LOCUS       sequence-455835-        4908 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 4908)
  TITLE     Morsut lab plasmids 2024
REFERENCE   2  (bases 1 to 4908)
  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..4908
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             65..109
                     /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    394..511
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        573..922
                     /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    581..599
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    617..635
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    653..671
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    689..707
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    725..743
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    761..779
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    797..815
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     861..885
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      963..972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1032..1034,1035..1037,1038..1748)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2011..2599
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2064..2084)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2482..2493)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2602..2618)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2603..2619)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2709..2942
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(2967..2985)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2968..2985)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3275..3294)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3508..3527)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3627..3649
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3687..3705)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3773..3877
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3878..3946,3947..4738)
                     /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(4096..4115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
       61 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
      121 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
      181 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
      241 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
      301 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
      361 acggtatcgc caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
      421 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
      481 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
      541 agtttatcga tgaggccctt tcgtcttcac tcgagtttac tccctatcag tgatagagaa
      601 cgtatgaaga gtttactccc tatcagtgat agagaacgta tgcagacttt actccctatc
      661 agtgatagag aacgtataag gagtttactc cctatcagtg atagagaacg tatgaccagt
      721 ttactcccta tcagtgatag agaacgtatc tacagtttac tccctatcag tgatagagaa
      781 cgtatatcca gtttactccc tatcagtgat agagaacgta tgtcgaggta ggcgtgtacg
      841 gtgggcgcct ataaaagcag agctcgttta gtgaaccgtc agatcgcctg gagcaattcc
      901 acaacacttt tgtcttatac ttacgcgtca agtggagcaa ggcaggtgga cagtggatcg
      961 ccgccaccat ggagaccgac accctcttgc tctgggtcct gctgctgtgg gttccgggta
     1021 gtaccggaga catggtttct aagggtgaag aattgttcac cggagttgtg cccatcctgg
     1081 ttgagctgga cggagacgtt aacggacaca agttctcagt gagcggcgaa ggagagggcg
     1141 acgctacgta cggtaaactg acgttgaagt tcatctgtac aacaggaaag ctgccagtgc
     1201 cctggcctac actcgtgacc accctcacct acggagtcca gtgcttctcc cggtaccccg
     1261 accacatgaa acagcacgat ttcttcaagt ccgcgatgcc tgagggctat gttcaggaac
     1321 gcaccatatt cttcaaggac gacggtaact ataaaaccag ggctgaagtt aagttcgagg
     1381 gggacacgtt ggtaaataga attgagttga aaggaatcga tttcaaagaa gacggcaata
     1441 ttttgggcca caagctggag tacaactaca acagccacaa cgtgtatatt atggctgata
     1501 agcagaagaa cggcatcaag gtgaacttca aaatccgaca caatatcgag gatggctctg
     1561 tgcaactggc cgaccattac cagcaaaaca cgcctatcgg cgacggtccg gttttgctgc
     1621 cagataacca ctacctctcc acgcaatcag ccctctccaa ggatcctaac gaaaaaagag
     1681 accatatggt tctgcttgaa ttcgtgacgg ctgctgggat caccctggga atggatgagc
     1741 tgtataagaa cgccgtgggc caggatacgc aggaggtgat cgtcgtgccc cacagcttgc
     1801 ccttcaaggt cgtggttatt agtgctatcc tggcgttggt ggtgctgact attatctccc
     1861 ttatcatact tatcatgctg tggcaaaaaa aaccccgata agatccttga cttgcggccg
     1921 caactcccac ctgcaacatg cgtgactgac tgaggccgcg actctagagt cgacctgcag
     1981 gcatgcaagc ttgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     2041 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2101 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2161 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2221 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2281 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2341 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2401 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2461 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2521 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2581 ctttgggccg cctccccgca tcgataccgt cgacctcgag ggaattaatt cgagctcggt
     2641 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     2701 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     2761 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     2821 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     2881 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     2941 cagcatctag aattaattcc gtgtattcta tagtgtcacc taaatcgtat gtgtatgata
     3001 cataaggtta tgtattaatt gtagccgcgt tctaacgaca atatgtacaa gcctaattgt
     3061 gtagcatctg gcttactgaa gcagacccta tcatctctct cgtaaactgc cgtcagagtc
     3121 ggtttggttg gacgaacctt ctgagtttct ggtaacgccg tcccgcaccc ggaaatggtc
     3181 agcgaaccaa tcagcagggt catcgctagc cagatcctct acgccggacg catcgtggcc
     3241 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     3301 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     3361 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     3421 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     3481 ggagagcgtc gaatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3541 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3601 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3661 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3721 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3781 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3841 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3901 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3961 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4021 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4081 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4141 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4201 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4261 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4321 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4381 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4441 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4501 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4561 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4621 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4681 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4741 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4801 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4861 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
