LOCUS       sequence-472460-        7871 bp    DNA     circular SYN 13-MAY-2025
DEFINITION  Express -30F DEVD for Caspase 3/7 assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7871)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7871)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGloSensor -30F DEVD Caspase Vector
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2836..2854)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2891..3012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2902..2921)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2956..2975
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3257..3614
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3257..3448
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3280..3300)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3465..3600
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3659..3668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3665..4597
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4649..5440
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4700..4719)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5310..5329
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5504..5552
                     /note="synthetic polyadenylation signal"
     promoter        5708..5812
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5813..5881,5882..6673)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6031..6050)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6831..7419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6911..6930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7535..7818
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag gtttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggaattttaa ctttactaag gagcgatcgc
     1081 catgccggga tcaactggta aaatagtacc attacacgct gttaaagttg tcgatcctac
     1141 aacaggaaaa attttggggc caaatgaacc tggagaattg tattttaaag gcccgatgat
     1201 aatgaagggt tattataata atgaagaagc tactaaagca attattgata atgacggatg
     1261 gttgcgctct ggtgatattg cttattatga caatgatggc catttttata ttgtggacag
     1321 gctgaagtca ctgattaaat ataaaggtta tcaggttgca cctgctgaaa ttgagggaat
     1381 actcttacaa catccgtata ttgttgatgc cggcgttact ggtataccgg atgaagccgc
     1441 gggcgagctt ccagctgcag gtgttgtagt acagactgga aaatatctaa acgaacaaat
     1501 cgtacaagat tatgttgcca gtcaagtttc aatagccaaa tggctacgtg gtggggtgaa
     1561 atttttggat gaaattccca aaggatcaac tggaaaaatt gacagaaaag tgttaagaca
     1621 aatgttagaa aaacacacca atggatccga cgaggtggac ggaagcttga aaaacatcct
     1681 gtatggtccg gaaccgttct acccactgga agatggtacc gctggtgaac agatgtttga
     1741 cgcattatct cgttatgcag ctattccggg ctgcatagca ttgacaaatg ctcatacaaa
     1801 agaaaatgtt ttatatgaag agtttctgaa actgtcgtgt cgtttagcgg aaagttttaa
     1861 aaagtatgga ttaaaacaaa acgacacaat agcggtgtgt agcgaaaata gtctgcaatt
     1921 tttccttcct gtaattgcat cattgtatct tggaataatt gtggcacctg ttaacgataa
     1981 atacattgaa cgtgaattaa tacacagtct tggtattgta aaaccacgca tagttttttg
     2041 ctccaagaat acttttcaaa aagtactgaa tgtaaaatct aaattaaaat ctattgaaac
     2101 tattattata ttagacttaa atgaagactt aggaggttat caatgcctca acaactttat
     2161 ttctcaaaat tccgatagta atctggacgt aaaaaaattt aaaccctatt cttttaatcg
     2221 agacgatcag gttgcgtcga ttatgttttc ttctggtaca actggtctgc cgaagggagt
     2281 catgctaact cacaagaata ttgttgcacg attttctatt gcaaaagatc ctacttttgg
     2341 taacgcaatt aatcccacgt cagcaatttt aacggtaata cctttccacc atggttttgg
     2401 tatgatgacc acattaggat actttacttg tggattccga gttgttctaa tgcacacgtt
     2461 tgaagaaaaa ctatttctac aatcattaca agattataaa gtggaaagta ctttacttgt
     2521 accaacatta atggcatttc ttgcaaaaag tgcattagtt gaaaagtacg atttatcgca
     2581 cttaaaagaa attgcatctg gtggcgcacc tttatcaaaa gaaattgggg agatggtgaa
     2641 aaaacggttt aaattaaact ttgtcaggca agggtatgga ttaacagaaa ccacttcggc
     2701 tgttttaatt acaccgaaag gtgtttaaac gaattcgggc tcggtacccg gggttcctct
     2761 agagtcgacc tgcaggcatg caagctgatc cggctgctaa caaagcccga aaggaagctg
     2821 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcg gccgcttcga
     2881 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2941 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3001 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     3061 tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa
     3121 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3181 tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa
     3241 cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3301 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3361 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3421 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3481 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3541 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3601 taggcttttg caaaaaggtt gattcttctg acacaacagt ctcgaaccaa aggctggagc
     3661 caccatggct tccaaggtgt acgaccccga gcaacgcaaa cgcatgatca ctgggcctca
     3721 gtggtgggct cgctgcaagc aaatgaacgt gctggactcc ttcatcaact actatgattc
     3781 cgagaagcac gccgagaacg ccgtgatttt tctgcatggt aacgcagcct ccagctacct
     3841 gtggaggcac gtcgtgcctc acatcgagcc cgtggctaga tgcatcatcc ctgatctgat
     3901 cggaatgggt aagtccggca agagcgggaa tggctcatat cgcctcctgg atcactacaa
     3961 gtacctcacc gcttggttcg agctgctgaa ccttccaaag aaaatcatct ttgtgggcca
     4021 cgactggggg gcttgtctgg cctttcacta ctcctacgag caccaagaca agatcaaggc
     4081 catcgtccat gctgagagtg tcgtggacgt gatcgagtcc tgggacgagt ggcctgacat
     4141 cgaggaggat atcgccctga tcaagagcga agagggcgag aaaatggtgc ttgagaataa
     4201 cttcttcgtc gagaccatgc tcccaagcaa gatcatgcgg aaactggagc ctgaggagtt
     4261 cgctgcctac ctggagccat tcaaggagaa gggcgaggtt agacggccta ccctctcctg
     4321 gcctcgcgag atccctctcg ttaagggagg caagcccgac gtcgtccaga ttgtccgcaa
     4381 ctacaacgcc taccttcggg ccagcgacga tctgcctaag atgttcatcg agtccgaccc
     4441 tgggttcttt tccaacgcta ttgtcgaggg agctaagaag ttccctaaca ccgagttcgt
     4501 gaaggtgaag ggcctccact tcagccagga ggacgctcca gatgaaatgg gtaagtacat
     4561 caagagcttc gtggagcgcg tgctgaagaa cgagcagacc ggtggtggga gcggaggtgg
     4621 cggatcaggt ggcggaggct ccggagggat tgaacaagat ggattgcacg caggttctcc
     4681 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4741 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4801 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4861 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4921 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4981 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     5041 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     5101 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     5161 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     5221 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     5281 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     5341 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     5401 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     5461 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     5521 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataaggt tcctctttgc
     5581 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
     5641 tttctgcgga ctggctttct acgtaatggt ttcttagacg tcaggtggca cttttcgggg
     5701 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5761 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5821 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5881 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5941 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6001 ttttccaatg atgagcactt tcaaagttct gctatgtggc gcggtattat cccgtattga
     6061 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     6121 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     6181 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacta tcggaggacc
     6241 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     6301 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     6361 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6421 acaattaata gactggatgg aggcggataa agttgcagga tcacttctgc gctcggccct
     6481 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6541 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6601 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6661 taagcattgg taattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     6721 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6781 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6841 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6901 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6961 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7021 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7081 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7141 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7201 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7261 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7321 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7381 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     7441 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7501 tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac gcacgctgga
     7561 tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga ttaagtgggg
     7621 gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat ttatcaggcg
     7681 cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct gccgtgatcg
     7741 cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc acttactgtc
     7801 tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc cggcgtagag
     7861 gatcgagatc t
//
