LOCUS       sequence-474931-        5276 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  Express TAF1-NanoLuc(R) Fusion Protein in Mammalian Cells under a 
            CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5276)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: NanoBRET Assay Vector, TAF1-NanoLuc
FEATURES             Location/Qualifiers
     source          1..5276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(72..92)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        247..550
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        670..688
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     670..687
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     polyA_signal    1920..2001
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1957..1976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2073..2430
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2073..2264
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2096..2116)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2281..2416
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2461..3498
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             3477..3485
                     /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    3522..3570
                     /note="synthetic polyadenylation signal"
     rep_origin      3696..4284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4185..4204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4442..5236)
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4553..4572)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5163..5182
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5268..5276,1..39))
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
ORIGIN
        1 tactatgccg atatactatg ccgatgatta attgtcaaca gtattagaat tcgatttgac
       61 ctcccaccgt acacgcctac cgcccatttg cgtcaacggg gcggggttat tacgacattt
      121 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
      181 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttggtgtact gccaaaaccg
      241 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtcccg
      301 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
      361 gggcggactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
      421 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
      481 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
      541 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
      601 ttaataacta gtggcgcgcc agggaggcct atataagcag agctggttta gtgaaccgtc
      661 agatcgccca tttaggtgac actatagaat acaagctact tgttcttttt gcagctagcg
      721 atcgccatga cgacccccaa taaaggaaac aaggccttga aggtgaagcg ggagccgggt
      781 gagaatggca ccagcctgac ggatgaggag ctggtgacca tgtcggtgcg ggagctgaac
      841 cagcacctgc ggggcctgtc caaggaggag atcgtccagc tgaagcagcg ccggcgcacg
      901 ctcaagaacc gcggctacgc tgccagctgc cgcgtgaagc gggtgacgca gaaggaggag
      961 ctggagaagc agaaggcgga gctgcagcag gaggtggaga agctggcctc agagaacgcc
     1021 agcatgaagc tggagctcga cgcgctgcgc tccaagtacg aggcgctgca gaccttcgcc
     1081 cggacggtgg cccgcagccc cgtggcgcca gcccggggcc cccttgccgc cggcctgggg
     1141 cccctcgtcc caggcaaggt ggccgccacc agcgtcatca caatagtaaa gtccaagacg
     1201 gatgcccgat cggtttctca aggcagttca ggtggtggcg ggagcggagg tggaggctcg
     1261 agcggtgtct tcacactcga agatttcgtt ggggactggg aacagacagc cgcctacaac
     1321 ctggaccaag tccttgaaca gggaggtgtg tccagtttgc tgcagaatct cgccgtgtcc
     1381 gtaactccga tccaaaggat tgtccggagc ggtgaaaatg ccctgaagat cgacatccat
     1441 gtcatcatcc cgtatgaagg tctgagcgcc gaccaaatgg cccagatcga agaggtgttt
     1501 aaggtggtgt accctgtgga tgatcatcac tttaaggtga tcctgcccta tggcacactg
     1561 gtaatcgacg gggttacgcc gaacatgctg aactatttcg gacggccgta tgaaggcatc
     1621 gccgtgttcg acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt
     1681 atcgacgagc gcctgatcac ccccgacggc tccatgctgt tccgagtaac catcaacagc
     1741 taaggccgcg actctagaaa cacctaggct ctcgtacgaa gtcagctgat aacgtcgacg
     1801 tattgcggcc gcggatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     1861 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     1921 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1981 ataaagcatt tttttcactg ccgacgtcac gcgtcttgcg cagcaccatg gcctgaaata
     2041 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     2101 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2161 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2221 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2281 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2341 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2401 ggcttttttg gaggcctagg cttttgcaaa aagctcgatt cttctgacac tagcgccacc
     2461 atgaagaagc ccgaactcac cgctaccagc gttgaaaaat ttctcatcga gaagttcgac
     2521 agtgtgagcg acctgatgca gttgtcggag ggcgaagaga gccgagcctt cagcttcgat
     2581 gtcggcggac gcggctatgt actgcgggtg aatagctgcg ctgatggctt ctacaaagac
     2641 cgctacgtgt accgccactt cgccagcgct gcactaccca tccccgaagt gttggacatc
     2701 ggcgagttca gcgagagcct gacatactgc atcagtagac gcgcccaagg cgttactctc
     2761 caagacctcc ccgaaacaga gctgcctgct gtgttacagc ctgtcgccga agctatggat
     2821 gctattgccg ccgccgacct cagtcaaacc agcggcttcg gcccattcgg gccccaaggc
     2881 atcggccagt acacaacctg gcgggatttc atttgcgcca ttgctgatcc ccatgtctac
     2941 cactggcaga ccgtgatgga cgacaccgtg tccgccagcg tagctcaagc cctggacgaa
     3001 ctgatgctgt gggccgaaga ctgtcccgag gtgcgccacc tcgtccatgc cgacttcggc
     3061 agcaacaacg tcctgaccga caacggccgc atcaccgccg taatcgactg gtccgaagct
     3121 atgttcgggg acagtcagta cgaggtggcc aacatcttct tctggcggcc ctggctggct
     3181 tgcatggagc agcagactcg ctacttcgag cgccggcatc ccgagctggc cggcagccct
     3241 cgtctgcgag cctacatgct gcgcatcggc ctggatcagc tctaccagag cctcgtggac
     3301 ggcaacttcg acgatgctgc ctgggctcaa ggccgctgcg atgccatcgt ccgcagcggg
     3361 gccggcaccg tcggtcgcac acaaatcgct cgccggagcg cagccgtatg gaccgacggc
     3421 tgcgtcgagg tgctggccga cagcggcaac cgccggccca gtacacgacc gcgcgctaag
     3481 gaggtaggtc gagtttagac tctagccatc acgatggccg caataaaata tctttatttt
     3541 cattacatct gtgtgttggt tttttgtgtg acgattccgg actataaaag gatctaggtg
     3601 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3661 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgaa atcctttttt tctgcgcgta
     3721 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3781 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3841 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3901 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3961 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4021 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4081 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4141 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4201 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4261 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4321 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4381 cgtattaccg cctttgagtg agctgatacc ggttgggcgt cgcttggtcg gtcatttcga
     4441 atcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     4501 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     4561 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     4621 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     4681 cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg
     4741 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     4801 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     4861 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     4921 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     4981 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     5041 gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     5101 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     5161 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     5221 catcttgttc aatcatgatc tgcctcctgg gtttagttcc tcaccttgtc gtatta
//
