LOCUS       sequence-371130-        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  JOURNAL   Unpublished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            "Unpublished"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_GFP_mCherry_++
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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             1312..1320
                     /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             1329..2039
                     /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(1474..1492)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1742..1761
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      2270..2356
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2448..2475
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2494..2585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2586..2654,2655..3446)
                     /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     complement(2804..2823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3488..3943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3575..3594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3785..3806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4054..4642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4543..4562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4796..4813
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4979..5001
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5100..5384
                     /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             5411..5419
                     /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             5428..6141
                     /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
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMTDKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5458..5486)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6081..6101
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      6463..6549
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6641..6668
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     6725..6744
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gtaaggaggt
     1321 tataaaaaat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     1381 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     1441 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     1501 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     1561 tgaagcaccc cgccgacatc cccgactaca tgaagctgtc cttccccgag ggcttcaagt
     1621 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     1681 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     1741 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     1801 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     1861 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     1921 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     1981 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaat
     2041 agagtcgacc tgcaggcatg caagcttggc tgttttggcg gatgagagaa gattttcagc
     2101 ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc
     2161 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     2221 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     2281 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     2341 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     2401 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     2461 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
     2521 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2581 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2641 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2701 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2761 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2821 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2881 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2941 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3001 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3061 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3121 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3181 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3241 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3301 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3361 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3421 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3481 ttgatttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3541 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3601 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3661 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     3721 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3781 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt
     3841 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3901 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttaaaagga tctaggtgaa
     3961 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4021 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4081 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4141 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4201 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4261 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4321 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4381 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4441 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4501 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4561 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4621 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4681 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4741 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4801 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4861 cggtatttca caccgcatag ggtcatggct gcgccccgac acccgccaac acccgctgac
     4921 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4981 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagaaggag
     5041 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagca
     5101 agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt
     5161 ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa
     5221 agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga
     5281 ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga
     5341 tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc
     5401 tagcaggagg taaggaggtt ataaaaaatg agtaaaggag aagaactttt cactggagtt
     5461 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga
     5521 gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga
     5581 aaactacctg ttccatggcc aacacttgtc actactttcg gttatggtgt tcaatgcttt
     5641 gcgagatacc cagatcatat gaaacagcat gactttttca agagtgccat gcccgaaggt
     5701 tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa
     5761 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa
     5821 gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac
     5881 atcatgacag acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt
     5941 gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
     6001 cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc
     6061 aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat
     6121 ggcatggatg aactatacaa ataaaggcgg acataaaaaa aaaaaaaaaa aaaaaaaaaa
     6181 aaaaaatgtc cagacctgca ggcatgcaag ctttaatgcg gtagtttatc actagagtcg
     6241 acctgcaggc atgcaagctt ggctgttttg gcggatgaga gaagattttc agcctgatac
     6301 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg
     6361 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     6421 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     6481 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     6541 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     6601 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
     6661 ggcctttttg cgttttacaa actctgctca tgagcccgaa gtggcgagcc cgatcttccc
     6721 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     6781 ccacgatgcg tccggcgtag aggatctgct catgtttgac agcttatc
//
