LOCUS       sequence-331262-        7236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  human CD81-TEV cleavage site-Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  AUTHORS   Somiya M, Kuroda S
  TITLE     Reporter gene assay for membrane fusion of extracellular vesicles.
  JOURNAL   J Extracell Vesicles. 2021 Nov;10(13):e12171. doi: 
            10.1002/jev2.12171.
  PUBMED    34807503
REFERENCE   2  (bases 1 to 7236)
  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: "J Extracell
            Vesicles."; date: "2021-11"; volume: "10(13):e12171. doi"; pages: " 
            10.1002/jev2.12171"
COMMENT     Sequence Label: pcDNA3.1-CD81-(TEV)-Cre
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        241..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     869..888
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        869..887
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      992..1001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1706..1726
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1730..2752
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS
                     VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR
                     PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG
                     IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW
                     ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY
                     LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG
                     D"
     primer_bind     complement(2833..2850)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2839..3063
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3109..3537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3196..3215)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3406..3427
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3546..3566)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3551..3880
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3731..3866
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3793..3812
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3947..4741
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4001..4020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4611..4630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4915..5036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4952..4971)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5006..5025
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5085..5101)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5085..5101)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5098..5120)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5109..5125
                     /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(5133..5139,5140..5157,5158..5163))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5178..5199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5316..5333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5487..6075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5567..5586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6246..7037,7038..7106))
                     /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     6869..6888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7107..7211)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
       61 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
      121 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      181 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      841 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      901 gctagcgttt aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa
      961 ttctgcagat atccagcaca gtggcggccg cgccaccatg ggagtggagg gctgcaccaa
     1021 gtgcatcaag tacctgctct tcgtcttcaa tttcgtcttc tggctggctg gaggcgtgat
     1081 cctgggtgtg gccctgtggc tccgccatga cccgcagacc accaacctcc tgtatctgga
     1141 gctgggagac aagcccgcgc ccaacacctt ctatgtaggc atctacatcc tcatcgctgt
     1201 gggcgctgtc atgatgttcg ttggcttcct gggctgctac ggggccatcc aggaatccca
     1261 gtgcctgctg gggacgttct tcacctgcct ggtcatcctg tttgcctgtg aggtggccgc
     1321 cggcatctgg ggctttgtca acaaggacca gatcgccaag gatgtgaagc agttctatga
     1381 ccaggcccta cagcaggccg tggtggatga tgacgccaac aacgccaagg ctgtggtgaa
     1441 gaccttccac gagacgcttg actgctgtgg ctccagcaca ctgactgctt tgaccacctc
     1501 agtgctcaag aacaatttgt gtccctcggg cagcaacatc atcagcaacc tcttcaagga
     1561 ggactgccac cagaagatcg atgacctctt ctccgggaag ctgtacctca tcggcattgc
     1621 tgccatcgtg gtcgctgtga tcatgatctt cgagatgatc ctgagcatgg tgctgtgctg
     1681 tggcatccgg aacagctccg tgtacgagaa cctgtacttc cagtctgcca atttactgac
     1741 cgtacaccaa aatttgcctg cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa
     1801 cctgatggac atgttcaggg atcgccaggc gttttctgag catacctgga aaatgcttct
     1861 gtccgtttgc cggtcgtggg cggcatggtg caagttgaat aaccggaaat ggtttcccgc
     1921 agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc tggcagtaaa
     1981 aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg ggctgccacg
     2041 accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag aaaacgttga
     2101 tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg accaggttcg
     2161 ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat ttctggggat
     2221 tgcttataac accctgttac gtatagccga aattgccagg atcagggtta aagatatctc
     2281 acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc tggttagcac
     2341 cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc gatggatttc
     2401 cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca gaaaaaatgg
     2461 tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag ggatttttga
     2521 agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat acctggcctg
     2581 gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg gagtttcaat
     2641 accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga actatatccg
     2701 taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg atggaccggt
     2761 ggaacaaaaa ctttatttct gaagaagatc tgtgactcga gtctagaggg cccgtttaaa
     2821 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2881 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2941 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3001 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3061 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3121 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3181 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3241 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3301 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3361 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3421 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3481 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3541 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3601 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3661 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3721 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3781 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3841 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3901 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3961 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4021 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4081 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4141 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4201 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4261 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4321 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4381 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4441 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4501 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4561 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4621 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4681 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4741 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4801 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4861 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     4921 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4981 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     5041 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     5101 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     5161 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     5221 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5281 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5341 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5401 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5461 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5521 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5581 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5641 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5701 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5761 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5821 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5881 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5941 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6001 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6061 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6121 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6181 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6241 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6301 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6361 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6421 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6481 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6541 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6601 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6661 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6721 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6781 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6841 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6901 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6961 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7021 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7081 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7141 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7201 ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
