LOCUS       pLV-puro-EF1A-3XFLAG-hASB8        9430 bp    DNA     circular SYN 18-AUG-2025
DEFINITION  .
ACCESSION   VB200306-1537ajt
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     Promoter        1..229
                     /application_notes="Strong promoter; drives transcription
                     of viral RNA in packaging cells."
                     /description="Rous sarcoma virus enhancer/promoter"
                     /full_name="Rous sarcoma virus (RSV) enhancer/promoter"
                     /note="color: #ef6cdf; direction: RIGHT"
                     /official_designation="RSV"
                     /uuid="3f2ddca7f0cb4015b6abcec8a25c834e"
                     /label="RSV  promoter"
     LTR             230..410
                     /application_notes="Allows transcription of viral RNA and
                     its packaging into virus."
                     /description="Truncated HIV-1 5' long terminal repeat"
                     /full_name="HIV-1 truncated 5' LTR"
                     /note="color: #c54b7c; direction: RIGHT"
                     /official_designation="Delta5' LTR"
                     /uuid="44156a3083604321be44066133e8ecb9"
                     /label="5' LTR-ΔU3"
     domain          1075..1308
                     /application_notes="Rev protein binding site that allows
                     Rev-dependent nuclear export of viral RNA during viral
                     packaging."
                     /description="HIV-1 Rev response element"
                     /full_name="HIV-1 Rev response element"
                     /note="color: #fd3434; direction: RIGHT"
                     /official_designation="RRE"
                     /uuid="df07099e49674cf3aba3661b54e28bd7"
                     /modified_by="u0063854"
                     /label="RRE"
     domain          1803..1920
                     /application_notes="Facilitates the nuclear import of
                     HIV-1 cDNA through a central DNA flap."
                     /description="Central polypurine tract"
                     /full_name="Central polypurine tract"
                     /note="color: #46c6ef; direction: RIGHT"
                     /official_designation="cPPT"
                     /uuid="f7cb29985f9f41a7a5924d3462266f42"
                     /modified_by="u0063854"
                     /label="cPPT"
     Promoter        1959..3137
                     /application_notes="Strong promoter."
                     /description="Human eukaryotic translation elongation
                     factor 1 alpha1 promoter"
                     /full_name="Human eukaryotic translation elongation
                     factor1 alpha1 promoter"
                     /note="color: #eda481; direction: RIGHT"
                     /official_designation="EF1A"
                     /uuid="69c052aac71a4f49bab5b7ae1f46ace8"
                     /label="EF1A"
     CDS             3168..3170
                     /created_by="u0063854"
                     /label
     motif           3171..3236
                     /created_by="User"
                     /modified_by="u0063854"
                     /label="3xFLAG"
     CDS             3237..4103
                     /application_notes="/label=""3xFLAG/hASB8[NM_024095.5]"
                     /note="color: #028a02; direction: RIGHT"
                     /official_designation="hASB8[NM_024095.5]"
                     /uuid="ce7d5db402244d3884b39c391a772808"
                     /modified_by="u0063854"
                     /label="ASB8"
     domain          4142..4739
                     /application_notes="Enhances virus stability in packaging
                     cells, leading to higher titer of packaged virus; enhances
                     higher expression of transgenes."
                     /description="Woodchuck hepatitis virus
                     posttranscriptional regulatory element"
                     /full_name="Woodchuck hepatitis virus posttranscriptional
                     regulatory element"
                     /note="color: #e4b930; direction: RIGHT"
                     /official_designation="WPRE"
                     /uuid="0aedff8308474b369c622433538d7836"
                     /modified_by="u0063854"
                     /label="WPRE"
     Promoter        4758..5267
                     /application_notes="Medium-strength promoter."
                     /description="Mouse phosphoglycerate kinase 1 promoter"
                     /full_name="Mouse phosphoglycerate kinase 1 promoter"
                     /note="color: #7a00b5; direction: RIGHT"
                     /official_designation="mPGK promoter"
                     /uuid="bce93795b5fd4173b8b9abc832edad80"
                     /label="mPGK promoter"
     CDS             5280..5879
                     /application_notes="Allows cells to be resistant to
                     puromycin."
                     /description="Puromycin resistance gene"
                     /full_name="Puromycin resistance gene"
                     /note="color: #05696d; direction: RIGHT"
                     /official_designation="Puro"
                     /uuid="39d7413208494d9291d10b21b13ca44c"
                     /modified_by="u0063854"
                     /label="Puro"
     LTR             5950..6184
                     /application_notes="Allows packaging of viral RNA into
                     virus; self-inactivates the 5' LTR by a copying mechanism
                     during viral genome integration; contains polyadenylation
                     signal for transcription termination."
                     /description="Truncated HIV-1 3' long terminal repeat"
                     /full_name="HIV-1 truncated 3' LTR"
                     /note="color: #f54c6b; direction: RIGHT"
                     /official_designation="DeltaU3/3' LTR"
                     /uuid="c958755414c34372b909e20e81ebcef6"
                     /label="3' LTR-ΔU3"
     PolyA_signal    6257..6391
                     /application_notes="Allows transcription termination and
                     polyadenylation of mRNA transcribed by Pol II RNA
                     polymerase."
                     /description="Simian virus 40 early polyadenylation
                     signal"
                     /full_name="SV40 early polyadenation signal"
                     /note="color: #944603; direction: RIGHT"
                     /official_designation="SV40 early pA"
                     /uuid="2fcb13a62e704d8095b188d9625a6a94"
                     /label="SV40 early pA"
     domain          7345..8205
                     /application_notes="Allows E. coli to be resistant to
                     ampiciIIin."
                     /description="AmpiciIIin resistance gene"
                     /full_name="Ampicillin resistance gene"
                     /note="color: #ce0084; direction: RIGHT"
                     /official_designation="Ampicillin"
                     /uuid="6a7994dc58024d80b36e4a85ad2e1d99"
                     /modified_by="u0063854"
                     /label="AmpiciIIin"
     Rep_origin      8376..8964
                     /application_notes="Facilitates plasmid replication in E.
                     coli; regulates high-copy plasmid number (500-700)."
                     /description="pUC origin of replication"
                     /full_name="pUC origin of replication"
                     /note="color: #6d3ed8; direction: RIGHT"
                     /official_designation="pUC ori"
                     /uuid="64a8cb3def9f49deb6752795c2d893a8"
                     /label="pUC ori"
ORIGIN      
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
     1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     1921 actagtgatt atcggatcaa ctttgtatag aaaagttggg ctccggtgcc cgtcagtggg
     1981 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg
     2041 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2101 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2161 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     2221 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     2281 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     2341 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     2401 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     2461 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     2521 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     2581 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     2641 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg tctcgcgccg ccgtgtatcg
     2701 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     2761 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     2821 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     2881 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     2941 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     3001 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     3061 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     3121 catttcaggt gtcgtgacaa gtttgtacaa aaaagcaggc tgccaccatg gactacaaag
     3181 accatgacgg tgattataaa gatcatgata tcgattacaa ggatgacgat gacaagatga
     3241 gttccagtat gtggtatatt atgcagagca ttcagagcaa atactctctc tccgagcgct
     3301 taatccgaac aattgctgcc atccgttcct tcccacatga taatgtagag gacctcatca
     3361 gagggggagc agatgtgaac tgcactcatg gcacactgaa gcccttgcac tgtgcctgta
     3421 tggtgtcaga tgctgactgt gtggagttac ttctggaaaa aggagccgag gtgaatgccc
     3481 tggatgggta taaccgaaca gccctccact atgcagcaga gaaagatgag gcttgtgtgg
     3541 aggtcctatt ggagtatggt gcaaacccca atgctttgga tggcaacaga gataccccac
     3601 ttcactgggc agcctttaag aacaatgctg agtgtgtgcg ggctctccta gagagcgggg
     3661 cctctgtcaa tgccctggat tacaacaatg atacaccgct cagctgggct gccatgaagg
     3721 gaaatcttga gagtgtcagc atccttctgg attatggcgc agaggtcaga gtcatcaacc
     3781 taataggcca gacacccatc tcccgcctgg tggctctgct agtcagggga cttggaacag
     3841 agaaagagga ctcttgcttt gagctcctcc acagagctgt tggacacttt gaattgagga
     3901 aaaatggcac catgccacga gaggtggcca gagacccgca gctatgtgaa aaactgactg
     3961 ttctgtgctc agctccagga actctaaaaa cactcgctcg ctatgccgtg cgccgtagcc
     4021 tgggactcca gtatctcccc gatgcagtga agggccttcc actgccagct tctttgaagg
     4081 aatacctgtt acttttagaa tagacccagc tttcttgtac aaagtggtga taatcgaatt
     4141 ccgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4201 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4261 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4321 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4381 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4441 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4501 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     4561 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4621 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     4681 gtcttcgcct tcgccctcag acgagtcgga tttcccttcg ggccgcctcc ccgcatcggg
     4741 aattcccgcg gttcgaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     4801 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     4861 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     4921 caccttctac tcctccccta gtcaggaagt tccccccgcc ccgcagctcg cgtcgtgcag
     4981 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
     5041 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
     5101 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
     5161 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
     5221 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctcac gtggccacca
     5281 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     5341 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     5401 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     5461 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     5521 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     5581 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     5641 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     5701 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5761 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5821 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag
     5881 gtacctttaa gaccaatgac ttacaaggca gctgtaaatc ttagccactt tttaaaagaa
     5941 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     6001 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6061 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6121 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6181 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     6241 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6361 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     6421 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6481 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6541 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     6601 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     6661 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     6721 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     6781 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     6841 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     6901 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     6961 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     7021 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     7081 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     7141 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     7201 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     7261 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     7321 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     7381 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     7441 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     7501 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     7561 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     7621 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     7681 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7741 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     7801 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7861 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     7921 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     7981 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     8041 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     8101 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     8161 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     8221 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     8281 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     8341 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     8401 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     8461 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     8521 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     8581 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8641 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8701 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8761 cgtgagctat gagaaagcgc cacgcttccc gaagagagaa aggcggacag gtatccggta
     8821 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8881 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8941 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     9001 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     9061 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     9121 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     9181 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     9241 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     9301 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     9361 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag
     9421 ctgcaagctt
//
