LOCUS       sequence-478892-        6087 bp    DNA     circular SYN 17-JUL-2025
DEFINITION  Expresses AausFP1 and FusionRed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6087)
  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 6087)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 17, 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-AausFP1-P2A-FusionRed
FEATURES             Location/Qualifiers
     source          1..6087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     859..881
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     943..962
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1727..1783
                     /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             1796..2494
                     /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    2519..3107
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2572..2592)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2990..3001)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3150..3271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3161..3180)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3215..3234
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3650..4238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3730..3749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4409..5200,5201..5269))
                     /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     5032..5051
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5270..5374)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5400..5855)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5537..5558)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5749..5768
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5927..6056
                     /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 ggggcccccg gggggggggg
      421 gggggggggg gggggggggg gggggggggg cgaggggcgg ggcggggcga ggcggagagg
      481 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      541 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
      601 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
      661 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
      721 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
      781 gggccctttg tgcgggggga gcggctcggg gctgtccgcg gggggacggc tgccttcggg
      841 ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg
      901 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
      961 tgctgtctca tcattttggc aaagaattgg atccggtacc gccaccatga gttacggagc
     1021 acttttgttc agagaaaaaa tcccctatgt tgttgagatg gaaggtgatg ttgagggaat
     1081 gaagttttca gtaagaggta aaggacatgg tgatgctaac actggtaaaa ttgaagccag
     1141 tttcatctgc acaactggtg aactaccagt accttggtca agcattctaa caactgtcac
     1201 atacggtgca cagtgttttg caaaatatcc aaatgatatc aaagactacc caaaaagtgc
     1261 tatgccagaa ggttacgttc aagagcgaac aatcactttt gaaaacgacg gcgtttataa
     1321 gacacgcgct gaagtcacgt acgagaaagg atcagtttat aaccgagtta cgttgaacgg
     1381 ttcaggattt aaaaaaggcg gtaacatctt gggcaaaaaa ttggaattta actacaaccc
     1441 tcattgcatc tatgttcttc cagatgtaca aaataatgga atcaaatgct acatcaacat
     1501 tgttcatgac gtaattggag gaggacaaat aattgctgcg caccaacaat taaatactcc
     1561 tcttggcggt ggaccagtcg acattccaca ctaccatcat atacaagcgc acacaatact
     1621 ttctaaagat cccaaagaaa caagagatca tatgaacgta gtagaggtgt tcagagctat
     1681 cgattgcaaa accgcatatg cacgaccggt tcacgtggga agtggggcta caaacttctc
     1741 actgctgaaa caggccgggg atgtggagga aaacccagga ccacacgtga ctagtatggt
     1801 gagcgagctg attaaggaga acatgcccat gaagctgtac atggagggca ccgtgaacaa
     1861 ccaccacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat
     1921 gagaatcaag gtcgtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag
     1981 cttcatgtac ggcagcagaa ccttcatcaa gcaccctccg ggcatccccg acttctttaa
     2041 gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt
     2101 gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaaggt
     2161 tagaggggtg aacttcccag ccaacggccc tgtgatgcag aagaaaacac tcggctggga
     2221 ggcctccacc gagacgatgt accctgctga cggaggactg gaaggcgcat gtgacatggc
     2281 tctgaagctc gtgggaggag gacacctgat ctgcaacctt gagacaacat acagatccaa
     2341 gaaacccgct acgaacctca agatgcccgg cgtctacaac gtggaccaca gactggaaag
     2401 aatcaaggag gccgacgatg agacatacgt cgagcagcac gaggtggctg tggccagata
     2461 ctctactggt ggcgctggtg atggaggtaa ataagaattc gatatcaagc ttatcgataa
     2521 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2581 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2641 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2701 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2761 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2821 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2881 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     2941 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3001 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3061 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
     3121 acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     3181 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     3241 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     3301 tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     3361 taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa
     3421 tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3481 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4081 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4201 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4261 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4321 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4381 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4441 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4501 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4561 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4621 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4681 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4741 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4801 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4861 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4921 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4981 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5041 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5101 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5161 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5221 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5281 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5341 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     5401 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5461 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5521 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5581 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5641 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5701 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5761 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5821 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     5881 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgc gcgctcgctc
     5941 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
     6001 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta
     6061 gttaatgatt aacccgccat gctactt
//
