LOCUS       sequence-422217-        6291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrated transcriptional and translational control for stringently
            blue-light-activated gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6291)
  AUTHORS   Ranzani AT, Buchholz K, Blackholm M, Kopkin H, Moglich A
  TITLE     Induction of bacterial expression at the mRNA level by light.
  JOURNAL   Nucleic Acids Res. 2024 Aug 10:gkae678. doi: 10.1093/nar/gkae678.
  PUBMED    39126322
REFERENCE   2  (bases 1 to 6291)
  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: "Nucleic 
            Acids Res. 2024 Aug 10:gkae678. doi: 10.1093/nar/gkae678."
COMMENT     Sequence Label: pAurora2
FEATURES             Location/Qualifiers
     source          1..6291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     199..218
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1439..1468
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1521..1539)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1535..1582
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1636..1653)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             1797..2507
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             2508..2540
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      2588..2659
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2675..2702
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2878..2895
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2905..2922
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      3102..3188
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3280..3307
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3508..4299)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4300..4391)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4439..5177)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(5378..6256)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa
       61 ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga
      121 caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc
      181 acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg gatcctacct
      241 ttttaaaaaa ttcatttgct aaacgcttca aattctcgta taattacttt tctccatacc
      301 cgttttttgg gctaacccgc cgaaacctaa ggaaatacat agtatgaaag ttaatcgtcc
      361 ggcagaacgt gccagctttg gtagctttgt tctggatgca ggtagcgcac gttttgttgg
      421 tagtgatgaa ctggcactgg ttctgggttt tgcaccgggt gatgttgttc tgacaccggc
      481 agttgttctg gcacatctgc atcctgatga tcgtctggaa tggcaggcag gtctgcagcg
      541 ttgtctggca accggtcgtc cggtggttgt taatcatctg ctgctgaccg cagaagcaga
      601 accgcgtccg gcaatgacaa ccctgaccgc actgaccgaa caggatcgtg tgcgtgcagt
      661 taccggtgtt attaccgatc tgagcgatcg tgttcgtcgt gcaaccgaag cagaaattcg
      721 tcaggcagtt cgtgcagcag cagcaacccg tagcgaaatt gatcaggcaa aaggtattgt
      781 tatggcagcc tttgatgttg atgcagatca ggcatttgca ctgctgaaat ggcatagcag
      841 ccagagcaat cgtaaactgc gtgatctggc caccggtatg attgaaggtc tggcagcagc
      901 caatagcgca ctgccgctgc gtcgtcgtct gagcaccgtt tttaccgata tgggttgtcc
      961 ggcaccgagc accaaaggtt ggaccgttcc ggtgaccgat attggtctgc ctccgaccag
     1021 cggtctgatt ccgacagcac tgctgcctgg tattctgacc cgtgcagcac atgatgcaag
     1081 cgttgcaatt accgttgcag atgttaccgc accggatcag cctctggttt atgcaaatcc
     1141 tgcatttgaa cgtctgaccg gttatgcagc agccgaagtt ctgggtcgta attgtcgttt
     1201 tctgcaggca gaaagcggtg atccgcatga acgtagcgca attcgtagcg ccattgcaaa
     1261 tggtgatgca gtgaccacac tgattcgtaa ttttcgtcag gatggtcatg ccttttggaa
     1321 cgaatttcat ctgagtccgg ttcgtaatgg tgcaggtcgt gttacccatt acattggtta
     1381 tcagctggat gtgaccgaac gtgttgaacg tgatcagcag ctggaacagc tggcaagcga
     1441 acagaaactg atttctgaag aagatctgta agatccggct gctaacaaag cccgaaagga
     1501 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1561 acgggtcttg aggggttttt tgctcggccg cactcgagag cttggctgtt ttggcggatg
     1621 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
     1681 gaatttgccg gtgttgacat aaataccact ggcggtgata ctgagcacat cagcaggccc
     1741 ctctagaaat aattttgttt aactttaaat cttcagcagc gaggaggaga tataccatga
     1801 gcacaaaaaa gaaaccatta acacaagagc agcttgagga cgcacgtcgc cttaaagcaa
     1861 tttatgaaaa aaagaaaaat gaacttggct tatcccagga atctgtcgca gacaagatgg
     1921 ggatggggca gtcaggcgtt ggtgctttat ttaatggcat caatgcatta aatgcttata
     1981 acgccgcatt gcttgcaaaa attctcaaag ttagcgttga agaatttagc ccttcaatcg
     2041 ccagagaaat ctacgagatg tatgaagcgg ttagtatgca gccgtcactt agaagtgagt
     2101 atgagtaccc tgttttttct catgttcagg cagggatgtt ctcacctgag cttagaacct
     2161 ttaccaaagg tgatgcggag agatgggtaa gcacaaccaa aaaagccagt gattctgcat
     2221 tctggcttga ggttgaaggt aattccatga ccgcaccaac aggctccaag ccaagctttc
     2281 ctgacggaat gttaattctc gttgaccctg agcaggctgt tgagccaggt gatttctgca
     2341 tagccagact tgggggtgat gagtttacct tcaagaaact gatcagggat agcggtcagg
     2401 tgtttttaca accactaaac ccacagtacc caatgatccc atgcaatgag agttgttccg
     2461 ttgtggggaa agttatcgct agtcagtggc ctgaagagac gtttggcgct gcaaacgacg
     2521 aaaactacgc tttagtagct taataacgct gatagtgcta gtgtagatcg ctactagagc
     2581 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2641 tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg
     2701 cgtttatata ctagagtaac accgtgcgtg ttgactattt tacctctggc ggtgataatg
     2761 gttgctctag aaataatttt gtttaacttt aaatagatca cctgctagcc ggttcagcag
     2821 cgagccgtaa gcctcactgc cggaactgat aggaggagat ataccatggg cagcagccat
     2881 catcatcatc atcacagcag cggcctggtg ccgcgcggca gccatatggg atccgaattc
     2941 gagctccgtc gacaagcttg cggccgcact cgagcatgag atccctggcg gcagtagcgc
     3001 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     3061 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     3121 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     3181 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     3241 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
     3301 gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat
     3361 ccgctcatgg acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc
     3421 ggctatttaa cgaccctgcc ctgaaccgac gaccgggtca tcgtggccgg atcttgcggc
     3481 ccctcggctt gaacgaattg ttagacatta tttgccgact accttggtga tctcgccttt
     3541 cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg
     3601 tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg gcaggcgctc
     3661 cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca
     3721 aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt
     3781 ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa
     3841 gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa
     3901 gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg
     3961 ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc
     4021 gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga
     4081 agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac
     4141 cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta
     4201 caaatgtacg gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga
     4261 tagttgagtc gatacttcgg cgatcaccgc ttccctcata ctcttccttt ttcaatatta
     4321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4381 aaataaacaa atagctagct cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc
     4441 gctgcggaca catacaaagt tacccacaga ttccgtggat aagcagggga ctaacatgtg
     4501 aggcaaaaca gcagggccgc gccggtggcg tttttccata ggctccgccc tcctgccaga
     4561 gttcacataa acagacgctt ttccggtgca tctgtgggag ccgtgaggct caaccatgaa
     4621 tctgacagta cgggcgaaac ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg
     4681 ctccctcttg cgctctcctg ttccgaccct gccgtttacc ggatacctgt tccgcctttc
     4741 tcccttacgg gaagtgtggc gctttctcat agctcacaca ctggtatctc ggctcggtgt
     4801 aggtcgttcg ctccaagctg ggctgtaagc aagaactccc cgttcagccc gactgctgcg
     4861 ccttatccgg taactgttca cttgagtcca acccggaaaa gcacggtaaa acgccactgg
     4921 cagcagccat tggtaactgg gagttcgcag aggatttgtt tagctaaaca cgcggttgct
     4981 cttgaagtgt gcgccaaagt ccggctacac tggaaggaca gatttggttg ctgtgctctg
     5041 cgaaagccag ttaccacggt taagcagttc cccaactgac ttaaccttcg atcaaaccac
     5101 ctccccaggt ggttttttcg tttacagggc aaaagattac gcgcagaaaa aaaggatctc
     5161 aagaagatcc tttgatcttt tctactgaac cgctctagat ttcagtgcaa tttatctctt
     5221 caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcag
     5281 ccccgcatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct
     5341 actccgtcaa gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat
     5401 cattcacttt ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt
     5461 taaatacccg cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga
     5521 taggcatccg ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc
     5581 agcttaagac gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca
     5641 agcaaacatg ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga
     5701 tgtactgaca agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg
     5761 cttccatgcg ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc
     5821 gcccttcccc ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt
     5881 gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt
     5941 catgccagta ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg
     6001 gatgacgacc gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc
     6061 aaacaaattc tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa
     6121 tataaccttt cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa
     6181 tcggcgttaa acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat
     6241 tttgcgcttc agccatactt ttcatactcc cgccattcag agaagaaacc a
//
