LOCUS       pLV-puro-EF1A-3XFLAG-hASB8_L199G        9431 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"
     motif           1074..1307
                     /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"
     motif           1802..1919
                     /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        1958..3136
                     /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             3167..3169
                     /created_by="u0063854"
                     /label
     motif           3170..3235
                     /created_by="User"
                     /modified_by="u0063854"
                     /label="3xFLAG"
     CDS             3236..4102
                     /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"
     conflict        3830..3832
                     /created_by="u0063854"
                     /label="L199G"
     motif           4141..4738
                     /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        4757..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..6185
                     /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    6258..6392
                     /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"
     ORF             7346..8206
                     /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"
                     /label="AmpiciIIin"
     Rep_origin      8377..8965
                     /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 gcggggagaa
      601 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      661 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      721 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      781 cagaagaact taaatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
      841 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
      901 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
      961 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1021 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1081 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1141 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1201 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1261 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1321 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1381 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1441 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1501 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     1561 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     1621 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     1681 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     1741 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgctag
     1801 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1861 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta
     1921 ctagtgatta tcggatcaac tttgtataga aaagttgggc tccggtgccc gtcagtgggc
     1981 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg
     2041 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     2101 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     2161 tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg
     2221 cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt
     2281 gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta
     2341 aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt
     2401 gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta
     2461 aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg
     2521 ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc
     2581 gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg
     2641 ggggtagtct caagctggcc ggcctgctct ggtgcctggt ctcgcgccgc cgtgtatcgc
     2701 cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc
     2761 gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc
     2821 gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga
     2881 ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac
     2941 gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt
     3001 ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt
     3061 tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc
     3121 atttcaggtg tcgtgacaag tttgtacaaa aaagcaggct gccaccatgg actacaaaga
     3181 ccatgacggt gattataaag atcatgatat cgattacaag gatgacgatg acaagatgag
     3241 ttccagtatg tggtatatta tgcagagcat tcagagcaaa tactctctct ccgagcgctt
     3301 aatccgaaca attgctgcca tccgttcctt cccacatgat aatgtagagg acctcatcag
     3361 agggggagca gatgtgaact gcactcatgg cacactgaag cccttgcact gtgcctgtat
     3421 ggtgtcagat gctgactgtg tggagttact tctggaaaaa ggagccgagg tgaatgccct
     3481 ggatgggtat aaccgaacag ccctccacta tgcagcagag aaagatgagg cttgtgtgga
     3541 ggtcctattg gagtatggtg caaaccccaa tgctttggat ggcaacagag ataccccact
     3601 tcactgggca gcctttaaga acaatgctga gtgtgtgcgg gctctcctag agagcggggc
     3661 ctctgtcaat gccctggatt acaacaatga tacaccgctc agctgggctg ccatgaaggg
     3721 aaatcttgag agtgtcagca tccttctgga ttatggcgca gaggtcagag tcatcaacct
     3781 aataggccag acacccatct cccgcctggt ggctctgcta gtcaggggag gtggaacaga
     3841 gaaagaggac tcttgctttg agctcctcca cagagctgtt ggacactttg aattgaggaa
     3901 aaatggcacc atgccacgag aggtggccag agacccgcag ctatgtgaaa aactgactgt
     3961 tctgtgctca gctccaggaa ctctaaaaac actcgctcgc tatgccgtgc gccgtagcct
     4021 gggactccag tatctccccg atgcagtgaa gggccttcca ctgccagctt ctttgaagga
     4081 atacctgtta cttttagaat agacccagct ttcttgtaca aagtggtgat aatcgaattc
     4141 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     4201 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4261 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4321 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4381 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4441 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4501 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     4561 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4621 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4681 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcggga
     4741 attcccgcgg ttcgaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc
     4801 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac
     4861 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc
     4921 accttctact cctcccctag tcaggaagtt cccccccgcc 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 gagaatccct cagacccttt tagtcagtgt ggaaaatctc
     6181 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     6241 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6301 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6361 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     6421 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     6481 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     6541 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     6601 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     6661 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     6721 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     6781 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6841 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6901 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6961 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7021 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7081 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     7141 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     7201 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     7261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     7321 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     7381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     7441 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     7501 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     7561 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     7621 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     7681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     7741 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     7801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     7861 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     7921 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     7981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     8041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     8101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     8161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     8221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     8401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     8461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     8521 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     8581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     8641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     8701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagagaga aaggcggaca ggtatccggt
     8821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     8941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     9001 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     9061 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     9121 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     9181 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     9241 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     9301 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     9361 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
     9421 gctgcaagct t
//
