LOCUS       sequence-317549-        5441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MlaF(1-246)-(10 aa linker)-GCN4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5441)
  AUTHORS   Kolich LR, Chang YT, Coudray N, Giacometti SI, MacRae MR, Isom GL, 
            Teran EM, Bhabha G, Ekiert DC
  TITLE     Structure of MlaFB uncovers novel mechanisms of ABC transporter 
            regulation.
  JOURNAL   Elife. 2020 Jun 30;9. pii: 60030. doi: 10.7554/eLife.60030.
  PUBMED    32602838
REFERENCE   2  (bases 1 to 5441)
  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: "Elife."; 
            date: "2020-06-30"; volume: "9. pii: 60030. doi"; pages: " 
            10.7554/eLife.60030"
COMMENT     Sequence Label: pBEL1966
FEATURES             Location/Qualifiers
     source          1..5441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..309
                     /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     232..251
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1138..1194
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     primer_bind     complement(1324..1341)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1494..1580
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1672..1699
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1718..1809
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1810..1878,1879..2670)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2028..2047)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2712..3167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2799..2818)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3009..3030
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3278..3865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3766..3785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4019..4036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4202..4224
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4362..4381
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4561..5439)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc agacattgcc
       61 gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa
      121 gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata
      181 atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag
      241 catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa ctctctactg
      301 tttctccata cccgtttttt tgggctagcc gaagggtgaa ttatggagca gtctgtggcg
      361 aatttagtcg atatgcgcga tgtcagtttt acgcgtggca atcgctgcat cttcgataat
      421 atttccctga ccgtgccgcg agggaagatc acggcgatca tggggccatc gggcatcggt
      481 aaaacgacgc tactccgtct gattggcggg caaatcgcac cagatcatgg tgagatcctt
      541 ttcgatggtg agaatattcc ggcgatgtct cgttcgcgcc tgtatacagt gcgcaaacgg
      601 atgagcatgt tatttcagtc cggggcgttg ttcactgata tgaacgtatt tgacaacgtc
      661 gcctatccac tgcgcgaaca tacccaactt cccgcgccat tgttgcatag tacggtgatg
      721 atgaagctgg aggccgtggg gctgcgtgga gcggctaaac taatgccttc tgaactttcc
      781 ggtgggatgg cgcggcgtgc agcgctggcg cgtgcgattg cgctggagcc ggatctcatc
      841 atgtttgatg aaccttttgt tgggcaagat cccatcacca tgggcgtact ggtgaagctg
      901 atttctgagc tgaacagcgc gctgggcgtg acttgtgtgg tggtttctca cgatgtgccg
      961 gaagtgttaa gtattgcgga tcacgcctgg atcctggcgg acaaaaaaat tgtcgctcat
     1021 ggcagtgccc aggcgttgca ggcgaatcct gatccgcgcg tacgtcagtt tctggacggg
     1081 agtagtggag gcgggtctgg tggcggctcc cgtatgaagc aacttgagga caaggttgaa
     1141 gagttattaa gcaaaaatta tcatcttgaa aatgaagtcg cgcgtttaaa gaagttagtt
     1201 ggcgagcgct aaggtaccaa attccagaaa agagacgctg aaaagcgtct tttttcgttt
     1261 tggtcctggt ctgaggaatg gttgtaagct tggctgtttt ggcggatgag agaagatttt
     1321 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     1381 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     1441 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     1501 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1561 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     1621 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     1681 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     1741 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1801 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     1861 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1921 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1981 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2041 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2101 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2161 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2221 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2281 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2341 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2401 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2461 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2521 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2581 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2641 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2701 tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2761 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2821 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2881 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     2941 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3001 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     3061 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3121 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa aggatctagg
     3181 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3241 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3301 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3361 aagagctacc aactcttttt ccgaaggtaa ctggctcagc agagcgcaga taccaaatac
     3421 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3481 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3541 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3601 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3661 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3721 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3781 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3841 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3901 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3961 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     4021 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     4081 gtgcggtatt tcacaccgca tagggtcatg gctgcgcccc gacacccgcc aacacccgct
     4141 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4201 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagaag
     4261 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     4321 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     4381 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     4441 atctgctcat gtttgacagc ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc
     4501 agggattctg caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa
     4561 ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct
     4621 cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc
     4681 aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg
     4741 gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag
     4801 atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt
     4861 gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg
     4921 tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt
     4981 tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa
     5041 caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa
     5101 tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg
     5161 gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa
     5221 cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg
     5281 accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa
     5341 atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga
     5401 gtatcccggc agcaggggat cattttgcgc ttcagccata c
//
