LOCUS       sequence-384501-        5877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CreiLOV with FusionRed tag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5877)
  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 5877)
  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-CreiLOV-P2A-FusionRed
FEATURES             Location/Qualifiers
     source          1..5877
                     /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             1366..1422
                     /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             1435..2133
                     /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    2158..2746
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2211..2231)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2629..2640)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2789..2910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2800..2819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2854..2873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3441..4029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3521..3540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4200..4991,4992..5060))
                     /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     4823..4842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5061..5165)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5191..5646)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5328..5349)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5540..5559
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5718..5847
                     /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 rggsggggsg
      421 ggggggggsg rggggsgggg sggggsgrgg 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 caccatggcc ggcctgagac acaccttcgt ggtggccgac
     1021 gccaccctgc ccgactgccc cctggtgtac gccagcgagg gcttctacgc catgaccggc
     1081 tacggccccg acgaggtgct gggccacaac gccagattcc tgcagggcga gggcaccgac
     1141 cccaaggagg tgcagaagat cagagacgcc atcaagaagg gcgaggcctg cagcgtgaga
     1201 ctgctgaact acagaaagga cggcaccccc ttctggaacc tgctgaccgt gacccccatc
     1261 aagacccccg acggcagagt gagcaagttc gtgggcgtgc aggtggacgt gaccagcaag
     1321 accgagggca aggccctggc ccgaccggtt cacgtgggaa gtggggctac aaacttctca
     1381 ctgctgaaac aggccgggga tgtggaggaa aacccaggac cacacgtgac tagtatggtg
     1441 agcgagctga ttaaggagaa catgcccatg aagctgtaca tggagggcac cgtgaacaac
     1501 caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
     1561 agaatcaagg tcgtcgaggg cggccctctc cccttcgcct tcgacatcct ggctaccagc
     1621 ttcatgtacg gcagcagaac cttcatcaag caccctccgg gcatccccga cttctttaag
     1681 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg
     1741 ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaaggtt
     1801 agaggggtga acttcccagc caacggccct gtgatgcaga agaaaacact cggctgggag
     1861 gcctccaccg agacgatgta ccctgctgac ggaggactgg aaggcgcatg tgacatggct
     1921 ctgaagctcg tgggaggagg acacctgatc tgcaaccttg agacaacata cagatccaag
     1981 aaacccgcta cgaacctcaa gatgcccggc gtctacaacg tggaccacag actggaaaga
     2041 atcaaggagg ccgacgatga gacatacgtc gagcagcacg aggtggctgt ggccagatac
     2101 tctactggtg gcgctggtga tggaggtaaa taagaattcg atatcaagct tatcgataat
     2161 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2221 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2281 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2341 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     2401 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     2461 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2521 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     2581 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     2641 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     2701 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
     2761 cccgggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     2821 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2881 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2941 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     3001 aaaatcgata aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat
     3061 cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3121 gctcactgac tcgctgcgct cggtcgtkcg gcwrcrkcga kmrgkatcwt cctagagcat
     3181 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     3241 gagttggcca ctccctctct gcgcgctcgc tcgctcactg actcgctgcg ctcggtcgtt
     3301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5161 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     5221 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     5281 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     5341 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     5401 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     5461 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     5521 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     5581 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     5641 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     5701 cttcgctatt acgccagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
     5761 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
     5821 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctact
//
