LOCUS       sequence-123983-        8507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase in lentivirus backbone to report Dux4 activation of 
            RFPL4b promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8507)
  TITLE     Dux4 reporters
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8507)
  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            "Unpublished"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.cPPT.RFPL4b.Luciferase.WPRE
FEATURES             Location/Qualifiers
     source          1..8507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      734..743
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             740..2392
                     /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(806..824)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     misc_feature    2404..2992
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2457..2477)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2875..2886)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3079..3312
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3384..3505
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3421..3440)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3475..3494
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3524..3659
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3691..3710)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3692..3710)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3720..3737)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3720..3736)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3729..3751)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3881..4336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3968..3987)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4178..4199
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4365..4469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4470..4538,4539..5330)
                     /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(4688..4707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5501..6089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5990..6009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6243..6260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6377..6398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6413..6418,6419..6436,6437..6443)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6451..6467
                     /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     6456..6478
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6475..6491
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6475..6491
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6510..6530
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6512..6530
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             6786..6966
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7010..7135
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7628..7861
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8046..8090
                     /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    8357..8474
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctctgttgtg acttttactg ctccttctgc tctcgataca gacaccataa aatttaccgt
       61 tcatccaaga acacttttga gagtaaaaaa tggaattatt aataattaca ggacaatagc
      121 cttgcctagc tgccatcttt cttatcctgt ccagtaagta cccctaagag tagaattgtt
      181 gagtcaaagg acatgaaaac tgttgtagat cctgtccgct catgattcta tcaggaatag
      241 gaacaccaat gattacatta gccacgaggt gattaggttt cagcagtgca tttctgtatt
      301 tgatctccaa cacattttgc tttttaaagt tgagttctga aattagagac aagagcagat
      361 ggcagtagat tcactctaat tcagggagtt attaagaaat tttgaatttt tttactatat
      421 aatttgaagc ctgctttaat gtcatcatca taaacggaag ggtgggttga tgagtttact
      481 cagattagag tggtacaacc tttggggcgg agcctctgac tctgattggt tggacaatgt
      541 tttccaggca gtataaaaac gctggtgtct ctccgcggaa actccgagag gagcagaggt
      601 ctgtagaggt agagacgtag gcttcggatc ttttagaatt ctgctggaag tctccaagga
      661 gctcttacgc gtgctagccc gggctcgaga tctgcgatct aagtaagctt ggcattccgg
      721 tactgttggt aaagccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt
      781 ctatccgctg gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc
      841 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc
      901 tgagtacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac
      961 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg
     1021 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt
     1081 gctcaacagt atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca
     1141 aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc
     1201 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc
     1261 cggttttaat gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact
     1321 gatcatgaac tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac
     1381 tgcctgcgtg agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga
     1441 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg
     1501 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct
     1561 gaggagcctt caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt
     1621 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc
     1681 tggtggcgct cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc
     1741 aggtatcagg caaggatatg ggctcactga gactacatca gctattctga ttacacccga
     1801 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt
     1861 ggatctggat accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg
     1921 tcctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa
     1981 ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat
     2041 cgttgaccgc ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt
     2101 ggaatccatc ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga
     2161 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac
     2221 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg
     2281 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa
     2341 aatcagagag atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagtc
     2401 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2461 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2521 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2581 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2641 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2701 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2761 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     2821 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2881 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     2941 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     3001 cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     3061 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     3121 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     3181 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     3241 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     3301 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     3361 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     3421 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3481 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     3541 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3601 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3661 aggcttttgc gtcgagacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     3721 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3781 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3841 ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca
     3901 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3961 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4021 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4081 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4141 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4201 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     4261 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4321 ttaacgttta caatttccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4441 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4501 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4561 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4621 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4681 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4741 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4801 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4861 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4921 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4981 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5641 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5881 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6241 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     6301 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     6361 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     6421 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     6481 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     6541 tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac
     6601 gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga
     6661 agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg
     6721 acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa
     6781 taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6841 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6901 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6961 ctagcagtgg cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc tctctcgacg
     7021 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     7081 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     7141 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     7201 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     7261 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     7321 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     7381 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     7441 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     7501 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     7561 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     7621 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     7681 cctcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     7741 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     7801 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     7861 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     7921 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     7981 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     8041 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     8101 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     8161 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     8221 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     8281 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatctc
     8341 gacggtatcg gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     8401 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     8461 aaattcaaaa ttttatcgat cacgagacta gcctcgagaa gcttgat
//
