LOCUS       sequence-309871-        5423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MlaF(1-250)-GCN4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5423)
  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 5423)
  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: pBEL1833
FEATURES             Location/Qualifiers
     source          1..5423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..288
                     /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     211..230
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1099..1155
                     /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(1285..1302)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1455..1541
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1633..1660
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1679..1770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1771..1839,1840..2631)
                     /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(1989..2008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2673..3128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2760..2779)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2970..2991
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3239..3826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3727..3746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3980..3997
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4163..4185
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4323..4342
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4522..5400)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc
       61 tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga
      121 ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca
      181 ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag
      241 cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttt
      301 gggctagccg aagggtgaat tatggagcag tctgtggcga atttagtcga tatgcgcgat
      361 gtcagtttta cgcgtggcaa tcgctgcatc ttcgataata tttccctgac cgtgccgcga
      421 gggaagatca cggcgatcat ggggccatcg ggcatcggta aaacgacgct actccgtctg
      481 attggcgggc aaatcgcacc agatcatggt gagatccttt tcgatggtga gaatattccg
      541 gcgatgtctc gttcgcgcct gtatacagtg cgcaaacgga tgagcatgtt atttcagtcc
      601 ggggcgttgt tcactgatat gaacgtattt gacaacgtcg cctatccact gcgcgaacat
      661 acccaacttc ccgcgccatt gttgcatagt acggtgatga tgaagctgga ggccgtgggg
      721 ctgcgtggag cggctaaact aatgccttct gaactttccg gtgggatggc gcggcgtgca
      781 gcgctggcgc gtgcgattgc gctggagccg gatctcatca tgtttgatga accttttgtt
      841 gggcaagatc ccatcaccat gggcgtactg gtgaagctga tttctgagct gaacagcgcg
      901 ctgggcgtga cttgtgtggt ggtttctcac gatgtgccgg aagtgttaag tattgcggat
      961 cacgcctgga tcctggcgga caaaaaaatt gtcgctcatg gcagtgccca ggcgttgcag
     1021 gcgaatcctg atccgcgcgt acgtcagttt ctggacggga tagctgacgg gcgtatgaag
     1081 caacttgagg acaaggttga agagttatta agcaaaaatt atcatcttga aaatgaagtc
     1141 gcgcgtttaa agaagttagt tggcgagcgc taaggtacca aattccagaa aagagacgct
     1201 gaaaagcgtc ttttttcgtt ttggtcctgg tctgaggaat ggttgtaagc ttggctgttt
     1261 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     1321 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     1381 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     1441 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1501 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     1561 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     1621 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     1681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1741 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1801 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1861 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1921 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1981 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     2041 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2101 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2161 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2221 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2281 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     2341 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2401 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2461 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     2521 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2581 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2641 caagtttact catatatact ttagattgat ttacgcgccc tgtagcggcg cattaagcgc
     2701 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2761 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2821 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2881 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2941 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3001 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3061 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3121 tacaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3181 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3241 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3301 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggctcag
     3361 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3421 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3481 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3541 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3601 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3661 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3721 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3781 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3841 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3901 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     3961 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4021 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc
     4081 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4141 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4201 ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     4261 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     4321 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     4381 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg atgcataatg
     4441 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     4501 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     4561 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     4621 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     4681 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     4741 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     4801 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     4861 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     4921 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     4981 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     5041 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     5101 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     5161 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     5221 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     5281 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     5341 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     5401 acttttcata ctcccgccat tca
//
