LOCUS       sequence-446459-        8714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid expressing human VIM protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8714)
  TITLE     Lentiviral plasmids expressing popular genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8714)
  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            "Unpublished"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: pLenti puro HA-VIM
FEATURES             Location/Qualifiers
     source          1..8714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..69
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        51..69
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        97..323
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             324..504
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    551..676
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1169..1402
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1587..1631
                     /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"
     primer_bind     1929..1949
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        2238..2441
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2392..2412
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2489..2515
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3480..3491
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    4001..4118
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4167..4677
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     4596..4614
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             4699..5298
                     /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(4699..4718)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5195..5215
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5426..5659
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5731..5852
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5768..5787)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5822..5841
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5892..6027
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6047..6066)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6048..6066)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6098..6115)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6098..6114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6107..6129)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6323..6342)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6442..6464
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6502..6520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6588..6692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6693..6761,6762..7553)
                     /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     complement(6911..6930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7724..8312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8213..8232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8466..8483
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8600..8621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8636..8641,8642..8659,8660..8666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8674..8690
                     /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     8679..8701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8698..8714
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8698..8714
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgattacg ccaagcttgc atgcctgcag catgattacg ccaagcgcgc aattaaccct
       61 cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt
      121 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
      181 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
      241 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
      301 agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc
      361 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
      421 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
      481 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
      541 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
      601 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
      661 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
      721 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
      781 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
      841 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
      901 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
      961 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
     1021 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     1081 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1141 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1201 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1261 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1321 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1381 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     1441 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     1501 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     1561 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     1621 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     1681 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     1741 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     1801 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     1861 ttcgattagt gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc
     1921 ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     1981 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     2041 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     2101 actatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2161 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2221 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2281 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2341 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2401 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     2461 tccgctagcg ctaccggtcg ccaccatgta cccatacgat gttccggatt acgctagcgg
     2521 atccatgtct acaaggagtg tgagtagttc ttcttaccga cgaatgtttg gcggtcctgg
     2581 tacggcgagc aggcccagta gctctcgaag ctacgtgact acgtccacgc gcacttacag
     2641 tctcgggagt gcactgagac cctcaaccag tcgctctctg tatgctagtt cccccggcgg
     2701 agtatatgcg acgaggtcct cagccgttcg gctccggtca tccgttcccg gcgttaggct
     2761 gttgcaggac agcgttgatt tttcactcgc tgatgcaatt aataccgaat ttaagaacac
     2821 taggacgaac gaaaaagtgg aattgcagga gcttaatgat cgatttgcta actatatcga
     2881 taaggtgcga ttccttgaac agcaaaacaa aattcttctg gcagaacttg agcagctcaa
     2941 ggggcaggga aagtcacgcc ttggagactt gtacgaagaa gaaatgaggg aactgcgccg
     3001 acaagtagac cagcttacga atgacaaggc gagagtagag gtagagaggg ataatctcgc
     3061 cgaagatata atgcgcctcc gagaaaagtt gcaagaggaa atgttgcaac gcgaggaggc
     3121 agagaataca ctccaatcct ttagacagga tgttgacaat gcaagcctgg cccgcttgga
     3181 ccttgaacgc aaggtagaat cccttcagga ggaaattgct tttctgaaga agctccacga
     3241 agaagaaatc caagagttgc aagcccaaat ccaggaacag catgtacaga tagacgtcga
     3301 tgtatcaaaa cccgacctga cggcggcatt gcgagatgtt agacaacagt atgagtccgt
     3361 cgctgccaag aatcttcaag aggcagaaga atggtacaag tctaaatttg ccgacctgag
     3421 tgaagcagca aataggaata acgacgccct tcggcaagcg aaacaggaat ctacggagta
     3481 tcgaaggcag gttcagtcat tgacctgtga agtagatgcc ctcaaaggta ctaacgaaag
     3541 cctggaacgg cagatgcgcg agatggagga aaatttcgcc gtggaagctg ctaattatca
     3601 agacacaatt ggacgccttc aagacgagat ccagaacatg aaagaagaaa tggccaggca
     3661 cctcagggaa tatcaggatc tccttaacgt taagatggcg cttgatatag agatcgctac
     3721 ataccgaaag ctgttggaag gggaggagag taggatttca ctgcctcttc caaatttcag
     3781 ttctttgaac ttgcgagaaa ccaaccttga ttctttgcca cttgtcgaca cccactccaa
     3841 aagaacactc ctcatcaaga cggttgaaac cagggatggc caggtgatta atgaaacatc
     3901 acagcaccac gatgacttgg aatagccgga ctcagatctc gagctcaagc ttctctagag
     3961 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     4021 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     4081 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     4141 gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca aggcagccct
     4201 gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag cggcgccgac
     4261 cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt cgccgctacc
     4321 cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg ttccttgcgg
     4381 ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac cctcgcagac
     4441 ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc caatagcggc
     4501 tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga ggcggggtgt
     4561 ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg caagcctccg
     4621 gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc tccccagggg
     4681 gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4741 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4801 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4861 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4921 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4981 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     5041 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     5101 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     5161 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     5221 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     5281 cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga aaggagcgca
     5341 cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     5401 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     5461 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5521 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5581 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5641 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     5701 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     5761 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5821 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     5881 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5941 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6001 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     6061 gtattacgcg cgctccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc
     6121 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6181 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6241 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     6301 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     6361 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     6421 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     6481 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     6541 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     6601 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     6661 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6721 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6781 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6841 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6901 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6961 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7021 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7081 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7141 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7201 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7261 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7321 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7381 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7441 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     7501 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     7561 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     7621 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     7681 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7741 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7801 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7861 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7921 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7981 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8041 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8101 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8161 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8221 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8281 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8341 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8401 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     8461 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     8521 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     8581 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     8641 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     8701 gaaacagcta tgac
//
