LOCUS       sequence-315921-        7816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus enhancer assay plasmid that contains a minimal promoter 
            (mP) and a EGFP reporter gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7816)
  AUTHORS   Inoue F, Kircher M, Martin B, Cooper GM, Witten DM, McManus MT, 
            Ahituv N, Shendure J
  TITLE     A systematic comparison reveals substantial differences in 
            chromosomal versus episomal encoding of enhancer activity.
  JOURNAL   Genome Res. 2016 Nov 9. pii: gr.212092.116.
  PUBMED    27831498
REFERENCE   2  (bases 1 to 7816)
  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: "Genome Res.
            2016 Nov 9. pii: gr.212092.116."
COMMENT     Sequence Label: pLS-mP
FEATURES             Location/Qualifiers
     source          1..7816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3013..3044
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      3071..3080
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3077..3079,3080..3082,3083..3796)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3077..3079,3080..3082,3083..3793)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3122..3143)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3383..3402)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3730..3751
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3816..3849)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3905..4493
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3958..3978)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4376..4387)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4496..4512)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4497..4513)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5822..6002
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6064..6652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6144..6163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6823..7614,7615..7683))
                     /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     7446..7465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7684..7788)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cggtgctttg ctctgagcca gcccaccagt
     2641 ttggaatgac tcctttttat gacttgaatt ttcaagtata aagtctagtg ctaaatttaa
     2701 tttgaacaac tgtatagttt ttgctggttg ggggaaggaa aaaaaatggt ggcagtgttt
     2761 ttttcagaat tagaagtgaa atgaaaactt gttgtgtgtg aggatttcta atgacatgtg
     2821 gtggttgcat actgagtgaa gccggtgagc attctgccat gtcaccccct cgtgctcagt
     2881 aatgtacttt acagaaatcc taaactcaaa agattgatat aaaccatgct tcttgtgtat
     2941 atccggtctc ttctctgggt agtctcactc agcctgcatt tctgccaggg cccgctctag
     3001 acctgcaggc actagagggt atataatgga agctcgactt ccagcttggc aatccggtac
     3061 tgtaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3121 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3181 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3241 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3301 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3361 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3421 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3481 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3541 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3601 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3661 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3721 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3781 cgagctgtac aagtaggaat tcgtcgaggg acctaataac ttcgtatagc atacattata
     3841 cgaagttata catgtttaag ggttccggtt ccactaggta caattcgata tcaagcttat
     3901 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3961 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     4021 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     4081 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     4141 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     4201 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4261 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     4321 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     4381 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     4441 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     4501 ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     4561 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     4621 tcacacctca ggtaccaagc atggggtaaa gtactgttct catcacatca tatcaaggtt
     4681 atataccatc aatattgcca cagatgttac ttagcctttt aatatttctc taatttagtg
     4741 tatatgcaat gatagttctc tgatttctga gattgagttt ctcatgtgta atgattattt
     4801 agagtttctc tttcatctgt tcaaattttt gtctagtttt attttttact gatttgtaag
     4861 acttcttttt ataatctgca tattacaatt ctctttactg gggtgttgca aatattttct
     4921 gtcattctat ggcctgactt ttcttaatgg ttttttaatt ttaaaaataa gtcttaatat
     4981 tcatgcaatc taattaacaa tcttttcttt gtggttagga ctttgagtca taagaaattt
     5041 ttctctacac tgaagtcatg atggcatgct tctatattat tttctaaaag atttaaagtt
     5101 ttgccttctc catttagact tataattcac tggaattttt ttgtgtgtat ggtatgacat
     5161 atgggttccc ttttattttt tacatataaa tatatttccc tgtttttcta aaaaagaaaa
     5221 agatcatcat tttcccattg taaaatgcca tatttttttc ataggtcact tacatatatc
     5281 aatgggtctg tttctgagct ctactctatt ttatcagcct cactgtctat ccccacacat
     5341 ctcatgcttt gctctaaatc ttgatattta gtggaacatt ctttcccatt ttgttctaca
     5401 agaatatttt tgttattgtc ttttgggctt ctatatacat tttagaatga ggttggcaag
     5461 ttctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     5521 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     5581 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     5641 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     5701 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     5761 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     5821 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     5881 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     5941 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6001 cagcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6061 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6241 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6481 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7741 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7801 aaaagtgcca cctgac
//
