimized for 
                     use as a fusion tag (Shemiakina et al., 2012)"
                     /label=FusionRed
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMAL
                     KLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     misc_feature    2591..3179
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2644..2664)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3062..3073)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3222..3343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3233..3252)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3287..3306
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3722..4310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3802..3821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4481..5272,5273..5341))
                     /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     5104..5123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5342..5446)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5472..5927)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5609..5630)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5821..5840
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5999..6128
                     /label=AAV2 ITR
ORIGIN
        1 atctacgtag ccatgctcta ggaagagtac cattgacgtc aataatgacg tatgttccca
       61 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      121 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      181 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      241 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      301 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      361 aattattttg tgcagcgatg ggggcggggg gggggggggg gggcgcgccg gggggggggg
      421 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggc aggggggggg
      481 ggggggggcg aaaggtttct ttttttgggg gggggggggg ggggggggcg gaggggtgcg
      541 gcggcggcca atcagggcgg cgcgctcgga aagtttcctt ttagggcggg gcggcggcgg
      601 cggcggccct ataaaaggcg aggcgcgcgg cgggcgggag tcgctgcgcg ctgcctccgc
      661 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      721 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      781 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      841 cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc ttcggggggg
      901 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa
      961 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct
     1021 gtctcatcat tttggcaaag aattggatcc ggtaccctaa gccaccatgg ccagcaccgg
     1081 ggaggagctg ttcaccgggg tggtgccctt caagttccag ctgaagggca ccatcaacgg
     1141 caagagcttc accgtggagg gcgagggcga gggcaacagc cacgagggca gccacaaggg
     1201 caagtacgtg tgcaccagcg gcaagctgcc catgagctgg gccgccctgg gcaccagctt
     1261 cggctacggc atgaagtact acaccaagta ccccagcggc ctgaagaact ggttccacga
     1321 ggtgatgccc gagggcttca cctacgacag gcacatccag tacaagggcg acggcagcat
     1381 ccacgccaag caccagcact tcatgaagaa cggcacctac cacaacatcg tggagttcac
     1441 cggccaggac ttcaaggaga acagccccgt gctgaccggc gacatggacg tgagcctgcc
     1501 caacgaggtg cagcacatcc ccatcgacga cggcgtggag tgcaccgtga ccctgcagta
     1561 ccccctgctg agcgacgaga gcaagtgcgt ggaggcctac cagaacacca tcatcaagcc
     1621 cctgcacaac cagcccgccc ccgacgtgcc cttccactgg atcaggaagc agtacaccca
     1681 gagcaaggac gacaccgagg agagggacca catcatccag agcgagaccc tggaggccca
     1741 cctgccatgg cacgagcctt ctgcttctgc cgtgcgaccg gttcacgtgg gaagtggggc
     1801 tacaaacttc tcactgctga aacaggccgg ggatgtggag gaaaacccag gaccacacgt
     1861 gactagtatg gtgagcgagc tgattaagga gaacatgccc atgaagctgt acatggaggg
     1921 caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
     1981 cacccagacc atgagaatca aggtcgtcga gggcggccct ctccccttcg ccttcgacat
     2041 cctggctacc agcttcatgt acggcagcag aaccttcatc aagcaccctc cgggcatccc
     2101 cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga
     2161 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
     2221 caacgtcaag gttagagggg tgaacttccc agccaacggc cctgtgatgc agaagaaaac
     2281 actcggctgg gaggcctcca ccgagacgat gtaccctgct gacggaggac tggaaggcgc
     2341 atgtgacatg gctctgaagc tcgtgggagg aggacacctg atctgcaacc ttgagacaac
     2401 atacagatcc aagaaacccg ctacgaacct caagatgccc ggcgtctaca acgtggacca
     2461 cagactggaa agaatcaagg aggccgacga tgagacatac gtcgagcagc acgaggtggc
     2521 tgtggccaga tactctactg gtggcgctgg tgatggaggt aaataagaat tcgatatcaa
     2581 gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2641 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2701 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2761 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2821 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2881 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2941 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     3001 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3061 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3121 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca
     3181 tcgataccgt cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     3241 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     3301 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     3361 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3421 ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc
     3481 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     3541 tgcgcgctcg ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4141 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5461 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     5521 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     5581 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     5641 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5701 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5761 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5821 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     5881 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     5941 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     6001 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     6061 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     6121 gggttccttg tagttaatga ttaacccgcc atgctactt
//
