LOCUS       sequence-356147-        6569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bioluminescence 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 6569)
  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 6569)
  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: pcDNA-gLuc-C1C2
FEATURES             Location/Qualifiers
     source          1..6569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(542..1333,1334..1402))
                     /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     1165..1184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1403..1507)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1582..1601)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1773..2152
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2153..2356
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2307..2327
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2401..2420
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2401..2419
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2529..2555
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4023..4040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4063..4080)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4069..4293
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4339..4767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4426..4445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4636..4657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4776..4796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4781..5110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4961..5096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5023..5042
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5158..5205
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5224..5622
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5780..5901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5817..5836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5871..5890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5950..5966)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5950..5966)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5963..5985)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5974..5990
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5998..6004,6005..6022,6023..6028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6043..6064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6181..6198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6352..6569,1..371))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6432..6451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
       61 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
      121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
      181 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
      241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
      301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
      361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
      421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
      481 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
      541 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
      601 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      661 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
      721 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
      781 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
      841 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
      901 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
      961 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     1021 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     1081 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     1141 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     1201 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     1261 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     1321 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     1381 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     1441 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     1501 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     1561 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     1621 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     1681 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     1741 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     1801 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     1861 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     1921 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     1981 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2041 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2101 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     2161 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     2221 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2281 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2341 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     2401 taatacgact cactataggg agacccaagc tggctagcgt gtacaaaaaa gcaggcacca
     2461 tgccgcgccc ccgcctgctg gccgcgctgt gcggcgcgct gctctgcgcc cccagcctcc
     2521 tcgtcgccta tccatatgat gttccagatt atgctaagcc caccgagaac aacgaagact
     2581 tcaacatcgt ggccgtggcc agcaacttcg cgaccacgga tctcgatgct gaccgcggga
     2641 agttgcccgg caagaagctg ccgctggagg tgctcaaaga gatggaagcc aatgcccgga
     2701 aagctggctg caccaggggc tgtctgatct gcctgtccca catcaagtgc acgcccaaga
     2761 tgaagaagtt catcccagga cgctgccaca cctacgaagg cgacaaagag tccgcacagg
     2821 gcggcatagg cgaggcgatc gtcgacattc ctgagattcc tgggttcaag gacttggagc
     2881 ccatggagca gttcatcgca caggtcgatc tgtgtgtgga ctgcacaact ggctgcctca
     2941 aagggcttgc caacgtgcag tgttctgacc tgctcaagaa gtggctgccg caacgctgtg
     3001 cgacctttgc cagcaagatc cagggccagg tggacaagat caagggggcc ggtggtgaca
     3061 aatgtgtcga gccactgggc atggagaatg ggaacattgc caactcacag atcgccgcct
     3121 catctgtgcg tgtgaccttc ttgggtttgc agcattgggt cccggagctg gcccgcctga
     3181 accgcgcagg catggtcaat gcctggacac ccagcagcaa tgacgataac ccctggatcc
     3241 aggtgaacct gctgcggagg atgtgggtaa caggtgtggt gacgcagggt gccagccgct
     3301 tggccagtca tgagtacctg aaggccttca aggtggccta cagccttaat ggacacgaat
     3361 tcgatttcat ccatgatgtt aataaaaaac acaaggagtt tgtgggtaac tggaacaaaa
     3421 acgcggtgca tgtcaacctg tttgagaccc ctgtggaggc tcagtacgtg agattgtacc
     3481 ccacgagctg ccacacggcc tgcactctgc gctttgagct actgggctgt gagctgaacg
     3541 gatgcgccaa tcccctgggc ctgaagaata acagcatccc tgacaagcag atcacggcct
     3601 ccagcagcta caagacctgg ggcttgcatc tcttcagctg gaacccctcc tatgcacggc
     3661 tggacaagca gggcaacttc aacgcctggg ttgcggggag ctacggtaac gatcagtggc
     3721 tgcaggtgga cctgggctcc tcgaaggagg tgacaggcat catcacccag ggggcccgta
     3781 actttggctc tgtccagttt gtggcatcct acaaggttgc ctacagtaat gacagtgcga
     3841 actggactga gtaccaggac cccaggactg gcagcagtaa gatcttccct ggcaactggg
     3901 acaaccactc ccacaagaag aacttgtttg agacgcccat cctggctcgc tatgtgcgca
     3961 tcctgcctgt agcctggcac aaccgcatcg ccctgcgcct ggagctgctg ggctgtaccg
     4021 gtcatcatca ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta
     4081 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     4141 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     4201 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     4261 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     4321 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     4741 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4801 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4861 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4921 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4981 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5041 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5101 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     5161 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     5221 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     5281 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     5341 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     5401 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     5461 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     5521 ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg
     5581 attcgtgaat tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga
     5641 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5701 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5761 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5881 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5941 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6001 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6061 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6121 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6181 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6241 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6301 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6361 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6421 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6481 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6541 ttctcatagc tcacgctgta ggtatctca
//
