LOCUS       sequence-122039-        7809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luciferase reporter in lentivirus backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7809)
  TITLE     Dux4 reporters
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7809)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pRRLSIN.cPPTLuciferase.WPRE
FEATURES             Location/Qualifiers
     source          1..7809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      50..59
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             56..1708
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(122..140)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    1720..2308
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1773..1793)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2191..2202)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2395..2628
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2700..2821
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2737..2756)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2791..2810
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2840..2975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3007..3026)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3008..3026)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3036..3053)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3036..3052)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3045..3067)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3197..3652
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3284..3303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3494..3515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3681..3785
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3786..3854,3855..4646)
                     /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(4004..4023)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4817..5405
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5306..5325
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5559..5576
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5693..5714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5729..5734,5735..5752,5753..5759)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5767..5783
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5772..5794
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5791..5807
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5791..5807
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5826..5846
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5828..5846
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             6102..6282
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6326..6451
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6944..7177
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7362..7406
                     /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    7673..7790
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tcgagatctg cgatctaagt aagcttggca ttccggtact gttggtaaag ccaccatgga
       61 agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc
      121 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
      181 tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg
      241 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
      301 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
      361 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca
      421 gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
      481 gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca
      541 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
      601 gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac
      661 tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc
      721 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
      781 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
      841 cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat
      901 tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga
      961 caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga
     1021 agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct
     1081 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     1141 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     1201 gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt
     1261 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     1321 catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat
     1381 taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc
     1441 caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc
     1501 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     1561 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     1621 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     1681 caagaagggc ggaaagatcg ccgtgtaatt ctagtcgaca atcaacctct ggattacaaa
     1741 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1801 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     1861 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     1921 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     1981 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2041 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2101 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     2161 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2221 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2281 cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacct ttaagaccaa
     2341 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     2401 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     2641 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     2701 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2761 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2821 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     2881 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     2941 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcgtcg agacgtaccc
     3001 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     3061 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     3121 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3181 aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3241 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3301 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3361 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3421 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3481 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3541 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3601 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcccaggtg
     3661 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3721 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3781 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3841 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3901 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3961 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4021 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4081 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4141 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4201 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4261 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4321 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4381 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4441 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4501 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4561 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4621 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4681 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4741 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4801 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4861 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4921 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4981 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5041 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5101 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5161 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5221 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5281 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5341 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5401 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5461 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5521 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5581 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5641 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5701 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5761 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     5821 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
     5881 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
     5941 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
     6001 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
     6061 cagagatatt gtatttaagt gcctagctcg atacaataaa cgggtctctc tggttagacc
     6121 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     6181 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     6241 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     6301 cctgaaagcg aaagggaaac cagagctctc tcgacgcagg actcggcttg ctgaagcgcg
     6361 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     6421 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     6481 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     6541 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     6601 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     6661 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     6721 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     6781 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     6841 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     6901 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     6961 ggttcttggg agcagcagga agcactatgg gcgcagcctc aatgacgctg acggtacagg
     7021 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     7081 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     7141 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     7201 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     7261 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     7321 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     7381 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     7441 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     7501 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     7561 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     7621 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcggtta acttttaaaa
     7681 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     7741 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt atcgatcacg
     7801 agactagcc
//
