LOCUS       sequence-453499-        7540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  secretory SortaseA in p53 (SRE backbone).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7540)
  AUTHORS   Gonzalez-Callejo P, Guo Z, Ziglari T, Claudio NM, Nguyen KH, 
            Oshimori N, Seras-Franzoso J, Pucci F
  TITLE     Cancer stem cell-derived extracellular vesicles preferentially 
            target MHC-II-macrophages and PD1+ T cells in the tumor 
            microenvironment.
  JOURNAL   PLoS One. 2023 Feb 3;18(2):e0279400. doi: 
            10.1371/journal.pone.0279400. eCollection 2023.
  PUBMED    36735677
REFERENCE   2  (bases 1 to 7540)
  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: "PLoS One.";
            date: "2023-02-3"; volume: "18(2):e0279400. doi"; pages: " 
            10.1371/journal.pone.0279400. eCollection 2023"
COMMENT     Sequence Label: P55.SRE.mCMV.flag.mSrtA-3M.wpre
FEATURES             Location/Qualifiers
     source          1..7540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     42..61
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      112..247
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(267..286)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(268..286)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(296..313)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(296..312)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(305..327)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      454..909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(541..560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     751..772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        935..1039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1040..1108,1109..1900)
                     /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     complement(1258..1277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2071..2659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2560..2579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2813..2830
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2947..2968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2983..2988,2989..3006,3007..3013)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3021..3037
                     /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)."
     primer_bind     3026..3048
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3045..3061
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3045..3061
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3080..3100
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3082..3100
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        3201..3580
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3581..3784
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3735..3755
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3799..3979
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4026..4151
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4644..4877
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5062..5106
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5373..5490
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      5793..5802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5862..5885
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(6608..6628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7026..7037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7186..7419
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    join(7491..7540,1..72)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(7528..7540,1..7))
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
       61 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
      121 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
      181 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
      241 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
      301 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      361 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      421 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
      481 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
      541 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
      601 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
      661 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
      721 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
      781 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      841 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
      901 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      961 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1021 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1081 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1141 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     1201 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     1261 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     1321 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1381 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1441 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1501 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1561 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1621 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1681 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1741 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1801 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1861 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1921 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1981 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     2041 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2101 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2161 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2221 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2281 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2341 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2401 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2461 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2521 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2581 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2641 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2701 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2761 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2821 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2881 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2941 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     3001 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     3061 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
     3121 agcttggcca ttgcatacgt tgtatccata tcataatatg tacatttata ttggctcatg
     3181 tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
     3241 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3301 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3361 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3421 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     3481 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     3541 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3601 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3661 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     3721 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     3781 agctcgttta gtgaaccggg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     3841 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     3901 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     3961 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
     4021 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
     4081 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
     4141 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
     4201 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
     4261 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
     4321 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
     4381 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
     4441 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc
     4501 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     4561 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     4621 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     4681 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     4741 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     4801 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     4861 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     4921 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     4981 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     5041 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     5101 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     5161 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     5221 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     5281 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     5341 ttagtgaacg gatctcgacg gtatcggtta acttttaaaa gaaaaggggg gattgggggg
     5401 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     5461 caaaaacaaa ttacaaaaat tcaaaatttt atcgatcacg agactagcct cgagagccag
     5521 acagggtaag ccagacatca gcagccagac aatcagagcc agacacattc agccagacat
     5581 aacgagccag acagtccaag ccagacaacg ttagccagac atttgtagcc agacattgaa
     5641 agccagacag aaggagccag acatactgag ccagacagta cggtgggagg cctatataag
     5701 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     5761 ccatagaaga caccgggacc gatccaccgg tcgccaccat ggagactgac acattgcttc
     5821 tgtgggtcct ccttctttgg gttccaggaa gcactggcga cgactacaag gacgacgatg
     5881 ataaagggtc cggcctggaa caagctaaac cccaaattcc caaggataaa agcaaggtag
     5941 ctggatacat agaaatacct gatgccgata ttaaggaacc cgtttacccc ggtccagcta
     6001 cttctgaaca actgaatcgc ggagtctctt tcgcagaaga gaacgagtcc ttggatgacc
     6061 aaaacatttc aatcgcaggt cacactttta tagataggcc aaactaccaa tttactaatc
     6121 tgaaagctgc taagaagggt tctatggtct actttaaggt agggaacgag acccggaagt
     6181 acaaaatgac cagtattcgc aatgtgaagc ccacagatgt gggtgtactt gatgaacaaa
     6241 agggaaaaga caaacagttg acactgataa cttgcgacga ttacaacgag aagactggag
     6301 tttgggagac acgaaagatc tttgtagcaa cagaggtcaa aggtagtacc agcggatctg
     6361 gcaagcccgg atccggcgag ggcagtacaa aaggtaatgc cgtgggtcaa gacacacaag
     6421 aggttattgt cgtgccccac agtcttccat tcaaggttgt cgtcattagt gccatcttgg
     6481 cccttgtcgt cctcactatt atctccctga taatattgat aatgctctgg cagaagaaac
     6541 ctcgctaggt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     6601 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     6661 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     6721 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     6781 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     6841 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     6901 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     6961 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     7021 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctagagc
     7081 ctcttccgcg tcttcgcctt cccgggtcga gctcggtacc tttaagacca atgacttaca
     7141 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     7201 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     7261 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     7321 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     7381 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     7441 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     7501 ttgcagctta taatggttac aaataaagca atagcatcac
//
