LOCUS       sequence-168349-        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"
     polyA_signal    39..160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(50..69)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     104..123
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(408..425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(579..1167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(659..678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1338..2129,2130..2198))
                     /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     1961..1980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2199..2303)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2330..2785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2417..2436)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2627..2648
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2916..2964
                     /note="synthetic polyadenylation signal"
     misc_feature    2978..3069
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3018..3037
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      3341..3350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3347..4999
                     /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(3413..3431)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
       61 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
      121 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
      181 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
      241 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag
      301 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
      361 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
      421 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
      481 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
      541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     1081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     1141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     1201 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     1261 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     1321 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     1381 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     1441 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1501 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1561 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1621 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1681 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1741 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1801 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1861 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1921 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1981 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     2041 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     2101 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     2161 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     2221 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     2281 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     2341 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2401 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2461 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2521 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2581 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2641 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2701 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2761 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
     2821 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
     2881 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
     2941 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca
     3001 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa
     3061 catttctcta tcgataggta ccgagctctt acgcgtccat gcatgctatg ggaattccca
     3121 tcagacgtct ggctcgagat ctgcgatcga gctcgcccgg ggatcgagcg cagcggtata
     3181 aaagggcgcg gggtggctga gagcatcagt tgtgaatgaa tgttcgagcc gagcagacgt
     3241 gccgctgcct tcgttaatat cctttgaata agccaacttt gaatcacaag acgcatacca
     3301 aaccctgcag gaagcttggc attccggtac tgttggtaaa gccaccatgg aagacgccaa
     3361 aaacataaag aaaggcccgg cgccattcta tccgctggaa gatggaaccg ctggagagca
     3421 actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc
     3481 acatatcgag gtggacatca cttacgctga gtacttcgaa atgtccgttc ggttggcaga
     3541 agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc
     3601 tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc
     3661 gaacgacatt tataatgaac gtgaattgct caacagtatg ggcatttcgc agcctaccgt
     3721 ggtgttcgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat
     3781 catccaaaaa attattatca tggattctaa aacggattac cagggatttc agtcgatgta
     3841 cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg tgccagagtc
     3901 cttcgatagg gacaagacaa ttgcactgat catgaactcc tctggatcta ctggtctgcc
     3961 taaaggtgtc gctctgcctc atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc
     4021 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     4081 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     4141 gtatagattt gaagaagagc tgtttctgag gagccttcag gattacaaga ttcaaagtgc
     4201 gctgctggtg ccaaccctat tctccttctt cgccaaaagc actctgattg acaaatacga
     4261 tttatctaat ttacacgaaa ttgcttctgg tggcgctccc ctctctaagg aagtcgggga
     4321 agcggttgcc aagaggttcc atctgccagg tatcaggcaa ggatatgggc tcactgagac
     4381 tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt
     4441 tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa
     4501 tcaaagaggc gaactgtgtg tgagaggtcc tatgattatg tccggttatg taaacaatcc
     4561 ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag acatagctta
     4621 ctgggacgaa gacgaacact tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa
     4681 aggctatcag gtggctcccg ctgaattgga atccatcttg ctccaacacc ccaacatctt
     4741 cgacgcaggt gtcgcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt
     4801 tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca
     4861 agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg
     4921 tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg
     4981 cggaaagatc gccgtgtaat t
//
