LOCUS       sequence-405408-        6327 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Luciferase reporter for Alu Domain Activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6327)
  TITLE     Borchert Lab Alu Domain Plasmids
REFERENCE   2  (bases 1 to 6327)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_Mut2
FEATURES             Location/Qualifiers
     source          1..6327
                     /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    1742..1790
                     /note="synthetic polyadenylation signal"
     promoter        1799..2550
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      2580..2589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2586..4238
                     /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             3545..3553
                     /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    4282..4403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4293..4312)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4347..4366
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4536..4640
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4641..4709,4710..5501)
                     /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(4859..4878)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5672..6260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6161..6180
                     /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 cgcacacaca
     1681 cgcgcctgta atcccaacac actgggaggc tgaggtttgg tttggggcgg ccgctggccg
     1741 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aggatctaaa
     1801 tgagtcttcg gacctcgcgg gggccgctta agcggtggtt agggtttgtc tgacgcgggg
     1861 ggagggggaa ggaacgaaac actctcattc ggaggcggct cggggtttgg tcttggtggc
     1921 cacgggcacg cagaagagcg ccgcgatcct cttaagcacc cccccgccct ccgtggaggc
     1981 gggggtttgg tcggcgggtg gtaactggcg ggccgctgac tcgggcgggt cgcgcgcccc
     2041 agagtgtgac cttttcggtc tgctcgcaga cccccgggcg gcgccgccgc ggcggcgacg
     2101 ggctcgctgg gtcctaggct ccatggggac cgtatacgtg gacaggctct ggagcatccg
     2161 cacgactgcg gtgatattac cggagacctt ctgcgggacg agccgggtca cgcggctgac
     2221 gcggagcgtc cgttgggcga caaacaccag gacggggcac aggtacacta tcttgtcacc
     2281 cggaggcgcg agggactgca ggagcttcag ggagtggcgc agctgcttca tccccgtggc
     2341 ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc
     2401 tatgatgaca caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca
     2461 gtcggggcgg cgcggtccca ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa
     2521 caccgagcga ccctgcagcg acccgcttaa aagcttggca ttccggtact gttggtaaag
     2581 ccaccatggc cgatgctaag aacattaaga agggccctgc tcccttctac cctctggagg
     2641 atggcaccgc tggcgagcag ctgcacaagg ccatgaagag gtatgccctg gtgcctggca
     2701 ccattgcctt caccgatgcc cacattgagg tggacatcac ctatgccgag tacttcgaga
     2761 tgtctgtgcg cctggccgag gccatgaaga ggtacggcct gaacaccaac caccgcatcg
     2821 tggtgtgctc tgagaactct ctgcagttct tcatgccagt gctgggcgcc ctgttcatcg
     2881 gagtggccgt ggcccctgct aacgacattt acaacgagcg cgagctgctg aacagcatgg
     2941 gcatttctca gcctaccgtg gtgttcgtgt ctaagaaggg cctgcagaag atcctgaacg
     3001 tgcagaagaa gctgcctatc atccagaaga tcatcatcat ggactctaag accgactacc
     3061 agggcttcca gagcatgtac acattcgtga catctcatct gcctcctggc ttcaacgagt
     3121 acgacttcgt gccagagtct ttcgacaggg acaaaaccat tgccctgatc atgaacagct
     3181 ctgggtctac cggcctgcct aagggcgtgg ccctgcctca tcgcaccgcc tgtgtgcgct
     3241 tctctcacgc ccgcgaccct attttcggca accagatcat ccccgacacc gctattctga
     3301 gcgtggtgcc attccaccac ggcttcggca tgttcaccac cctgggctac ctgatttgcg
     3361 gctttcgggt ggtgctgatg taccgcttcg aggaggagct gttcctgcgc agcctgcaag
     3421 actacaaaat tcagtctgcc ctgctggtgc caaccctgtt cagcttcttc gctaagagca
     3481 ccctgatcga caagtacgac ctgtctaacc tgcacgagat tgcctctggc ggcgccccac
     3541 tgtctaagga ggtgggcgaa gccgtggcca agcgctttca tctgccaggc atccgccagg
     3601 gctacggcct gaccgagaca accagcgcca ttctgattac cccagagggc gacgacaagc
     3661 ctggcgccgt gggcaaggtg gtgccattct tcgaggccaa ggtggtggac ctggacaccg
     3721 gcaagaccct gggagtgaac cagcgcggcg agctgtgtgt gcgcggccct atgattatgt
     3781 ccggctacgt gaataaccct gaggccacaa acgccctgat cgacaaggac ggctggctgc
     3841 actctggcga cattgcctac tgggacgagg acgagcactt cttcatcgtg gaccgcctga
     3901 agtctctgat caagtacaag ggctaccagg tggccccagc cgagctggag tctatcctgc
     3961 tgcagcaccc taacattttc gacgccggag tggccggcct gcccgacgac gatgccggcg
     4021 agctgcctgc cgccgtcgtc gtgctggaac acggcaagac catgaccgag aaggagatcg
     4081 tggactatgt ggccagccag gtgacaaccg ccaagaagct gcgcggcgga gtggtgttcg
     4141 tggacgaggt gcccaagggc ctgaccggca agctggacgc ccgcaagatc cgcgagatcc
     4201 tgatcaaggc taagaaaggc ggcaagatcg ccgtgtaata attctagagt cggggcggcc
     4261 ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     4321 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4381 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4441 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg
     4501 ataaggatcc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4561 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4621 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4681 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4741 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4801 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4861 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4921 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4981 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5041 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5101 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5161 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5221 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5281 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5341 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5401 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5461 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5521 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5581 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5641 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5701 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5761 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5821 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5881 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5941 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6001 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6061 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6121 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6181 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6241 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6301 gctggccttt tgctcacatg gctcgac
//
