LOCUS       sequence-232790-        9169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V5-tagged Bet1 in lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9169)
  TITLE     Gerace lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9169)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLV-EF1a-V5-Bet1
FEATURES             Location/Qualifiers
     source          1..9169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1..939
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     896..916
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      955..964
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             964..1005
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1531..2080
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1675..1692)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1902..1921
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2083..2682
                     /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(2083..2102)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2579..2599
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2706..3294
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2759..2779)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3177..3188)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3501..4134
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4262..4278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4262..4278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4275..4297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4286..4302
                     /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(4310..4316,4317..4334,4335..4340))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4355..4376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4493..4510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4664..5252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4744..4763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5423..6214,6215..6283))
                     /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     6046..6065
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6284..6388)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6436..6570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6473..6492)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6527..6546
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6738..7371
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7418..7543
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8040..8273
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8458..8502
                     /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    8765..8880
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        8958..9169
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
       61 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
      121 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
      181 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
      241 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
      301 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
      361 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
      421 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
      481 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
      541 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
      601 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
      661 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
      721 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
      781 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
      841 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
      901 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg atccgccgcc
      961 atgggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgggggg cggagggggt
     1021 gggcccatga ggcgtgcagg cctgggtgat ggagcacctc ctggcagcta tgggaactat
     1081 ggctatgcca atactggtta taatgcttgt gaagaagaga atgacagact cactgaaagt
     1141 ctgagaagca aagtaacggc tataaaatct ctttccattg aaataggcca tgaagttaaa
     1201 aatcaaaaca aactattagc tgaaatggat tcacaatttg attctacaac tggatttcta
     1261 ggtaaaacca tgggaagact gaagattttg tccagaggga gccaaacaaa attgctatgc
     1321 tatatgatgc tgttttcatt gtttgtcttt tttgtcattt attggattat taaactgagg
     1381 taatgggccg cgacgatgtt taaacttcga aacgcgtgaa ttcctcgagg gcggccgctc
     1441 tagagtcgac gggccgcggt aacaattgtt aactaactta agctagcaac ggtttccctc
     1501 tagcgggatc aattccgccc ccccccccta acgttactgg ccgaagccgc ttggaataag
     1561 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     1621 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     1681 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     1741 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     1801 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     1861 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     1921 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     1981 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     2041 accacgggga cgtggttttc ctttgaaaaa cacgataata ccatgaccga gtacaagccc
     2101 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     2161 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     2221 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     2281 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     2341 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     2401 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     2461 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     2521 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     2581 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     2641 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacccgggcg gggcgcgtct
     2701 ggaacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2761 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2821 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2881 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2941 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3001 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3061 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     3121 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3181 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3241 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     3301 ttaattctgc agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
     3361 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
     3421 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     3481 tttaaaagaa aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct
     3541 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
     3601 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
     3661 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
     3721 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
     3781 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
     3841 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
     3901 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
     3961 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     4021 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     4081 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     4141 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     4201 agaggccttg acattgctag cgtttaccgt cgacctctag ctagagcttg gcgtaatcat
     4261 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4321 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4381 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4441 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4501 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4561 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4621 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4681 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4741 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4801 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4861 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4921 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4981 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5041 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5101 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5161 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5221 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5281 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5341 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5401 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5461 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5521 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5581 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5641 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5701 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5761 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5821 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5881 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5941 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6001 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6061 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6121 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6181 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6241 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6301 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6361 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     6421 acggatcggg agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6481 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6541 ctcatcaatg tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat
     6601 cccaaacttc ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc
     6661 taaacctgtg attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt
     6721 actacaaact tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga
     6781 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     6841 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     6901 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     6961 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     7021 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     7081 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     7141 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     7201 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7261 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7321 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
     7381 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
     7441 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     7501 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     7561 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     7621 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     7681 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     7741 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     7801 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     7861 taagaccacc gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg
     7921 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     7981 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     8041 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     8101 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     8161 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     8221 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     8281 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     8341 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     8401 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     8461 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     8521 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     8581 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     8641 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     8701 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat
     8761 cgcctttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     8821 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     8881 tcgggtttat tacagggaca gcagagatcc agtttatcga tgaggccctt tcgtcttcac
     8941 tcgaggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     9001 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     9061 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     9121 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacag
//
