LOCUS       sequence-371127-        6828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  arabinose-inducible expression of GFP and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6828)
  AUTHORS   Avraham, Roi
  TITLE     Bacterial fluorescent reporters and tags toolkit
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6828)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBAD_mCherry_GFP_-+
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(362..1075)
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(402..422)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1017..1045
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             1084..1092
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(1119..1403)
                     /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     1443..1462
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(1642..2520)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2547..2831
                     /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)"
     RBS             2858..2866
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2875..3585
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYMKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3020..3038)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3288..3307
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      3816..3902
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3994..4021
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4138..4160)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4326..4343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4497..5085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4577..4596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(5196..5651)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5333..5354)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5545..5564
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(5693..6484,6485..6553))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6316..6335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6554..6645)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(6664..6691)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(join(6783..6828,1..41))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc agttccctac
       61 tctcgcatgg ggagacccca cactaccatc ggcgctacgg cgtttcactt ctgagttcgg
      121 catggggtca ggtgggacca ccgcgctact gccgccaggc aaattctgtt ttatcagacc
      181 gcttctgcgt tctgatttaa tctgtatcag gctgaaaatc ttctctcatc cgccaaaaca
      241 gccaagcttg catgcctgca ggtcgactct agtgataaac taccgcatta aagcttgcat
      301 gcctgcaggt ctggacattt tttttttttt tttttttttt ttttttttat gtccgccttt
      361 atttgtatag ttcatccatg ccatgtgtaa tcccagcagc tgttacaaac tcaagaagga
      421 ccatgtggtc tctcttttcg ttgggatctt tcgaaagggc agattgtgtg gacaggtaat
      481 ggttgtctgg taaaaggaca gggccatcgc caattggagt attttgttga taatggtctg
      541 ctagttgaac gcttccatct tcaatgttgt gtctaatttt gaagttaact ttgattccat
      601 tcttttgttt gtctgccatg atgtatacat tgtgtgagtt atagttgtat tccaatttgt
      661 gtccaagaat gtttccatct tctttaaaat caataccttt taactcgatt ctattaacaa
      721 gggtatcacc ttcaaacttg acttcagcac gtgtcttgta gttcccgtca tctttgaaaa
      781 atatagttct ttcctgtaca taaccttcgg gcatggcact cttgaaaaag tcatgctgtt
      841 tcatatgatc tgggtatctc gcaaagcatt gaacaccata accgaaagta gtgacaagtg
      901 ttggccatgg aacaggtagt tttccagtag tgcaaataaa tttaagggta agttttccgt
      961 atgttgcatc accttcaccc tctccactga cagaaaattt gtgcccatta acatcaccat
     1021 ctaattcaac aagaattggg acaactccag tgaaaagttc ttctccttta ctcatttttt
     1081 ataacctcct tacctcctgc tagcccaaaa aaacgggtat ggagaaacag tagagagttg
     1141 cgataaaaag cgtcaggtag gatccgctaa tcttatggat aaaaatgcta tggcatagca
     1201 aagtgtgacg ccgtgcaaat aatcaatgtg gacttttctg ccgtgattat agacactttt
     1261 gttacgcgtt tttgtcatgg ctttggtccc gctttgttac agaatgcttt taataagcgg
     1321 ggttaccggt ttggttagcg agaagagcca gtaaaagacg cagtgacggc aatgtctgat
     1381 gcaatatgga caattggttt cttgctcatg agcccgaagt ggcgagcccg atcttcccca
     1441 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     1501 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg atgcataatg
     1561 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     1621 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     1681 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     1741 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     1801 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     1861 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     1921 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     1981 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     2041 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     2101 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     2161 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     2221 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     2281 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     2341 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     2401 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     2461 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     2521 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg
     2581 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa
     2641 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta
     2701 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat
     2761 agcattttta tccataagat tagcggatcc tacctgacgc tttttatcgc aactctctac
     2821 tgtttctcca tacccgtttt tttgggctag caggaggtaa ggaggttata aaaaatggtg
     2881 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     2941 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     3001 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     3061 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     3121 gacatccccg actacatgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     3181 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     3241 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     3301 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     3361 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     3421 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     3481 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     3541 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaatagag tcgacctgca
     3601 ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     3661 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     3721 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     3781 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     3841 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     3901 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     3961 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     4021 ttgcgtttct acaaactctg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     4081 gggactgttg ggcgccatct ccttctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     4141 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     4201 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccct
     4261 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     4321 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4381 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4441 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4501 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4561 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4621 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4681 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4741 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4801 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4861 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4921 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4981 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5041 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5101 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5161 agattatcaa aaaggatctt cacctagatc cttttaaatt gtaaacgtta atattttgtt
     5221 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5281 caaaatccct tataaatcaa aagaatagcc cgagataggg ttgagtgttg ttccagtttg
     5341 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5401 tcagggcgat ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg
     5461 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5521 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5581 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5641 acagggcgcg taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5701 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5761 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5821 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5881 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5941 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6001 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6061 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6121 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6181 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6241 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6301 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6361 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6421 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6481 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6541 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6601 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt ttgtagaaac
     6661 gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc agtttatggc
     6721 gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc cgctcccggc
     6781 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaa
//
