LOCUS       sequence-173946-        5001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Luciferase validation vector, where candidates are inserted at the 
            position of the core promoter. The candidates basal activity (no 
            enhancer in vector) is reflected by luciferase activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5001)
  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 5001)
  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_ctrl_CP-candidate_luc+
FEATURES             Location/Qualifiers
     source          1..5001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    263..354
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(295..314)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    368..416
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(547..1002)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(684..705)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     896..915
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1029..1133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1134..1202,1203..1994)
                     /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     complement(1352..1371)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2165..2753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2654..2673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2907..2924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3172..3293
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3209..3228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3263..3282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3334..4986)
                     /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     4902..4920
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      4983..4992
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agtaccggaa tgccaagctt cctgcagggt ttggtatgcg tcttgtgatt caaagttggc
       61 ttattcaaag gatattaacg aaggcagcgg cacgtctgct cggctcgaac attcattcac
      121 aactgatgct ctcagccacc ccgcgccctt ttataccgct gcgctcgatc cccgggcgag
      181 ctcgatcgca gatctcgagc cagacgtctg atgggaattc ccatagcatg catggacgcg
      241 taagagctcg gtacctatcg atagagaaat gttctggcac ctgcacttgc actggggaca
      301 gcctattttg ctagtttgtt ttgtttcgtt ttgttttgat ggagagcgta tgttagtact
      361 atcgattcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
      421 ccgctccaag tacctcccgt accttaatat tacttactta tcatggtagc ttgggctggc
      481 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      541 aatggcaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
      601 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      661 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      721 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      781 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      841 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      901 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      961 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     1021 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1081 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1141 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1201 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1261 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1321 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1381 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1441 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1501 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1561 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1621 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1681 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1741 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1801 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1861 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1921 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1981 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2041 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2101 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2161 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2281 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2521 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2581 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2641 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2701 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2761 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2821 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2881 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgct
     2941 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     3001 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     3061 cgacggatcc ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3121 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3181 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3241 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3301 tgctcgaagc ggccggccgc cccgactcta gaattacacg gcgatctttc cgcccttctt
     3361 ggcctttatg aggatctctc tgatttttct tgcgtcgagt tttccggtaa gacctttcgg
     3421 tacttcgtcc acaaacacaa ctcctccgcg caactttttc gcggttgtta cttgactggc
     3481 gacgtaatcc acgatctctt tttccgtcat cgtctttccg tgctccaaaa caacaacggc
     3541 ggcgggaagt tcaccggcgt catcgtcggg aagacctgcg acacctgcgt cgaagatgtt
     3601 ggggtgttgg agcaagatgg attccaattc agcgggagcc acctgatagc ctttgtactt
     3661 aatcagagac ttcaggcggt caacgatgaa gaagtgttcg tcttcgtccc agtaagctat
     3721 gtctccagaa tgtagccatc catccttgtc aatcaaggcg ttggtcgctt ccggattgtt
     3781 tacataaccg gacataatca taggacctct cacacacagt tcgcctcttt gattaacgcc
     3841 cagcgttttc ccggtatcca gatccacaac cttcgcttca aaaaatggaa caactttacc
     3901 gaccgcgccc ggtttatcat ccccctcggg tgtaatcaga atagctgatg tagtctcagt
     3961 gagcccatat ccttgcctga tacctggcag atggaacctc ttggcaaccg cttccccgac
     4021 ttccttagag aggggagcgc caccagaagc aatttcgtgt aaattagata aatcgtattt
     4081 gtcaatcaga gtgcttttgg cgaagaagga gaatagggtt ggcaccagca gcgcactttg
     4141 aatcttgtaa tcctgaaggc tcctcagaaa cagctcttct tcaaatctat acattaagac
     4201 gactcgaaat ccacatatca aatatccgag tgtagtaaac attccaaaac cgtgatggaa
     4261 tggaacaaca cttaaaatcg cagtatccgg aatgatttga ttgccaaaaa taggatctct
     4321 ggcatgcgag aatctcacgc aggcagttct atgaggcaga gcgacacctt taggcagacc
     4381 agtagatcca gaggagttca tgatcagtgc aattgtcttg tccctatcga aggactctgg
     4441 cacaaaatcg tattcattaa aaccgggagg tagatgagat gtgacgaacg tgtacatcga
     4501 ctgaaatccc tggtaatccg ttttagaatc catgataata attttttgga tgattgggag
     4561 ctttttttgc acgttcaaaa ttttttgcaa cccctttttg gaaacgaaca ccacggtagg
     4621 ctgcgaaatg cccatactgt tgagcaattc acgttcatta taaatgtcgt tcgcgggcgc
     4681 aactgcaact ccgataaata acgcgcccaa caccggcata aagaattgaa gagagttttc
     4741 actgcatacg acgattctgt gatttgtatt cagcccatat cgtttcatag cttctgccaa
     4801 ccgaacggac atttcgaagt actcagcgta agtgatgtcc acctcgatat gtgcatctgt
     4861 aaaagcaatt gttccaggaa ccagggcgta tctcttcata gccttatgca gttgctctcc
     4921 agcggttcca tcttccagcg gatagaatgg cgccgggcct ttctttatgt ttttggcgtc
     4981 ttccatggtg gctttaccaa c
//
