pAAV-CAG-mStayGold(E138D)-P2A-FusionRed

LOCUS       pAAV-CAG-mStayGold(E138D)-P2A-FusionRed             6166 bp DNA     circular SYN 01-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6166)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Jul 1, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        279..556
                     /label=chicken beta-actin promoter
     misc_feature    977..982
                     /label=Kozak sequence
     CDS             983..1633
                     /codon_start=1
                     /label=mStayGold(E138D)
                     /translation="MASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKL
                     PMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFM
                     KNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNDVQHIPRDDGVECPVTLLYPLLSDKS
                     KCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETLEAHL"
     primer_bind     1375..1430
                     /label=0707-2-E138D-F
     misc_feature    1394..1396
                     /label=E138D
     CDS             1658..1714
                     /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             1727..2422
                     /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"
     primer_bind     complement(2402..2446)
                     /label=0714-FusionRed-R
     misc_feature    2450..3038
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2921..2932)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3081..3202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   3364..3493
                     /label=AAV2 ITR
     rep_origin      complement(3730..4318)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4489..5349)
                     /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(5350..5454)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5480..5935)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     repeat_region   6007..6136
                     /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 ccatggctag tacaccattt aaatttcaac ttaaaggaac
     1021 catcaatggc aaatcgttta ccgttgaagg cgaaggtgaa ggaaattcac atgaaggttc
     1081 tcataaagga aaatatgttt gtacaagtgg aaaactaccg atgtcatggg cagcacttgg
     1141 aacatccttt ggttatggaa tgaaatatta taccaaatat cctagtggac tgaagaactg
     1201 gtttcatgaa gtaatgcctg aaggctttac ctacgatcgt catattcaat ataaaggcga
     1261 tgggagtatc catgcaaaac accaacactt tatgaaaaat gggacttatc acaacattgt
     1321 tgaatttact ggtcaggatt ttaaagaaaa tagtccagtc ttaactggag atatgaatgt
     1381 ctcattaccg aatgacgttc aacatatacc cagagatgat ggagtagaat gcccagtgac
     1441 cttgctttat cctttattat cggataaatc aaaatgcgtt gaggctcacc aaaatacaat
     1501 ctgcaagcct cttcataatc aaccagcacc tgatgtccca tatcactgga ttcgtaaaca
     1561 atacacacaa agcaaagatg ataccgagga acgtgatcat atttgtcaat cagagactct
     1621 cgaagcacac ttacgaccgg ttcacgtggg aagtggggct acaaacttct cactgctgaa
     1681 acaggccggg gatgtggagg aaaacccagg accacacgtg actagtatgg tgagcgagct
     1741 gattaaggag aacatgccca tgaagctgta catggagggc accgtgaaca accaccactt
     1801 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa
     1861 ggtcgtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta
     1921 cggcagcaga accttcatca agcaccctcc gggcatcccc gacttcttta agcagtcctt
     1981 ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc
     2041 tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaagg ttagaggggt
     2101 gaacttccca gccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggcctccac
     2161 cgagacgatg taccctgctg acggaggact ggaaggcgca tgtgacatgg ctctgaagct
     2221 cgtgggagga ggacacctga tctgcaacct tgagacaaca tacagatcca agaaacccgc
     2281 tacgaacctc aagatgcccg gcgtctacaa cgtggaccac agactggaaa gaatcaagga
     2341 ggccgacgat gagacatacg tcgagcagca cgaggtggct gtggccagat actctactgg
     2401 tggcgctggt gatggaggta aataagaatt cgatatcaag cttatcgata atcaacctct
     2461 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2521 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2581 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2641 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2701 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2761 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2821 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2881 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2941 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3001 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg
     3061 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3121 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3181 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3241 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3301 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     3361 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3421 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3481 agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3901 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4081 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4141 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4201 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4261 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4321 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4381 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4441 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4501 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4561 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4621 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4681 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4741 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4801 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4861 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4921 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4981 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5041 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5101 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5161 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5221 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5281 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5341 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5401 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5461 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5521 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5581 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5641 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5701 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5761 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5821 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5881 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     5941 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6001 cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
     6061 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     6121 atcactaggg gttccttgta gttaatgatt aacccgccat gctact
//