LOCUS       sequence-449975-        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  JOURNAL   Unpublished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            "Unpublished"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     332..352
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             563..796
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    868..989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(905..924)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     959..978
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1029..1164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1184..1203)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1185..1203)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1235..1252)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1235..1251)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1244..1266)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1460..1479)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1579..1601
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1639..1657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1725..1829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1830..1898,1899..2690)
                     /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(2048..2067)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2861..3449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3350..3369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3603..3620
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3737..3758
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3773..3778,3779..3796,3797..3803)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3811..3827
                     /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     3816..3838
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3835..3851
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3835..3851
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3900..3920
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3902..3920
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3948..4174
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4175..4355
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4402..4527
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5020..5253
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5438..5482
                     /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     5780..5800
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        6089..6292
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6243..6263
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             6340..6366
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7331..7342
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    7852..7969
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8018..8528
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     8447..8465
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(8550..8714,1..435)
                     /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(8550..8569)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
       61 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
      121 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
      181 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
      241 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
      301 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
      361 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
      421 aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga
      481 ccccatgcat cggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
      541 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
      601 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      661 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      721 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      781 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
      841 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
      901 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
      961 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     1021 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     1081 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     1141 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     1201 ttacgcgcgc tccccgggta ccgagctcga attcactggc cgtcgtttta caacgtcgtg
     1261 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1321 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1381 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     1441 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     1501 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     1561 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     1621 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1681 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1741 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1801 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1861 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     1921 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1981 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2041 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2101 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2161 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2221 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2281 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2341 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2401 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2461 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2521 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2581 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2641 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2701 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     2761 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     2821 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2881 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2941 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3001 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3061 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3121 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3181 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3241 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3301 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3361 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3421 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3481 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3541 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3601 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3661 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3721 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     3781 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     3841 acagctatga ccatgattac gccaagcttg catgcctgca gcatgattac gccaagcgcg
     3901 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
     3961 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
     4021 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
     4081 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
     4141 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
     4201 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     4261 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     4321 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
     4381 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     4441 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     4501 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     4561 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     4621 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     4681 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     4741 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     4801 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     4861 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     4921 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     4981 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     5041 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     5101 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     5161 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     5221 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     5281 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     5341 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     5401 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     5461 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     5521 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     5581 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     5641 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     5701 agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg
     5761 agcggccgcc cccttcaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg
     5821 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
     5881 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     5941 ttttaaaatg gactatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     6001 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     6061 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     6121 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     6181 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     6241 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     6301 gaaccgtcag atccgctagc gctaccggtc gccaccatgt acccatacga tgttccggat
     6361 tacgctagcg gatccatgtc tacaaggagt gtgagtagtt cttcttaccg acgaatgttt
     6421 ggcggtcctg gtacggcgag caggcccagt agctctcgaa gctacgtgac tacgtccacg
     6481 cgcacttaca gtctcgggag tgcactgaga ccctcaacca gtcgctctct gtatgctagt
     6541 tcccccggcg gagtatatgc gacgaggtcc tcagccgttc ggctccggtc atccgttccc
     6601 ggcgttaggc tgttgcagga cagcgttgat ttttcactcg ctgatgcaat taataccgaa
     6661 tttaagaaca ctaggacgaa cgaaaaagtg gaattgcagg agcttaatga tcgatttgct
     6721 aactatatcg ataaggtgcg attccttgaa cagcaaaaca aaattcttct ggcagaactt
     6781 gagcagctca aggggcaggg aaagtcacgc cttggagact tgtacgaaga agaaatgagg
     6841 gaactgcgcc gacaagtaga ccagcttacg aatgacaagg cgagagtaga ggtagagagg
     6901 gataatctcg ccgaagatat aatgcgcctc cgagaaaagt tgcaagagga aatgttgcaa
     6961 cgcgaggagg cagagaatac actccaatcc tttagacagg atgttgacaa tgcaagcctg
     7021 gcccgcttgg accttgaacg caaggtagaa tcccttcagg aggaaattgc ttttctgaag
     7081 aagctccacg aagaagaaat ccaagagttg caagcccaaa tccaggaaca gcatgtacag
     7141 atagacgtcg atgtatcaaa acccgacctg acggcggcat tgcgagatgt tagacaacag
     7201 tatgagtccg tcgctgccaa gaatcttcaa gaggcagaag aatggtacaa gtctaaattt
     7261 gccgacctga gtgaagcagc aaataggaat aacgacgccc ttcggcaagc gaaacaggaa
     7321 tctacggagt atcgaaggca ggttcagtca ttgacctgtg aagtagatgc cctcaaaggt
     7381 actaacgaaa gcctggaacg gcagatgcgc gagatggagg aaaatttcgc cgtggaagct
     7441 gctaattatc aagacacaat tggacgcctt caagacgaga tccagaacat gaaagaagaa
     7501 atggccaggc acctcaggga atatcaggat ctccttaacg ttaagatggc gcttgatata
     7561 gagatcgcta cataccgaaa gctgttggaa ggggaggaga gtaggatttc actgcctctt
     7621 ccaaatttca gttctttgaa cttgcgagaa accaaccttg attctttgcc acttgtcgac
     7681 acccactcca aaagaacact cctcatcaag acggttgaaa ccagggatgg ccaggtgatt
     7741 aatgaaacat cacagcacca cgatgacttg gaatagccgg actcagatct cgagctcaag
     7801 cttctctaga gaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
     7861 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     7921 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     7981 cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc
     8041 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
     8101 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
     8161 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
     8221 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
     8281 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
     8341 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
     8401 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
     8461 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     8521 ctccccaggg ggatccaccg gagcttacca tgaccgagta caagcccacg gtgcgcctcg
     8581 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     8641 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     8701 aactcttcct cacg
//
