LOCUS       sequence-366154-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescence reporter fused to EV membrane binding domain for EV 
            imaging and tracking.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4840)
  AUTHORS   Komuro H, Kawai-Harada Y, Aminova S, Pascual N, Malik A, Contag CH, 
            Harada M
  TITLE     Engineering Extracellular Vesicles to Target Pancreatic Tissue In 
            Vivo.
  JOURNAL   Nanotheranostics. 2021 Apr 15;5(4):378-390. doi: 10.7150/ntno.54879.
            eCollection 2021.
  PUBMED    33912378
REFERENCE   2  (bases 1 to 4840)
  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"; doi: 
            "10.7150/ntno.54879"; journalName: "Nanotheranostics"; date: 
            "2021-04-15- 15"; volume: "5"; issue: "4"; pages: "378-390"
COMMENT     Sequence Label: pcS-mCherry-C1C2
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(47..838,839..907))
                     /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     670..689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(908..1012)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1087..1106)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1278..1657
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1658..1861
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1812..1832
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1906..1925
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1906..1924
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2034..2060
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2061..2768
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2206..2224)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2474..2493
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3777..3794
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3817..3834)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3823..4047
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4128..4716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4208..4227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
       61 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      121 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      181 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      241 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      301 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      361 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      421 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      481 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      541 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      601 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      661 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      721 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      781 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      841 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      901 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
      961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1021 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     1081 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     1141 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     1201 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     1261 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     1321 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     1381 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1441 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1501 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1561 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1621 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     1681 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     1741 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     1801 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     1861 tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta
     1921 tagggagacc caagctggct agcgtgtaca aaaaagcagg caccatgccg cgcccccgcc
     1981 tgctggccgc gctgtgcggc gcgctgctct gcgcccccag cctcctcgtc gcctatccat
     2041 atgatgttcc agattatgct atggtgagca agggcgagga ggataacatg gccatcatca
     2101 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
     2161 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
     2221 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     2281 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     2341 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     2401 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     2461 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     2521 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     2581 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     2641 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     2701 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2761 tgtacaaggg tggaggcggt tcaggcggag gtggctctgg cggtggcgga tcggaatgtg
     2821 tcgagccact gggcatggag aatgggaaca ttgccaactc acagatcgcc gcctcatctg
     2881 tgcgtgtgac cttcttgggt ttgcagcatt gggtcccgga gctggcccgc ctgaaccgcg
     2941 caggcatggt caatgcctgg acacccagca gcaatgacga taacccctgg atccaggtga
     3001 acctgctgcg gaggatgtgg gtaacaggtg tggtgacgca gggtgccagc cgcttggcca
     3061 gtcatgagta cctgaaggcc ttcaaggtgg cctacagcct taatggacac gaattcgatt
     3121 tcatccatga tgttaataaa aaacacaagg agtttgtggg taactggaac aaaaacgcgg
     3181 tgcatgtcaa cctgtttgag acccctgtgg aggctcagta cgtgagattg taccccacga
     3241 gctgccacac ggcctgcact ctgcgctttg agctactggg ctgtgagctg aacggatgcg
     3301 ccaatcccct gggcctgaag aataacagca tccctgacaa gcagatcacg gcctccagca
     3361 gctacaagac ctggggcttg catctcttca gctggaaccc ctcctatgca cggctggaca
     3421 agcagggcaa cttcaacgcc tgggttgcgg ggagctacgg taacgatcag tggctgcagg
     3481 tggacctggg ctcctcgaag gaggtgacag gcatcatcac ccagggggcc cgtaactttg
     3541 gctctgtcca gtttgtggca tcctacaagg ttgcctacag taatgacagt gcgaactgga
     3601 ctgagtacca ggaccccagg actggcagca gtaagatctt ccctggcaac tgggacaacc
     3661 actcccacaa gaagaacttg tttgagacgc ccatcctggc tcgctatgtg cgcatcctgc
     3721 ctgtagcctg gcacaaccgc atcgccctgc gcctggagct gctgggctgt accggtcatc
     3781 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     3841 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3901 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3961 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4021 atgctgggga tgcggtgggc tctatggggg gataacgcag gaaagaacat gtgagcaaaa
     4081 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4141 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4201 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4261 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4321 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4381 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4441 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4501 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4561 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4621 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4681 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4741 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4801 aaaaggatct tcacctagat ccttttaaat taaaaatgaa
//
