LOCUS       sequence-471816-        4915 bp    DNA     circular SYN 28-MAY-2025
DEFINITION  Express SBE response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4915)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4915)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 28, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pNL [NlucP/SBE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        119..150
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      177..186
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             183..695
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             699..818
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    870..991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(881..900)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     935..954
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1185..1542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1185..1376
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1208..1228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1393..1528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1455..1474
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1573..2610
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2589..2597
                     /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    2634..2682
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2838..2855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3009..3597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3089..3108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3797..4588,4589..4657))
                     /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    4762..4810
                     /note="synthetic polyadenylation signal"
     misc_feature    4824..4915
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4864..4883
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcagta tgtctagact gaagtatgtc tagactgaag
       61 tatgtctaga ctgactcgag gatatcaaga tctggcctcg gcggccaagc ttagacacta
      121 gagggtatat aatggaagct cgacttccag cttggcaatc cggtactgtt ggtaaagcca
      181 ccatggtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
      241 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
      301 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
      361 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
      421 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
      481 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
      541 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
      601 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
      661 tgaccggctg gcggctgtgc gaacgcattc tggcgaattc tcacggcttt ccgcctgagg
      721 ttgaagagca agccgccggt acattgccta tgtcctgcgc acaagaaagc ggtatggacc
      781 ggcacccagc cgcttgtgct tcagctcgca tcaacgtcta aggccgcgac tctagagtcg
      841 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
      901 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      961 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1021 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1081 taaaatcgat aaggatccgt ttgcgtattg ggcgctcttc cgctgatctg cgcagcacca
     1141 tggcctgaaa taacctctga aagaggaact tggttagcta ccttctgagg cggaaagaac
     1201 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     1261 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     1321 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     1381 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     1441 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     1501 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga ttcttctgac
     1561 actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa atttctcatc
     1621 gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga gagccgagcc
     1681 ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg cgctgatggc
     1741 ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc catccccgaa
     1801 gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag acgcgcccaa
     1861 ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca gcctgtcgcc
     1921 gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt cggcccattc
     1981 gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc cattgctgat
     2041 ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag cgtagctcaa
     2101 gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca cctcgtccat
     2161 gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc cgtaatcgac
     2221 tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt cttctggcgg
     2281 ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca tcccgagctg
     2341 gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca gctctaccag
     2401 agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg cgatgccatc
     2461 gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag cgcagccgta
     2521 tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc cagtacacga
     2581 ccgcgcgcta aggaggtagg tcgagtttaa actctagaac cggtcatggc cgcaataaaa
     2641 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgttcgaact agatgctgtc
     2701 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     2761 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     2821 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     2881 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     2941 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3001 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3061 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3121 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3181 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3241 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3301 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3361 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3421 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3481 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3541 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3601 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3661 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3721 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcgg ccgcaaatgc
     3781 taaaccactg cagtggttac cagtgcttga tcagtgaggc accgatctca gcgatctgcc
     3841 tatttcgttc gtccatagtg gcctgactcc ccgtcgtgta gatcactacg attcgtgagg
     3901 gcttaccatc aggccccagc gcagcaatga tgccgcgaga gccgcgttca ccggcccccg
     3961 atttgtcagc aatgaaccag ccagcaggga gggccgagcg aagaagtggt cctgctactt
     4021 tgtccgcctc catccagtct atgagctgct gtcgtgatgc tagagtaaga agttcgccag
     4081 tgagtagttt ccgaagagtt gtggccattg ctactggcat cgtggtatca cgctcgtcgt
     4141 tcggtatggc ttcgttcaac tctggttccc agcggtcaag ccgggtcaca tgatcaccca
     4201 tattatgaag aaatgcagtc agctccttag ggcctccgat cgttgtcaga agtaagttgg
     4261 ccgcggtgtt gtcgctcatg gtaatggcag cactacacaa ttctcttacc gtcatgccat
     4321 ccgtaagatg cttttccgtg accggcgagt actcaaccaa gtcgttttgt gagtagtgta
     4381 tacggcgacc aagctgctct tgcccggcgt ctatacggga caacaccgcg ccacatagca
     4441 gtactttgaa agtgctcatc atcgggaatc gttcttcggg gcggaaagac tcaaggatct
     4501 tgccgctatt gagatccagt tcgatatagc ccactcttgc acccagttga tcttcagcat
     4561 cttttacttt caccagcgtt tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga
     4621 agggaatgag tgcgacacga aaatgttgga tgctcatact cgtccttttt caatattatt
     4681 gaagcattta tcagggttac tagtacgtct ctcaaggata agtaagtaat attaaggtac
     4741 gggaggtatt ggacaggccg caataaaata tctttatttt cattacatct gtgtgttggt
     4801 tttttgtgtg aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac
     4861 aaactagcaa aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctct
//
