LOCUS       sequence-420415-        9131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Positive control vector for luciferase lentivirus-based enhancer 
            assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9131)
  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 9131)
  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-luc
FEATURES             Location/Qualifiers
     source          1..9131
                     /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      3453..3462
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3459..5111
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     protein_bind    complement(5131..5164)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5220..5808
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5273..5293)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5691..5702)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5811..5827)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5812..5828)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7137..7317
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7379..7967)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7459..7478)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8138..8929,8930..8998))
                     /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     8761..8780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8999..9103)
                     /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 gactatttac 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 actgttggta aagccaccat ggaagatgcc aaaaacatta
     3481 agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag cagctgcaca
     3541 aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac gcacatatcg
     3601 aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca gaagctatga
     3661 agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat agcttgcagt
     3721 tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca gctaacgaca
     3781 tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc gtcgtattcg
     3841 tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg atcatacaaa
     3901 agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg tacaccttcg
     3961 tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag agcttcgacc
     4021 gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg cccaagggcg
     4081 tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac cccatcttcg
     4141 gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac cacggcttcg
     4201 gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc atgtaccgct
     4261 tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct gccctgctgg
     4321 tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac gacctaagca
     4381 acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt gaggccgtgg
     4441 ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa acaaccagcg
     4501 ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag gtggtgccct
     4561 tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg aaccagcgcg
     4621 gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac cccgaggcta
     4681 caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc tactgggacg
     4741 aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac aagggctacc
     4801 aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc ttcgacgccg
     4861 gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc gtcgtgctgg
     4921 aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc caggttacaa
     4981 ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa ggactgaccg
     5041 gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag ggcggcaaga
     5101 tcgccgtgta agaattcgtc gagggaccta ataacttcgt atagcataca ttatacgaag
     5161 ttatacatgt ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata
     5221 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5281 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5341 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5401 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5461 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     5521 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5581 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5641 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5701 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5761 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     5821 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     5881 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     5941 cctcaggtac caagcatggg gtaaagtact gttctcatca catcatatca aggttatata
     6001 ccatcaatat tgccacagat gttacttagc cttttaatat ttctctaatt tagtgtatat
     6061 gcaatgatag ttctctgatt tctgagattg agtttctcat gtgtaatgat tatttagagt
     6121 ttctctttca tctgttcaaa tttttgtcta gttttatttt ttactgattt gtaagacttc
     6181 tttttataat ctgcatatta caattctctt tactggggtg ttgcaaatat tttctgtcat
     6241 tctatggcct gacttttctt aatggttttt taattttaaa aataagtctt aatattcatg
     6301 caatctaatt aacaatcttt tctttgtggt taggactttg agtcataaga aatttttctc
     6361 tacactgaag tcatgatggc atgcttctat attattttct aaaagattta aagttttgcc
     6421 ttctccattt agacttataa ttcactggaa tttttttgtg tgtatggtat gacatatggg
     6481 ttccctttta ttttttacat ataaatatat ttccctgttt ttctaaaaaa gaaaaagatc
     6541 atcattttcc cattgtaaaa tgccatattt ttttcatagg tcacttacat atatcaatgg
     6601 gtctgtttct gagctctact ctattttatc agcctcactg tctatcccca cacatctcat
     6661 gctttgctct aaatcttgat atttagtgga acattctttc ccattttgtt ctacaagaat
     6721 atttttgtta ttgtcttttg ggcttctata tacattttag aatgaggttg gcaagttctg
     6781 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6841 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
     6901 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
     6961 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
     7021 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
     7081 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
     7141 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     7201 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     7261 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca
     7321 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7381 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7441 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7501 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7561 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7621 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7681 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7741 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7801 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7861 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7921 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7981 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     8041 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     8101 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     8161 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     8221 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     8281 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     8341 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     8401 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     8461 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     8521 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     8581 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8641 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8701 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8761 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8821 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8881 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8941 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     9001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     9061 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     9121 tgccacctga c
//
