LOCUS       sequence-127114-        5413 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5413)
  AUTHORS   Stampfel G, Kazmar T, Frank O, Wienerroither S, Reiter F, Stark A
  TITLE     Transcriptional regulators form diverse groups with 
            context-dependent regulatory functions.
  JOURNAL   Nature. 2015 Nov 9. doi: 10.1038/nature15545.
  PUBMED    26550828
REFERENCE   2  (bases 1 to 5413)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2015 Nov 9. doi: 10.1038/nature15545."
COMMENT     Sequence Label: pSGE_892_Nhe2_EcR-to-HLH4C_RB
FEATURES             Location/Qualifiers
     source          1..5413
                     /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"
     protein_bind    3083..3107
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3538..3562)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3753..3762
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3759..5411
                     /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(3825..3843)
                     /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 ccacaagttt gtacaaaaaa gcaggcttcc tgcatcaaaa
     3121 acacctgcgc ccaggtccat tattcataaa aaaacacctg cgccaacaga gactactcgc
     3181 gaatccgaag acccaaaccg aatattacta tcactcattc gtaaaatcaa agttcacccc
     3241 aagaatcgac tcgattcaag tcactaacaa ggttcttaca ggccttcaaa aattagactc
     3301 atggattatg ggtagagccg ttgaacactt gaatgagtgt cttaataggg tggccacgtt
     3361 cgtaaaaaca cctgcgccta ttaggggacg gtaccctgga agaagctttc agtgcaaagg
     3421 acgagttaat agtttcaata cccggtcaat aataagcacc gacacgaaac aattataatg
     3481 attatacgtt cttggtctct gttaacctta atccagataa taggtcgcaa attaaagacc
     3541 cagctttctt gtacaaagtg gtagatctgc gatcgagctc gcccggggat cgagcgcagc
     3601 ggtataaaag ggcgcggggt ggctgagagc atcagttgtg aatgaatgtt cgagccgagc
     3661 agacgtgccg ctgccttcgt taatatcctt tgaataagcc aactttgaat cacaagacgc
     3721 ataccaaacg gcattccggt actgttggta aagccaccat ggaagacgcc aaaaacataa
     3781 agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata
     3841 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
     3901 aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga
     3961 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
     4021 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
     4081 tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg
     4141 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa
     4201 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     4261 tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata
     4321 gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg
     4381 tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg
     4441 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
     4501 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     4561 ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg
     4621 tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     4681 atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg
     4741 ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag
     4801 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     4861 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag
     4921 gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     4981 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     5041 aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc
     5101 aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag
     5161 gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     5221 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     5281 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     5341 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga
     5401 tcgccgtgta att
//
