LOCUS       sequence-405465-        6326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase reporter for Alu Domain Activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6326)
  TITLE     Borchert Lab Alu Domain Plasmids
REFERENCE   2  (bases 1 to 6326)
  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"
COMMENT     Sequence Label: psiCHECK-2 GATD1 Alu_WT
FEATURES             Location/Qualifiers
     source          1..6326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        62..253
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          489..621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     604..623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        666..684
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             694..1629
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1741..1789
                     /note="synthetic polyadenylation signal"
     promoter        1798..2549
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2579..2588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2585..4237
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             3544..3552
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4281..4402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4292..4311)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4346..4365
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4535..4639
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4640..4708,4709..5500)
                     /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(4858..4877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5671..6259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6160..6179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctgcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg ttaggtacct
       61 tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agcttgattc ttctgacaca acagtctcga acttaagctg cagaagttgg tcgtgaggca
      481 ctgggcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct
      541 tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc
      601 actttgcctt tctctccaca ggtgtccact cccagttcaa ttacagctct taaggctaga
      661 gtacttaata cgactcacta taggctagcc accatggctt ccaaggtgta cgaccccgag
      721 caacgcaaac gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg
      781 ctggactcct tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt
      841 ctgcatggta acgctgcctc cagctacctg tggaggcacg tcgtgcctca catcgagccc
      901 gtggctagat gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat
      961 ggctcatatc gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac
     1021 cttccaaaga aaatcatctt tgtgggccac gactgggggg cttgtctggc ctttcactac
     1081 tcctacgagc accaagacaa gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg
     1141 atcgagtcct gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa
     1201 gagggcgaga aaatggtgct tgagaataac ttcttcgtcg agaccatgct cccaagcaag
     1261 atcatgcgga aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag
     1321 ggcgaggtta gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc
     1381 aagcccgacg tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat
     1441 ctgcctaaga tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga
     1501 gctaagaagt tccctaacac cgagttcgtg aaggtgaagg gcctccactt cagccaggag
     1561 gacgctccag atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac
     1621 gagcagtaat tctaggcgat cgctcgagcc caaaccaaac cttagccggg cgcggtggtg
     1681 ggcgcctgta atcccattta cttgggaggc tgaggtttgg tttgggcggc cgctggccgc
     1741 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga ggatctaaat
     1801 gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct gacgcggggg
     1861 gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt cttggtggcc
     1921 acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc cgtggaggcg
     1981 ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc gcgcgcccca
     2041 gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg gcggcgacgg
     2101 gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg gagcatccgc
     2161 acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac gcggctgacg
     2221 cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat cttgtcaccc
     2281 ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat ccccgtggcc
     2341 cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt ctttagttct
     2401 atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac gcagatgcag
     2461 tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt ggcctcgaac
     2521 accgagcgac cctgcagcga cccgcttaaa agcttggcat tccggtactg ttggtaaagc
     2581 caccatggcc gatgctaaga acattaagaa gggccctgct cccttctacc ctctggagga
     2641 tggcaccgct ggcgagcagc tgcacaaggc catgaagagg tatgccctgg tgcctggcac
     2701 cattgccttc accgatgccc acattgaggt ggacatcacc tatgccgagt acttcgagat
     2761 gtctgtgcgc ctggccgagg ccatgaagag gtacggcctg aacaccaacc accgcatcgt
     2821 ggtgtgctct gagaactctc tgcagttctt catgccagtg ctgggcgccc tgttcatcgg
     2881 agtggccgtg gcccctgcta acgacattta caacgagcgc gagctgctga acagcatggg
     2941 catttctcag cctaccgtgg tgttcgtgtc taagaagggc ctgcagaaga tcctgaacgt
     3001 gcagaagaag ctgcctatca tccagaagat catcatcatg gactctaaga ccgactacca
     3061 gggcttccag agcatgtaca cattcgtgac atctcatctg cctcctggct tcaacgagta
     3121 cgacttcgtg ccagagtctt tcgacaggga caaaaccatt gccctgatca tgaacagctc
     3181 tgggtctacc ggcctgccta agggcgtggc cctgcctcat cgcaccgcct gtgtgcgctt
     3241 ctctcacgcc cgcgacccta ttttcggcaa ccagatcatc cccgacaccg ctattctgag
     3301 cgtggtgcca ttccaccacg gcttcggcat gttcaccacc ctgggctacc tgatttgcgg
     3361 ctttcgggtg gtgctgatgt accgcttcga ggaggagctg ttcctgcgca gcctgcaaga
     3421 ctacaaaatt cagtctgccc tgctggtgcc aaccctgttc agcttcttcg ctaagagcac
     3481 cctgatcgac aagtacgacc tgtctaacct gcacgagatt gcctctggcg gcgccccact
     3541 gtctaaggag gtgggcgaag ccgtggccaa gcgctttcat ctgccaggca tccgccaggg
     3601 ctacggcctg accgagacaa ccagcgccat tctgattacc ccagagggcg acgacaagcc
     3661 tggcgccgtg ggcaaggtgg tgccattctt cgaggccaag gtggtggacc tggacaccgg
     3721 caagaccctg ggagtgaacc agcgcggcga gctgtgtgtg cgcggcccta tgattatgtc
     3781 cggctacgtg aataaccctg aggccacaaa cgccctgatc gacaaggacg gctggctgca
     3841 ctctggcgac attgcctact gggacgagga cgagcacttc ttcatcgtgg accgcctgaa
     3901 gtctctgatc aagtacaagg gctaccaggt ggccccagcc gagctggagt ctatcctgct
     3961 gcagcaccct aacattttcg acgccggagt ggccggcctg cccgacgacg atgccggcga
     4021 gctgcctgcc gccgtcgtcg tgctggaaca cggcaagacc atgaccgaga aggagatcgt
     4081 ggactatgtg gccagccagg tgacaaccgc caagaagctg cgcggcggag tggtgttcgt
     4141 ggacgaggtg cccaagggcc tgaccggcaa gctggacgcc cgcaagatcc gcgagatcct
     4201 gatcaaggct aagaaaggcg gcaagatcgc cgtgtaataa ttctagagtc ggggcggccg
     4261 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4321 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4381 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4441 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     4501 taaggatcca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4561 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4621 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4681 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4741 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4801 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4861 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4921 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4981 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5041 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5101 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5161 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5221 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5281 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5341 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5401 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5461 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5521 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5581 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5641 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5701 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5761 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5821 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5881 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5941 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6001 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6061 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6121 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6181 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6241 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6301 ctggcctttt gctcacatgg ctcgac
//
