LOCUS       sequence-173962-        5867 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Luciferase validation vector, where candidates are inserted at the 
            position of the core promoter. The candidates responsiveness to the 
            upstream enhancer is reflected by luciferase activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5867)
  AUTHORS   Arnold CD, Zabidi MA, Pagani M, Rath M, Schernhuber K, Kazmar T, 
            Stark A
  TITLE     Genome-wide assessment of sequence-intrinsic enhancer responsiveness
            at single-base-pair resolution.
  JOURNAL   Nat Biotechnol. 2016 Dec 26. doi: 10.1038/nbt.3739.
  PUBMED    28024147
REFERENCE   2  (bases 1 to 5867)
  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: "Nat 
            Biotechnol. 2016 Dec 26. doi: 10.1038/nbt.3739."
COMMENT     Sequence Label: pGL3_zfh1_CP-candidate_luc+
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      1065..1074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1071..2723
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1137..1155)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2764..2885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2775..2794)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2829..2848
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3133..3150)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3304..3892)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3384..3403)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4063..4854,4855..4923))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4686..4705
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4924..5028)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5055..5510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5142..5161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5352..5373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5641..5689
                     /note="synthetic polyadenylation signal"
     misc_feature    5703..5794
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5743..5762
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 aaaaaaaaaa aaaaaactaa attccaagtg agtagtttac tcttagtcac tatcgagtgt
       61 gtgctgtttg tttttcgaaa tatgcatatc ttagctactt gcgttgctgt tttttttttt
      121 tttttttttt tgacgatttg tcaacaggaa atatgcataa ggttggctcg ggatccggat
      181 cggatggcat ggggtttgat tggatgccgt acttggcacc gcactgccgt tatcggttgg
      241 ctcatttccc gttagccgcg aacaaagccc acgcattgtg ctacgaccgc cgtgatgtgt
      301 tcaaagatca agtgccccca gaaccatctc ctgccatccc tgccatccct gcccgcctgc
      361 atccatcgcg ggcattcaga ttccgactgc gcatgagcag cggtctccaa cacgtaagca
      421 agtggaaggc gagggataag aacaagtaac ttctctctta tcggacgctt ccaggaattt
      481 cgatcctcgc accgcattcc gcaggtttag tcactggatt ccacggcttt tcgagtcgat
      541 tgtcccaaga aacattattt tggacatgtg actcactgac tcaaagctat tatcacttct
      601 cggacatatg cgtgtatata aaatattttt aaacattatt aggtatacta taaactctgc
      661 ctcattcatg cctacagaac ttcagccttg acattctata tatagaaacc cttactacgt
      721 ctttactctt taaatagttc tttggatatt atattagcac attgatgtgc tttttttttg
      781 gatggttgtg tccaactcac ggtaccgagc tcttacgcgt ccatgcatgc tatgggaatt
      841 cccatcagac gtctggctcg agatctgcga tcgagctcgc ccggggatcg agcgcagcgg
      901 tataaaaggg cgcggggtgg ctgagagcat cagttgtgaa tgaatgttcg agccgagcag
      961 acgtgccgct gccttcgtta atatcctttg aataagccaa ctttgaatca caagacgcat
     1021 accaaaccct gcaggaagct tggcattccg gtactgttgg taaagccacc atggaagacg
     1081 ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga accgctggag
     1141 agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag
     1201 atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc gttcggttgg
     1261 cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa
     1321 actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc
     1381 ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt tcgcagccta
     1441 ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc
     1501 caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga
     1561 tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtgccag
     1621 agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga tctactggtc
     1681 tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg catgccagag
     1741 atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc
     1801 atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct
     1861 taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac aagattcaaa
     1921 gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg attgacaaat
     1981 acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct aaggaagtcg
     2041 gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat gggctcactg
     2101 agactacatc agctattctg attacacccg agggggatga taaaccgggc gcggtcggta
     2161 aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg
     2221 ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca
     2281 atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag
     2341 cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct ctgattaagt
     2401 acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa caccccaaca
     2461 tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg
     2521 ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca
     2581 gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga
     2641 aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga
     2701 agggcggaaa gatcgccgtg taattctaga gtcggggcgg ccggccgctt cgagcagaca
     2761 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2821 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2881 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2941 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg
     3001 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     3061 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     3121 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3181 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3241 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3301 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3361 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3421 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3481 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3541 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3601 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3661 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3721 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3781 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3841 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3901 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3961 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4021 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4081 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4141 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4201 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4261 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4321 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4381 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4441 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4501 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4561 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4621 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4681 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4741 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4801 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4861 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4921 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4981 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5041 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5101 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5161 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5221 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5281 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5341 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5401 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5461 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc
     5521 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5581 gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg gagcggccgc
     5641 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
     5701 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
     5761 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtacctta aaagtgccag
     5821 tgcttgtggc gtgcacggag aaaaaaacct tgaggtcata tttgaaa
//
