LOCUS       sequence-232781-        7223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V5-tagged Itprip in lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7223)
  AUTHORS   Cheng LC, Baboo S, Lindsay C, Brusman L, Martinez-Bartolome S, Tapia
            O, Zhang X, Yates JR 3rd, Gerace L
  TITLE     Identification of new transmembrane proteins concentrated at the 
            nuclear envelope using organellar proteomics of mesenchymal cells.
  JOURNAL   Nucleus. 2019 Dec;10(1):126-143. doi: 10.1080/19491034.2019.1618175.
  PUBMED    31142202
REFERENCE   2  (bases 1 to 7223)
  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"; doi: 
            "10.1080/19491034.2019"; journalName: "Nucleus"; date: "2019-12"; 
            volume: "10"; issue: "1"; pages: "126-143"
COMMENT     Sequence Label: pLV-EF1a-Itprip-V5
FEATURES             Location/Qualifiers
     source          1..7223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          213..1151
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1108..1128
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1182..1191
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2856..2897
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3034..3583
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3178..3195)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3405..3424
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3586..4185
                     /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(3586..3605)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4082..4102
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4209..4797
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4262..4282)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4680..4691)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5004..5637
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5684..5809
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6306..6539
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6724..6768
                     /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    7031..7146
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
ORIGIN
        1 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
       61 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
      121 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
      181 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
      241 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
      301 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
      361 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
      421 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
      481 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
      541 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
      601 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
      661 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
      721 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
      781 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
      841 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
      901 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
      961 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     1021 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     1081 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     1141 ttccatttca ggtgtcgtga ggatccgcca tcggcccgcc cgccgccatg gccatggggc
     1201 tcttccgcgt gtgtctggtg gtggtgacgg ccatcatcaa ccacccgctg ctgttcccgc
     1261 gggagaacgc cacagtcccc gagaacgagg aggagatcat ccgcaagatg caggcgcacc
     1321 aggagaagct gcagctggag cagttgcgcc tggaggagga ggtggctcgg ctggcggccg
     1381 aaaaggaggc actggagcag gtggcggagg agggcaggca gcagaacgag acacgcgtgg
     1441 cctgggacct ctggagcacc ctctgcatga tcctcttcct gatgatcgag gtgtggcggc
     1501 aggaccacca ggaggggccc tcacctgagt gcctgggcgg tgaggaggat gagctgcctg
     1561 ggctgggggg cgcccccttg cagggcctca ccctgcccaa caaggccacg cttggccact
     1621 tttatgagcg ctgcatccgg ggggccacgg ccgatgcagc ccgtacccgg gagttcctgg
     1681 aaggcttcgt ggatgacttg ctggaagccc tgaggagcct ctgcaaccgg gacaccgaca
     1741 tggaggtgga ggacttcatt ggcgtggaca gcatgtacga gaactggcag gtggacaggc
     1801 cactgctgtg ccaccttttc gtgcccttca caccccccga gccctaccgc ttccacccag
     1861 agctctggtg ctccggccgc tcagtgcccc tggatcgcca gggctacggc cagatcaagg
     1921 tggtccgcgc cgatggggac acattgagct gcatctgcgg caagaccaag ctcggggaag
     1981 acatgctgtg tctcctgcac ggcaggaaca gcatggcgcc tccctgcggc gacatggaga
     2041 acctgctgtg tgccacagat tccctgtacc tggacacgat gcaggtcatg aagtggttcc
     2101 agacggccct caccagagcc tggaagggca tcgcccacaa gtacgagttc gacctggcct
     2161 ttggccagct ggacagcccg gggtccctga agatcaagtt ccgttcaggg aagttcatgc
     2221 ccttcaacct gattcctgtg atccagtgtg atgactcgga cctgtacttt gtctcccacc
     2281 ttcccaggga gccctctgag ggcaccccag cctccagcac agactggctc ctgtcctttg
     2341 ctgtctatga gcgacacttc ctcaggacga cactaaaggc actgcccgag ggcgcctgcc
     2401 acctcagctg cctgcagata gcatccttcc tgctctccaa gcagagccgc ctgaccggtc
     2461 ccagcgggct cagcagctac cacctgaaga cggccctact gcacctccta ctcctccggc
     2521 aggccgccga ctggaaggcg gggcagctgg acgctcgtct gcacgagttg ctgtgcttcc
     2581 tggagaagag cttgctccag aagaagctcc accacttctt catcggcaac cgcaaggtgc
     2641 ctgaggccat gggactccct gaggccgtgc tcagggccga gcccctcaac ctcttccggc
     2701 ccttcgtcct gcagcgaagc ctttaccgta agacactgga ctccttctat gagatgctca
     2761 agaatgcccc agcgctcatt agcgagtatt ccctacatgt cccctcagac cagcctaccc
     2821 caaaaagcgg tgggccgggc gggggtggcg ggggaggtaa gcctatccct aaccctctcc
     2881 tcggtctcga ttctacgtga gtttaaactt cgaaacgcgt gaattcctcg agggcggccg
     2941 ctctagagtc gacgggccgc ggtaacaatt gttaactaac ttaagctagc aacggtttcc
     3001 ctctagcggg atcaattccg cccccccccc ctaacgttac tggccgaagc cgcttggaat
     3061 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     3121 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     3181 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     3241 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     3301 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     3361 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     3421 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     3481 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     3541 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatgac cgagtacaag
     3601 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3661 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3721 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3781 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     3841 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     3901 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3961 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4021 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4081 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4141 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacccgg gcggggcgcg
     4201 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     4261 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     4321 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     4381 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     4441 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     4501 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4561 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     4621 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     4681 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     4741 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     4801 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     4861 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     4921 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     4981 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatat
     5041 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc
     5101 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
     5161 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag
     5221 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct
     5281 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg
     5341 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac
     5401 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt
     5461 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     5521 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     5581 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     5641 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     5701 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     5761 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     5821 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     5881 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     5941 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     6001 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     6061 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     6121 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga
     6181 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     6241 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     6301 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     6361 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     6421 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     6481 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     6541 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     6601 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     6661 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     6721 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     6781 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     6841 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     6901 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     6961 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     7021 cggtatcgcc tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     7081 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     7141 aaattttcgg gtttattaca gggacagcag agatccagtt tatcgatgag gccctttcgt
     7201 cttcactcga ggtgcccgtc agt
//
