LOCUS       sequence-251160-        8079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector for expression of HA-Tagged E6-AP I AA 1-87 in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8079)
  AUTHORS   Kuhnle S, Martinez-Noel G, Leclere F, Hayes SD, Harper JW, Howley PM
  TITLE     Angelman syndrome-associated point mutations in the Zn-binding 
            N-terminal (AZUL) domain of UBE3A ubiquitin ligase inhibit binding 
            to the proteasome.
  JOURNAL   J Biol Chem. 2018 Sep 26. pii: RA118.004653. doi: 
            10.1074/jbc.RA118.004653.
  PUBMED    30257870
REFERENCE   2  (bases 1 to 8079)
  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 Biol 
            Chem. 2018 Sep 26. pii: RA118.004653. doi: 
            10.1074/jbc.RA118.004653."
COMMENT     Sequence Label: PHAGE-P CMVt N-HA hE6-AP II (AA 1-87)
FEATURES             Location/Qualifiers
     source          1..8079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2227..2530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2531..2734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2685..2705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2736..2754
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2757..2775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2785..2809
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2896..2905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2908..2934
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2950..2974
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3240..3264)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3283..3782
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3318..3341)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3715..3734
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3803..4402
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3803..3822)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4299..4319
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4469..5057
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4522..4542)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4940..4951)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5132..5365
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5484..5502)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5570..5674
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5675..5743,5744..6535)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5893..5912)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6706..7294
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7195..7214
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7448..7465
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7705..7726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7741..7746,7747..7764,7765..7771)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7779..7795
                     /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     7784..7806
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7803..7819
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7803..7819
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7869..7888
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata gagatctccc
     2761 tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg
     2821 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
     2881 agcgtttaaa cgctagccac catggcgtac ccatacgatg ttccagatta cgctgctcga
     2941 ggttatcaaa caagtttgta caaaaaagtt ggcaccatgg agaagctgca ccagtgttat
     3001 tggaaatcag gagaacctca gtctgacgac attgaagcta gccgaatgaa gcgagcagct
     3061 gcaaagcatc taatagaacg ctactaccac cagttaactg agggctgtgg aaatgaagcc
     3121 tgcacgaatg agttttgtgc ttcctgtcca acttttcttc gtatggataa taatgcagca
     3181 gctattaaag ccctcgagct ttataagatt aatgcaaaac tctgtgatcc tcatccctga
     3241 cccaactttc ttgtacaaag tggttcgata acgaattcta ccgggtaggg gaggcgcttt
     3301 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
     3361 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
     3421 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
     3481 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc
     3541 agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc
     3601 agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc
     3661 tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
     3721 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
     3781 cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3841 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3901 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3961 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4021 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4081 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4141 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4201 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4261 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     4321 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     4381 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     4441 cgcacgaccc catgcatcga tagatcctaa tcaacctctg gattacaaaa tttgtgaaag
     4501 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     4561 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     4621 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     4681 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     4741 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     4801 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     4861 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     4921 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     4981 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5041 ttgggccgcc tccccgcctg agatccttta agaccaatga cttacaaggc agctgtagat
     5101 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     5161 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     5221 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     5281 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     5341 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     5401 taacttgcaa agaaatgaat atcagagagt gagaggcccg ggttaattaa ggaaagggct
     5461 agatcattct tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5521 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5581 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5641 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5701 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5761 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5821 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5881 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     5941 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6001 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6061 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6121 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6181 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6241 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6301 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6361 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6421 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6481 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6541 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     6601 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     6661 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6721 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6781 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6841 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6901 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6961 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7021 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7081 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7141 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7201 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7261 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7321 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7381 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     7441 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     7501 ccccgcgcgt tggccgattc attaatgcag caagctcatg gctgactaat tttttttatt
     7561 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     7621 tttggaggcc taggcttttg caaaaagctc cccgtggcac gacaggtttc ccgactggaa
     7681 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     7741 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7801 cacaggaaac agctatgaca tgattacgaa tttcacaaat aaagcatttt tttcactgca
     7861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
     7921 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
     7981 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
     8041 aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
