LOCUS       sequence-458345-        3427 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pBAD promoter controls the expression of sfGFP. In the presence of 
            VVD-AraC, blue light and L-arabinose can be used to activate sfGFP 
            expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3427)
  AUTHORS   Hollo G, Park JH, Boni E, Schaerli Y
  TITLE     GRN_modeler: An Intuitive Tool for Constructing and Evaluating Gene 
            Regulatory Networks and its Applications to Oscillators and a Light 
            Biosensor
  JOURNAL   bioRxiv 2024.12.18.629005
REFERENCE   2  (bases 1 to 3427)
  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: "bioRxiv 
            2024.12.18.629005"
COMMENT     Sequence Label: pJP_Ctrl04
FEATURES             Location/Qualifiers
     source          1..3427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             121..143
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             151..864
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(955..973)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      969..1016
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1025..1111
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1203..1230
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1418..2233)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1418..2233)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2141..2160
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2234..2325)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2343..2978)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(2488..2507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3133..3417
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3340..3359
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 catttcaccc aaggagctcg ttgagttacc tgcagcgatt gtactagttg cggccgcctg
       61 cgcaccacct gcaggacctc agcaatcgct tggaccagct ttcttaaggg taccaataat
      121 tttgtttaac tttaagaagg agatatacat atgcgtaaag gcgaagaact gtttaccggt
      181 gtggttccga ttctggtgga actggacggc gatgttaatg gtcataaatt cagtgttcgc
      241 ggcgaaggtg aaggcgatgc gacgaacggc aaactgaccc tgaaatttat ctgcaccacg
      301 ggtaaactgc cggtcccgtg gccgacgctg gtgaccacgc tgacctatgg cgttcaatgt
      361 tttgcgcgtt acccggatca catgaaacag cacgactttt tcaaatcggc catgccggaa
      421 ggctatgtgc aggaacgtac gattagcttt aaagacgatg gtacgtataa aacccgcgcg
      481 gaagtgaaat tcgaaggcga taccctggtt aaccgtatcg aactgaaagg tatcgatttc
      541 aaagaagacg gcaatattct gggtcataaa ctggaatata acttcaattc ccacaacgtg
      601 tacatcaccg cggataaaca gaaaaacggc attaaagcca atttcaaaat ccgccataat
      661 gtggaagatg gtagcgttca gctggccgac cactatcagc aaaacacgcc gattggtgat
      721 ggcccggtcc tgctgccgga caatcactac ctgagtaccc agtccgtgct gtcaaaagat
      781 ccgaacgaaa aacgtgacca catggtcctg ctggaatttg tgacggctgc gggtatcacc
      841 cacggcatgg acgaactgta taaaaggcct taataaggat ccggcgccgc tcattcgcta
      901 atcgccacga cgcgtagtcg acccgccaat tcccatgggg cgcgcctgct gccaccgctg
      961 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgccag
     1021 gcatcaaata aaacgaaagg ctcagtcgga agactgggcc tttcgtttta tctgttgttt
     1081 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgtgaa
     1141 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaactaa
     1201 gcagaaggcc atcctgacgg atggcctttt tgcgtttcag atctaccggt aaaccagcaa
     1261 tagacataag cggctattta acgaccctgc cctgaaccga cgacaagctg acgaccgggt
     1321 ctccgcaagt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1381 aatacattca aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca
     1441 aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt
     1501 tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc
     1561 ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa
     1621 taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa
     1681 gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat
     1741 cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc
     1801 ggtcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg
     1861 ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg
     1921 ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct
     1981 tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa
     2041 catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc
     2101 catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc
     2161 catataaatc agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt
     2221 gaatatggct catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggc atgctagcgc
     2341 agaaacgtcc tagaagatgc caggaggata cttagcagag agacaataag gccggagcga
     2401 agccgttttt ccataggctc cgcccccctg acgaacatca cgaaatctga cgctcaaatc
     2461 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct gatggctccc
     2521 tcttgcgctc tcctgttccc gtcctgcggc gtccgtgttg tggtggaggc tttacccaaa
     2581 tcaccacgtc ccgttccgtg tagacagttc gctccaagct gggctgtgtg caagaacccc
     2641 ccgttcagcc cgactgctgc gccttatccg gtaactatca tcttgagtcc aacccggaaa
     2701 gacacgacaa aacgccactg gcagcagcca ttggtaactg agaattagtg gatttagata
     2761 tcgagagtct tgaagtggtg gcctaacaga ggctacactg aaaggacagt atttggtatc
     2821 tgcgctccac taaagccagt taccaggtta agcagttccc caactgactt aaccttcgat
     2881 caaaccgcct ccccaggcgg ttttttcgtt tacagagcag gagattacga cgatcgtaaa
     2941 aggatctcaa gaagatcctt tacggattcc cgacaccatc actctagatt tcagtgcaat
     3001 ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat
     3061 gttagtcatg ccccgcgccc accggaagga gctgactggg ttgctcactt ttcatactcc
     3121 cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct
     3181 tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac
     3241 aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa
     3301 aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
     3361 taagattagc ggttcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc
     3421 ctgaatt
//
