LOCUS       sequence-127110-        5401 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 5401)
  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 5401)
  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_889_Nhe2_EcR-to-Ets96B_RB
FEATURES             Location/Qualifiers
     source          1..5401
                     /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(3526..3550)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3741..3750
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3747..5399
                     /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(3813..3831)
                     /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 tgcatcaccg
     3121 gaagtaccag gtccattatt cataaaccgg aagtacaaca gagactactc gcgaatccga
     3181 agacccaaac cgaatattac tatcactcat tcgtaaaatc aaagttcacc ccaagaatcg
     3241 actcgattca agtcactaac aaggttctta caggccttca aaaattagac tcatggatta
     3301 tgggtagagc cgttgaacac ttgaatgagt gtcttaatag ggtggccacg ttcgtaccgg
     3361 aagtactatt aggggacggt accctggaag aagctttcag tgcaaaggac gagttaatag
     3421 tttcaatacc cggtcaataa taagcaccga cacgaaacaa ttataatgat tatacgttct
     3481 tggtctctgt taaccttaat ccagataata ggtcgcaaat taaagaccca gctttcttgt
     3541 acaaagtggt agatctgcga tcgagctcgc ccggggatcg agcgcagcgg tataaaaggg
     3601 cgcggggtgg ctgagagcat cagttgtgaa tgaatgttcg agccgagcag acgtgccgct
     3661 gccttcgtta atatcctttg aataagccaa ctttgaatca caagacgcat accaaacggc
     3721 attccggtac tgttggtaaa gccaccatgg aagacgccaa aaacataaag aaaggcccgg
     3781 cgccattcta tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga
     3841 gatacgccct ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca
     3901 cttacgctga gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc
     3961 tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg
     4021 tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac
     4081 gtgaattgct caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg
     4141 ggttgcaaaa aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca
     4201 tggattctaa aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc
     4261 tacctcccgg ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa
     4321 ttgcactgat catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc
     4381 atagaactgc ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca
     4441 ttccggatac tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta
     4501 cactcggata tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc
     4561 tgtttctgag gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat
     4621 tctccttctt cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa
     4681 ttgcttctgg tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc
     4741 atctgccagg tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta
     4801 cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga
     4861 aggttgtgga tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg
     4921 tgagaggtcc tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga
     4981 ttgacaagga tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact
     5041 tcttcatcgt tgaccgcctg aagtctctga ttaagtacaa aggctatcag gtggctcccg
     5101 ctgaattgga atccatcttg ctccaacacc ccaacatctt cgacgcaggt gtcgcaggtc
     5161 ttcccgacga tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga
     5221 cgatgacgga aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt
     5281 tgcgcggagg agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg
     5341 caagaaaaat cagagagatc ctcataaagg ccaagaaggg cggaaagatc gccgtgtaat
     5401 t
//
