LOCUS       sequence-370386-        5311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  arabinose-inducible expression of destabilized GFPmut3 (containing a
            AAV C-terminal tail).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5311)
  AUTHORS   Avraham, Roi
  TITLE     Bacterial fluorescent reporters and tags toolkit
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5311)
  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            "Unpublished"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_dGFPmut3-AAV
FEATURES             Location/Qualifiers
     source          1..5311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(105..983)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1010..1294
                     /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     1217..1236
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1321..1329
                     /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             1338..2051
                     /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"
     CDS             1338..2051
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1368..1396)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1991..2011
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2178..2195)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2348..2434
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2526..2553
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2572..2663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2664..2732,2733..3524)
                     /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(2882..2901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3566..4021
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3653..3672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3863..3884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4132..4720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4621..4640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4874..4891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5057..5079
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5217..5236
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cagcttatca tcgatgcata atgtgcctgt caaatggacg aagcagggat tctgcaaacc
       61 ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga caacttgacg
      121 gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg
      181 cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg
      241 gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc
      301 tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac
      361 ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga
      421 tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg
      481 ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc
      541 gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc
      601 ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta
      661 agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga
      721 gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc
      781 ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga
      841 ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg
      901 gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg
      961 ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat
     1021 tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc
     1081 aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca
     1141 aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac
     1201 ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga
     1261 cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc tagcaggagg
     1321 taaggaggtt ataaaaaatg agtaaaggag aagaactttt cactggagtt gtcccaattc
     1381 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag
     1441 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg
     1501 ttccatggcc aacacttgtc actactttcg gttatggtgt tcaatgcttt gcgagatacc
     1561 cagatcatat gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg
     1621 aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg
     1681 aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa
     1741 acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag
     1801 acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa
     1861 gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     1921 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga
     1981 gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg
     2041 aactatacaa aaggcctgca gcaaacgacg aaaactacgc tgcagcagtt taaagcttta
     2101 atgcggtagt ttatcactag agtcgacctg caggcatgca agcttggctg ttttggcgga
     2161 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     2221 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     2281 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     2341 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2401 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     2461 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     2521 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     2581 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2641 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2701 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2761 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2821 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2881 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     2941 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3001 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3061 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3121 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3181 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3241 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3301 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3361 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3421 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3481 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3541 actcatatat actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3601 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3661 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3721 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3781 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3841 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3901 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3961 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4021 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4261 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4741 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4801 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4861 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     4921 tctccttacg catctgtgcg gtatttcaca ccgcataggg tcatggctgc gccccgacac
     4981 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5041 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5101 cgcgcgaggc agaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     5161 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5221 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5281 cgtccggcgt agaggatctg ctcatgtttg a
//
