LOCUS       sequence-471836-        4975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SRF response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4975)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4975)
  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            "Unpublished"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: pNL [NlucP/SRF/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        179..210
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      237..246
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             243..755
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             759..878
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    930..1051
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(941..960)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     995..1014
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1245..1602
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1245..1436
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1268..1288)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1453..1588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1515..1534
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1633..2670
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2649..2657
                     /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    2694..2742
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2898..2915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3069..3657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3149..3168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3857..4648,4649..4717))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     polyA_signal    4822..4870
                     /note="synthetic polyadenylation signal"
     misc_feature    4884..4975
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4924..4943
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcagtatgtc catattagga catctaccat
       61 gtccatatta ggacatctac tatgtccata ttaggacatc ttgtatgtcc atattaggac
      121 atctaaaatg tccatattag gacatctaga tctggcctcg gcggccaagc ttagacacta
      181 gagggtatat aatggaagct cgacttccag cttggcaatc cggtactgtt ggtaaagcca
      241 ccatggtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
      301 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
      361 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
      421 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
      481 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
      541 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
      601 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
      661 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
      721 tgaccggctg gcggctgtgc gaacgcattc tggcgaattc tcacggcttt ccgcctgagg
      781 ttgaagagca agccgccggt acattgccta tgtcctgcgc acaagaaagc ggtatggacc
      841 ggcacccagc cgcttgtgct tcagctcgca tcaacgtcta aggccgcgac tctagagtcg
      901 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
      961 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     1021 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1081 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1141 taaaatcgat aaggatccgt ttgcgtattg ggcgctcttc cgctgatctg cgcagcacca
     1201 tggcctgaaa taacctctga aagaggaact tggttagcta ccttctgagg cggaaagaac
     1261 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     1321 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     1381 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     1441 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     1501 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     1561 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga ttcttctgac
     1621 actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa atttctcatc
     1681 gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga gagccgagcc
     1741 ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg cgctgatggc
     1801 ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc catccccgaa
     1861 gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag acgcgcccaa
     1921 ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca gcctgtcgcc
     1981 gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt cggcccattc
     2041 gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc cattgctgat
     2101 ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag cgtagctcaa
     2161 gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca cctcgtccat
     2221 gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc cgtaatcgac
     2281 tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt cttctggcgg
     2341 ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca tcccgagctg
     2401 gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca gctctaccag
     2461 agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg cgatgccatc
     2521 gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag cgcagccgta
     2581 tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc cagtacacga
     2641 ccgcgcgcta aggaggtagg tcgagtttaa actctagaac cggtcatggc cgcaataaaa
     2701 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgttcgaact agatgctgtc
     2761 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     2821 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     2881 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2941 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3001 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3061 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3121 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3181 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3241 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3301 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3361 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3421 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3481 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3541 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3601 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3661 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3721 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3781 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcgg ccgcaaatgc
     3841 taaaccactg cagtggttac cagtgcttga tcagtgaggc accgatctca gcgatctgcc
     3901 tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta gatcactacg attcgtgagg
     3961 gcttaccatc aggccccagc gcagcaatga tgccgcgaga gccgcgttca ccggcccccg
     4021 atttgtcagc aatgaaccag ccagcaggga gggccgagcg aagaagtggt cctgctactt
     4081 tgtccgcctc catccagtct atgagctgct gtcgtgatgc tagagtaaga agttcgccag
     4141 tgagtagttt ccgaagagtt gtggccattg ctactggcat cgtggtatca cgctcgtcgt
     4201 tcggtatggc ttcgttcaac tctggttccc agcggtcaag ccgggtcaca tgatcaccca
     4261 tattatgaag aaatgcagtc agctccttag ggcctccgat cgttgtcaga agtaagttgg
     4321 ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa ttctcttacc gtcatgccat
     4381 ccgtaagatg cttttccgtg accggcgagt actcaaccaa gtcgttttgt gagtagtgta
     4441 tacggcgacc aagctgctct tgcccggcgt ctatacggga caacaccgcg ccacatagca
     4501 gtactttgaa agtgctcatc atcgggaatc gttcttcggg gcggaaagac tcaaggatct
     4561 tgccgctatt gagatccagt tcgatatagc ccactcttgc acccagttga tcttcagcat
     4621 cttttacttt caccagcgtt tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga
     4681 agggaatgag tgcgacacga aaatgttgga tgctcatact cgtccttttt caatattatt
     4741 gaagcattta tcagggttac tagtacgtct ctcaaggata agtaagtaat attaaggtac
     4801 gggaggtatt ggacaggccg caataaaata tctttatttt cattacatct gtgtgttggt
     4861 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     4921 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctct
//
