LOCUS       sequence-455836-        4772 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral vector for inducible expression of the green fluorescent 
            protein EGFP (TRE3G promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4772)
  TITLE     Morsut lab plasmids 2024
REFERENCE   2  (bases 1 to 4772)
  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"
COMMENT     Sequence Label: pHR_TRE3G_EGFP_WPRE
FEATURES             Location/Qualifiers
     source          1..4772
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             200..244
                     /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    529..646
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        708..1057
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    716..734
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    752..770
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    788..806
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    824..842
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    860..878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    896..914
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    932..950
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     996..1020
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(1063..1065,1066..1068,1069..1779)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1875..2463
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1928..1948)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2346..2357)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2466..2482)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2467..2483)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2573..2806
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(2831..2849)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2832..2849)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3139..3158)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3372..3391)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3491..3513
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3551..3569)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3637..3741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3742..3810,3811..4602)
                     /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(3960..3979)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
       61 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
      121 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
      181 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
      241 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
      301 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
      361 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
      421 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
      481 agtgaacgga tctcgacggt atcgccaaat ggcagtattc atccacaatt ttaaaagaaa
      541 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
      601 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
      661 ggacagcaga gatccagttt atcgatgagg ccctttcgtc ttcactcgag tttactccct
      721 atcagtgata gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag
      781 actttactcc ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga
      841 gaacgtatga ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct
      901 atcagtgata gagaacgtat atccagttta ctccctatca gtgatagaga acgtatgtcg
      961 aggtaggcgt gtacggtggg cgcctataaa agcagagctc gtttagtgaa ccgtcagatc
     1021 gcctggagca attccacaac acttttgtct tatacttacg cgatggtttc taagggtgaa
     1081 gaattgttca ccggagttgt gcccatcctg gttgagctgg acggagacgt taacggacac
     1141 aagttctcag tgagcggcga aggagagggc gacgctacgt acggtaaact gacgttgaag
     1201 ttcatctgta caacaggaaa gctgccagtg ccctggccta cactcgtgac caccctcacc
     1261 tacggagtcc agtgcttctc ccggtacccc gaccacatga aacagcacga tttcttcaag
     1321 tccgcgatgc ctgagggcta tgttcaggaa cgcaccatat tcttcaagga cgacggtaac
     1381 tataaaacca gggctgaagt taagttcgag ggggacacgt tggtaaatag aattgagttg
     1441 aaaggaatcg atttcaaaga agacggcaat attttgggcc acaagctgga gtacaactac
     1501 aacagccaca acgtgtatat tatggctgat aagcagaaga acggcatcaa ggtgaacttc
     1561 aaaatccgac acaatatcga ggatggctct gtgcaactgg ccgaccatta ccagcaaaac
     1621 acgcctatcg gcgacggtcc ggttttgctg ccagataacc actacctctc cacgcaatca
     1681 gccctctcca aggatcctaa cgaaaaaaga gaccatatgg ttctgcttga attcgtgacg
     1741 gctgctggga tcaccctggg aatggatgag ctgtataagg gccgcaactc ccacctgcaa
     1801 catgcgtgac tgactgaggc cgcgactcta gagtcgacct gcaggcatgc aagcttgata
     1861 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     1921 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     1981 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2041 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2101 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2161 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2221 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     2281 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2341 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2401 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2461 cgcatcgata ccgtcgacct cgagggaatt aattcgagct cggtaccttt aagaccaatg
     2521 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     2581 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     2641 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     2701 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     2761 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagcat ctagaattaa
     2821 ttccgtgtat tctatagtgt cacctaaatc gtatgtgtat gatacataag gttatgtatt
     2881 aattgtagcc gcgttctaac gacaatatgt acaagcctaa ttgtgtagca tctggcttac
     2941 tgaagcagac cctatcatct ctctcgtaaa ctgccgtcag agtcggtttg gttggacgaa
     3001 ccttctgagt ttctggtaac gccgtcccgc acccggaaat ggtcagcgaa ccaatcagca
     3061 gggtcatcgc tagccagatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     3121 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     3181 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     3241 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
     3301 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgaatgg
     3361 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     3421 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3481 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3541 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3601 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3661 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3721 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3781 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3841 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3901 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3961 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4021 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4081 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4141 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4201 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4261 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4321 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4381 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4441 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4501 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4561 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4621 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4681 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4741 tcagaccccg tagaaaagat caaaggatct tc
//
