LOCUS       sequence-421904-        8447 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-driven Renilla luciferase expression vector for luciferase 
            lentivirus-based reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8447)
  AUTHORS   Deng C, Whalen S, Steyert M, Ziffra R, Przytycki PF, Inoue F, 
            Pereira DA, Capauto D, Norton S, Vaccarino FM, Pollen AA, Nowakowski
            TJ, Ahituv N, Pollard KS
  TITLE     Massively parallel characterization of regulatory elements in the 
            developing human cortex.
  JOURNAL   Science. 2024 May 24;384(6698):eadh0559. doi: 
            10.1126/science.adh0559. Epub 2024 May 24.
  PUBMED    38781390
REFERENCE   2  (bases 1 to 8447)
  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: "Science."; 
            date: "2024-05-24"; volume: "384(6698):eadh0559. doi"; pages: " 
            10.1126/science.adh0559. Epub 2024 May 24"
COMMENT     Sequence Label: pLS-CMV-mP-Rluc
FEATURES             Location/Qualifiers
     source          1..8447
                     /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"
     enhancer        3075..3378
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3395..3426
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      3466..3475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3472..4407
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     protein_bind    complement(4447..4480)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4536..5124
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4589..4609)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5007..5018)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5127..5143)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5128..5144)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6453..6633
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6695..7283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6775..6794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7454..8245,8246..8314))
                     /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     8077..8096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8315..8419)
                     /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 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     3061 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     3121 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     3181 ttccattgac gtcaatgggt ggactattta cggtaaactg cccacttggc agtacatcaa
     3241 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     3301 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     3361 gtcatcgcta ttaccatgct agacctgcag gcactagagg gtatataatg gaagctcgac
     3421 ttccagcttg gcaatccggt actgtaccgg tgtactgttg gtaaagccac catggcttcc
     3481 aaggtgtacg accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc
     3541 tgcaagcaaa tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc
     3601 gagaacgccg tgatttttct gcatggtaac gctgcctcca gctacctgtg gaggcacgtc
     3661 gtgcctcaca tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag
     3721 tccggcaaga gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct
     3781 tggttcgagc tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct
     3841 tgtctggcct ttcactactc ctacgagcac caagacaaga tcaaggccat cgtccatgct
     3901 gagagtgtcg tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc
     3961 gccctgatca agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag
     4021 accatgctcc caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg
     4081 gagccattca aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc
     4141 cctctcgtta agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac
     4201 cttcgggcca gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg gttcttttcc
     4261 aacgctattg tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc
     4321 ctccacttca gccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg
     4381 gagcgcgtgc tgaagaacga gcagtaattc tagagtcggg gcggccggaa ttcgtcgagg
     4441 gacctaataa cttcgtatag catacattat acgaagttat acatgtttaa gggttccggt
     4501 tccactaggt acaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     4561 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     4621 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     4681 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     4741 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     4801 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     4861 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     4921 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     4981 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     5041 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     5101 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa
     5161 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
     5221 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccaag catggggtaa
     5281 agtactgttc tcatcacatc atatcaaggt tatataccat caatattgcc acagatgtta
     5341 cttagccttt taatatttct ctaatttagt gtatatgcaa tgatagttct ctgatttctg
     5401 agattgagtt tctcatgtgt aatgattatt tagagtttct ctttcatctg ttcaaatttt
     5461 tgtctagttt tattttttac tgatttgtaa gacttctttt tataatctgc atattacaat
     5521 tctctttact ggggtgttgc aaatattttc tgtcattcta tggcctgact tttcttaatg
     5581 gttttttaat tttaaaaata agtcttaata ttcatgcaat ctaattaaca atcttttctt
     5641 tgtggttagg actttgagtc ataagaaatt tttctctaca ctgaagtcat gatggcatgc
     5701 ttctatatta ttttctaaaa gatttaaagt tttgccttct ccatttagac ttataattca
     5761 ctggaatttt tttgtgtgta tggtatgaca tatgggttcc cttttatttt ttacatataa
     5821 atatatttcc ctgtttttct aaaaaagaaa aagatcatca ttttcccatt gtaaaatgcc
     5881 atattttttt cataggtcac ttacatatat caatgggtct gtttctgagc tctactctat
     5941 tttatcagcc tcactgtcta tccccacaca tctcatgctt tgctctaaat cttgatattt
     6001 agtggaacat tctttcccat tttgttctac aagaatattt ttgttattgt cttttgggct
     6061 tctatataca ttttagaatg aggttggcaa gttctgtaga tcttagccac tttttaaaag
     6121 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
     6181 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
     6241 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
     6301 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
     6361 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
     6421 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
     6481 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6541 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6601 gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc cagcaaaagg
     6661 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6721 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6781 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6841 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6901 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6961 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     7021 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     7081 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     7141 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7201 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7261 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7321 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7381 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7441 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     7501 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7561 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7621 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7681 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7741 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7801 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7861 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7921 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7981 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     8041 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     8101 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     8161 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     8221 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     8281 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     8341 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     8401 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
