LOCUS       sequence-127081-        5488 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 5488)
  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 5488)
  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_530_S2-1-CGCG_dCP
FEATURES             Location/Qualifiers
     source          1..5488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(10..801,802..870))
                     /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     633..652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(871..975)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1002..1457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1089..1108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1299..1320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1588..1636
                     /note="synthetic polyadenylation signal"
     misc_feature    1650..1741
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1690..1709
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    1755..1779
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2285..2309)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2500..2509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2506..4158
                     /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(2572..2590)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4199..4320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4210..4229)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4264..4283
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4568..4585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4739..5327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4819..4838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
       61 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
      121 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
      181 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
      241 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
      301 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
      361 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
      421 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
      481 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
      541 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
      601 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
      661 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
      721 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
      781 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
      841 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
      901 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
      961 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     1021 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     1081 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1141 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1201 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1261 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1321 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     1381 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     1441 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     1501 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
     1561 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     1621 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     1681 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     1741 tatcgatagg taccacaagt ttgtacaaaa aagcaggctc cgaattcgcc ctttgcacaa
     1801 tttctgtgaa gtgacagcca ctgagaaata gtgtcaagta gctctcgtcc gctaatatca
     1861 atttagacga tatacatacg gccagatctg gcacgcagtc tgcattccca atgatgcggg
     1921 atggatttgg attgtggggc ggaatctcta agtgcgataa gatgcggaga attactcgga
     1981 ggacagtact ccgtggctcc gttttgttaa atatactatg tgtgtatgtg cgtagtacag
     2041 cgacttgtac ggagtacata aacatatgca tatctgcggt ttagttgtcg acattatctt
     2101 ttgttcgtcg tttgaaaaat gtgtgtagtt gttggggtac ggagtactgt cctccgagat
     2161 tgccgggcta cagggtgctt tgagtctttg tttttggatt tcgttttcga tgtttagcta
     2221 cgaaagccaa gcgttgtgct attttcgaat tttgtatttt ataacccggg caagggcgaa
     2281 ttcgacccag ctttcttgta caaagtggta gatctgcgat cgagctcgcc cggggatcga
     2341 gcgcagcggt ataaaagggc gcggggtggc tgagagcatc agttgtgaat gaatgttcga
     2401 gccgagcaga cgtgccgctg ccttcgttaa tatcctttga ataagccaac tttgaatcac
     2461 aagacgcata ccaaacggca ttccggtact gttggtaaag ccaccatgga agacgccaaa
     2521 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
     2581 ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt tacagatgca
     2641 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
     2701 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
     2761 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     2821 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
     2881 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
     2941 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     3001 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
     3061 ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac tggtctgcct
     3121 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
     3181 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     3241 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     3301 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     3361 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     3421 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     3481 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     3541 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     3601 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     3661 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     3721 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     3781 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     3841 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     3901 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     3961 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     4021 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     4081 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     4141 ggaaagatcg ccgtgtaatt ctagagtcgg ggcggccggc cgcttcgagc agacatgata
     4201 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     4261 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     4321 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     4381 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc
     4441 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     4501 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     4561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5161 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5461 caatctaaag tatatatgag taaacttg
//
