LOCUS       sequence-471830-        4947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CRE response element in luciferase reporter assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4947)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4947)
  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            "Unpublished"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/CRE/Hygro] Vector
FEATURES             Location/Qualifiers
     source          1..4947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        151..182
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      209..218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             215..727
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             731..850
                     /codon_start=1
                     /product="PEST degradation sequence from mouse ornithine 
                     decarboxylase"
                     /label=hPEST
                     /note="human codon-optimized"
                     /translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
     polyA_signal    902..1023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(913..932)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     967..986
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1217..1574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1217..1408
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1240..1260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1425..1560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1487..1506
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1605..2642
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2621..2629
                     /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    2666..2714
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(2870..2887)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3041..3629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3121..3140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3829..4620,4621..4689))
                     /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    4794..4842
                     /note="synthetic polyadenylation signal"
     misc_feature    4856..4947
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4896..4915
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggcctaactg gccggtacct gagctcgcta gcgcaccaga cagtgacgtc agctgccaga
       61 tcccatggcc gtcatactgt gacgtctttc agacacccca ttgacgtcaa tgggagaaca
      121 gatctggcct cggcggccaa gcttagacac tagagggtat ataatggaag ctcgacttcc
      181 agcttggcaa tccggtactg ttggtaaagc caccatggtc ttcacactcg aagatttcgt
      241 tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac agggaggtgt
      301 gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga ttgtcctgag
      361 cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg
      421 cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg atgatcatca
      481 ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc cgaacatgat
      541 cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa agatcactgt
      601 aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca accccgacgg
      661 ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat
      721 tctggcgaat tctcacggct ttccgcctga ggttgaagag caagccgccg gtacattgcc
      781 tatgtcctgc gcacaagaaa gcggtatgga ccggcaccca gccgcttgtg cttcagctcg
      841 catcaacgtc taaggccgcg actctagagt cggggcggcc ggccgcttcg agcagacatg
      901 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
      961 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1021 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1081 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtttgcgtat
     1141 tgggcgctct tccgctgatc tgcgcagcac catggcctga aataacctct gaaagaggaa
     1201 cttggttagc taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     1261 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1321 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1381 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     1441 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     1501 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     1561 taggcttttg caaaaagctc gattcttctg acactagcgc caccatgaag aagcccgaac
     1621 tcaccgctac cagcgttgaa aaatttctca tcgagaagtt cgacagtgtg agcgacctga
     1681 tgcagttgtc ggagggcgaa gagagccgag ccttcagctt cgatgtcggc ggacgcggct
     1741 atgtactgcg ggtgaatagc tgcgctgatg gcttctacaa agaccgctac gtgtaccgcc
     1801 acttcgccag cgctgcacta cccatccccg aagtgttgga catcggcgag ttcagcgaga
     1861 gcctgacata ctgcatcagt agacgcgccc aaggcgttac tctccaagac ctccccgaaa
     1921 cagagctgcc tgctgtgtta cagcctgtcg ccgaagctat ggatgctatt gccgccgccg
     1981 acctcagtca aaccagcggc ttcggcccat tcgggcccca aggcatcggc cagtacacaa
     2041 cctggcggga tttcatttgc gccattgctg atccccatgt ctaccactgg cagaccgtga
     2101 tggacgacac cgtgtccgcc agcgtagctc aagccctgga cgaactgatg ctgtgggccg
     2161 aagactgtcc cgaggtgcgc cacctcgtcc atgccgactt cggcagcaac aacgtcctga
     2221 ccgacaacgg ccgcatcacc gccgtaatcg actggtccga agctatgttc ggggacagtc
     2281 agtacgaggt ggccaacatc ttcttctggc ggccctggct ggcttgcatg gagcagcaga
     2341 ctcgctactt cgagcgccgg catcccgagc tggccggcag ccctcgtctg cgagcctaca
     2401 tgctgcgcat cggcctggat cagctctacc agagcctcgt ggacggcaac ttcgacgatg
     2461 ctgcctgggc tcaaggccgc tgcgatgcca tcgtccgcag cggggccggc accgtcggtc
     2521 gcacacaaat cgctcgccgg agcgcagccg tatggaccga cggctgcgtc gaggtgctgg
     2581 ccgacagcgg caaccgccgg cccagtacac gaccgcgcgc taaggaggta ggtcgagttt
     2641 aaactctaga accggtcatg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     2701 tggttttttg tgtgttcgaa ctagatgctg tcgaccgatg cccttgagag ccttcaaccc
     2761 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     2821 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     2881 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     2941 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3481 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3781 agtaaacttg gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt accagtgctt
     3841 gatcagtgag gcaccgatct cagcgatctg cctatttcgt tcgtccatag tggcctgact
     3901 ccccgtcgtg tagatcacta cgattcgtga gggcttacca tcaggcccca gcgcagcaat
     3961 gatgccgcga gagccgcgtt caccggcccc cgatttgtca gcaatgaacc agccagcagg
     4021 gagggccgag cgaagaagtg gtcctgctac tttgtccgcc tccatccagt ctatgagctg
     4081 ctgtcgtgat gctagagtaa gaagttcgcc agtgagtagt ttccgaagag ttgtggccat
     4141 tgctactggc atcgtggtat cacgctcgtc gttcggtatg gcttcgttca actctggttc
     4201 ccagcggtca agccgggtca catgatcacc catattatga agaaatgcag tcagctcctt
     4261 agggcctccg atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc
     4321 agcactacac aattctctta ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga
     4381 gtactcaacc aagtcgtttt gtgagtagtg tatacggcga ccaagctgct cttgcccggc
     4441 gtctatacgg gacaacaccg cgccacatag cagtactttg aaagtgctca tcatcgggaa
     4501 tcgttcttcg gggcggaaag actcaaggat cttgccgcta ttgagatcca gttcgatata
     4561 gcccactctt gcacccagtt gatcttcagc atcttttact ttcaccagcg tttcggggtg
     4621 tgcaaaaaca ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg
     4681 gatgctcata ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt actagtacgt
     4741 ctctcaagga taagtaagta atattaaggt acgggaggta ttggacaggc cgcaataaaa
     4801 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat
     4861 acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg
     4921 caagtgcagg tgccagaaca tttctct
//
