pAAV-CAG-Electra2-P2A-GFP

LOCUS       pAAV-CAG-Electra2-P2A-GFP           6244 bp DNA     circular SYN 29-MAY-2023
DEFINITION  Expresses TagRFP658 and GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6244)
  TITLE     Rapid Directed Molecular Evolution of Fluorescent Proteins in 
            Mammalian Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6244)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 29, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
FEATURES             Location/Qualifiers
     source          1..6244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        279..556
                     /label=chicken beta-actin promoter
     CDS             983..1693
                     /codon_start=1
                     /label=Electra2
                     /translation="MVSKGEELIEENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFLFGSKTFIKYPADIPDFFEQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGGLVYNVKVRGVNFHSKGPVMQKKTEGWEPFTEMMYPADGGLRGYTD
                     IALKVDGGGHLHANIVTTYRSKKTVGNIKMPGVHAVDYRLERIEESDNETYVVLREVAV
                     AKYSNLGGGMDELFK"
     regulatory      983..986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1718..1774
                     /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             1787..2503
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGRLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    2528..3116
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2999..3010)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3159..3280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3442..3571
                     /label=AAV2 ITR
     rep_origin      complement(3808..4396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4567..5427)
                     /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"
     promoter        complement(5428..5532)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5558..6013)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   6085..6214
                     /label=AAV2 ITR
ORIGIN
        1 tctacgtagc catgctctag gaagagtacc attgacgtca ataatgacgt atgttcccat
       61 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      121 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      181 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      241 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
      301 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
      361 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg
      421 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
      481 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
      541 aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc
      601 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      661 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      721 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      781 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
      841 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
      901 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
      961 aattggatcc ggtaccgcca ccatggtgtc taagggcgaa gagctgatcg aggaaaatat
     1021 gcgtatgaag gtggtcatgg aaggttcggt caacggccac caattcaaat gcacaggtga
     1081 aggagaaggc agaccgtacg agggagtgca aaccatgagg atcaaagtca tcgagggagg
     1141 acccctgcca tttgcctttg acattcttgc cacgtcgttc ctgttcggca gcaagacctt
     1201 tatcaagtac ccggccgaca tccctgattt cttcgaacag tcctttcctg agggctttac
     1261 ttgggaaaga gttacgagat acgaagatgg tggagtagtc accgtcacgc aggacaccag
     1321 ccttgaggat ggtgggctcg tctacaacgt caaggtcaga ggggtaaact ttcactccaa
     1381 gggtcccgtg atgcagaaga agaccgaggg ttgggagcct ttcacagaga tgatgtatcc
     1441 agcagatggt ggtctgagag gatataccga catcgcactg aaagttgatg gtggtggcca
     1501 tctgcacgcc aacatcgtga caacttacag gtcaaaaaag accgtcggga acatcaagat
     1561 gcccggtgtc catgccgttg attaccgcct ggaaaggatc gaagagagtg acaatgaaac
     1621 ctacgtagtg ctaagagaag tggcagttgc caaatacagc aaccttggtg gtggcatgga
     1681 cgagctgttc aagcgaccgg ttcacgtggg aagtggggct acaaacttct cactgctgaa
     1741 acaggccggg gatgtggagg aaaacccagg accacacgtg actagtgtga gcaagggcga
     1801 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1861 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1921 gttcatctgc accaccggca ggctgcccgt gccctggccc accctcgtga ccaccctgac
     1981 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2041 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2101 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2161 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2221 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2281 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2341 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     2401 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2461 cgccgccggg atcactctcg gcatggacga gctgtacaag taagaattcg atatcaagct
     2521 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2581 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2641 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2701 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2761 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2821 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2881 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2941 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3001 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3061 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3121 ataccgtcga cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3181 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3241 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3301 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta
     3361 caaatgtggt aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg
     3421 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     3481 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3541 gggcggcctc agtgagcgag cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg
     3601 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3661 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3721 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3781 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3841 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3901 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3961 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4021 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4081 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4141 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4201 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4261 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4321 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4381 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4441 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4501 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4561 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4621 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4681 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4741 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4801 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4861 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4921 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4981 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5041 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5101 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5161 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5221 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5281 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5341 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5401 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5461 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5521 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     5581 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     5641 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     5701 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     5761 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     5821 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     5881 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     5941 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     6001 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6061 cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc cgcccgggca
     6121 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     6181 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc
     6241 tact
//