LOCUS       sequence-309893-        5453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MlaF(1-246)-(14 aa linker)-GCN4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5453)
  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 5453)
  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: pBEL1967
FEATURES             Location/Qualifiers
     source          1..5453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     201..220
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1119..1175
                     /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(1305..1322)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1475..1561
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1653..1680
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1699..1790
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1791..1859,1860..2651)
                     /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(2009..2028)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2693..3148
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2780..2799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2990..3011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3259..3846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3747..3766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4000..4017
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4183..4205
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4343..4362
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4542..5420)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5447..5453,1..278)
                     /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)"
ORIGIN
        1 caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct
       61 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
      121 gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
      181 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
      241 ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttt gggctagccg
      301 aagggtgaat tatggagcag tctgtggcga atttagtcga tatgcgcgat gtcagtttta
      361 cgcgtggcaa tcgctgcatc ttcgataata tttccctgac cgtgccgcga gggaagatca
      421 cggcgatcat ggggccatcg ggcatcggta aaacgacgct actccgtctg attggcgggc
      481 aaatcgcacc agatcatggt gagatccttt tcgatggtga gaatattccg gcgatgtctc
      541 gttcgcgcct gtatacagtg cgcaaacgga tgagcatgtt atttcagtcc ggggcgttgt
      601 tcactgatat gaacgtattt gacaacgtcg cctatccact gcgcgaacat acccaacttc
      661 ccgcgccatt gttgcatagt acggtgatga tgaagctgga ggccgtgggg ctgcgtggag
      721 cggctaaact aatgccttct gaactttccg gtgggatggc gcggcgtgca gcgctggcgc
      781 gtgcgattgc gctggagccg gatctcatca tgtttgatga accttttgtt gggcaagatc
      841 ccatcaccat gggcgtactg gtgaagctga tttctgagct gaacagcgcg ctgggcgtga
      901 cttgtgtggt ggtttctcac gatgtgccgg aagtgttaag tattgcggat cacgcctgga
      961 tcctggcgga caaaaaaatt gtcgctcatg gcagtgccca ggcgttgcag gcgaatcctg
     1021 atccgcgcgt acgtcagttt ctggacggga gtagtggagg cgggtctggt ggcggctccg
     1081 gcggaggtag tcgtatgaag caacttgagg acaaggttga agagttatta agcaaaaatt
     1141 atcatcttga aaatgaagtc gcgcgtttaa agaagttagt tggcgagcgc taaggtacca
     1201 aattccagaa aagagacgct gaaaagcgtc ttttttcgtt ttggtcctgg tctgaggaat
     1261 ggttgtaagc ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa
     1321 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc
     1381 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg
     1441 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa
     1501 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa
     1561 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg
     1621 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
     1681 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1741 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1801 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1861 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1921 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1981 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     2041 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2101 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2161 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2221 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2281 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     2341 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2401 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2461 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2521 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2581 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2641 agcattggta actgtcagac caagtttact catatatact ttagattgat ttacgcgccc
     2701 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2761 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2821 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2881 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     2941 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3001 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     3061 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3121 tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa
     3181 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3241 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3301 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3361 tccgaaggta actggctcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3421 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3481 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3541 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3601 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3661 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3721 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3781 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3841 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3901 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3961 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4021 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     4081 atagggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4141 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4201 agaggttttc accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc
     4261 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     4321 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     4381 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctgctca tgtttgacag
     4441 cttatcatcg atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta
     4501 tgctactccg tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct
     4561 acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat
     4621 tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg
     4681 gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg
     4741 cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc
     4801 gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg
     4861 ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta
     4921 atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa
     4981 tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc
     5041 tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc
     5101 cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc
     5161 tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt
     5221 cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg
     5281 agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc
     5341 tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga
     5401 tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga aac
//
