LOCUS       sequence-389471-        5698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express miniGFP2 and FusionRed in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5698)
  AUTHORS   Liang GT, Lai C, Yue Z, Zhang H, Li D, Chen Z, Lu X, Tao L, Subach 
            FV, Piatkevich KD
  TITLE     Enhanced small green fluorescent proteins as a multisensing platform
            for biosensor development.
  JOURNAL   Front Bioeng Biotechnol. 2022 Oct 17;10:1039317. doi: 
            10.3389/fbioe.2022.1039317. eCollection 2022.
  PUBMED    36324888
REFERENCE   2  (bases 1 to 5698)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Bioeng Biotechnol."; date: "2022-10-17"; volume: "10:1039317. doi"; 
            pages: " 10.3389/fbioe.2022.1039317. eCollection 2022"
COMMENT     Sequence Label: pAAV-CAG-miniGFP2-P2A-FusionRed
FEATURES             Location/Qualifiers
     source          1..5698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        281..558
                     /label=chicken beta-actin promoter
     primer_bind     837..859
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     921..940
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      979..988
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1339..1395
                     /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             1408..2106
                     /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    2131..2719
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2184..2204)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2602..2613)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2762..2883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2773..2792)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2827..2846
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3262..3850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3342..3361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4021..4812,4813..4881))
                     /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     4644..4663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4882..4986)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5012..5467)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5149..5170)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5361..5380
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5539..5668
                     /label=AAV2 ITR
ORIGIN
        1 tatctacgta gccatgctct aggaagagta ccattgacgt caataatgac gtatgttccc
       61 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      121 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      181 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      241 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      301 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      361 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
      421 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
      481 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      541 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      601 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      661 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      721 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
      781 ggctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
      841 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
      901 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
      961 agaattggat ccggtaccgc caccatggag aagtcctttg tgattacaga cccatggctg
     1021 cccgactacc ctattatttc cgctagtgac ggattcctgg agctgaccga gtattctcgg
     1081 gatgaaatca tgggaaggaa cgcacgcttc ctgcagggac ccgagactga ccaggccacc
     1141 gtgcagaaga tccgggacgc tattcgagat cggagaccta ccacagtcca gctgatcaac
     1201 tacacaaaga gcggcaagaa gttctggaat ctgttgcacc tgcagcctgt gtttgacgga
     1261 aaaggcgggc tgcagtattt tattggagtg cagctggtcg gctccgatca tgtccgaccg
     1321 gttcacgtgg gaagtggggc tacaaacttc tcactgctga aacaggccgg ggatgtggag
     1381 gaaaacccag gaccacacgt gactagtatg gtgagcgagc tgattaagga gaacatgccc
     1441 atgaagctgt acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc
     1501 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtcgtcga gggcggccct
     1561 ctccccttcg ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc
     1621 aagcaccctc cgggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg
     1681 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
     1741 caggacggct gcctcatcta caacgtcaag gttagagggg tgaacttccc agccaacggc
     1801 cctgtgatgc agaagaaaac actcggctgg gaggcctcca ccgagacgat gtaccctgct
     1861 gacggaggac tggaaggcgc atgtgacatg gctctgaagc tcgtgggagg aggacacctg
     1921 atctgcaacc ttgagacaac atacagatcc aagaaacccg ctacgaacct caagatgccc
     1981 ggcgtctaca acgtggacca cagactggaa agaatcaagg aggccgacga tgagacatac
     2041 gtcgagcagc acgaggtggc tgtggccaga tactctactg gtggcgctgg tgatggaggt
     2101 aaataagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     2161 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2221 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2281 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2341 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2401 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2461 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2521 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2581 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2641 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2701 ctttgggccg cctccccgca tcgataccgt cgacccgggc ggccgcttcg agcagacatg
     2761 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2821 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2881 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     2941 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     3001 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     3061 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gactcgctgc gctcggtcgt
     3121 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3181 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3241 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3301 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3361 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3601 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3661 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3721 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3781 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4141 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4981 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5041 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5101 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5161 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5221 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5281 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5341 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     5401 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     5461 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     5521 tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc
     5581 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
     5641 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctact
//
