LOCUS       sequence-300267-        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        10..294
                     /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     217..236
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1123..1179
                     /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(1309..1326)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1479..1565
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1657..1684
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1703..1794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1795..1863,1864..2655)
                     /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(2013..2032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2697..3152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2784..2803)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2994..3015
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3263..3850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3751..3770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4004..4021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4187..4209
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4347..4366
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4546..5424)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
       61 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa
      121 gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag
      181 tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa
      241 gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt
      301 ttttttgggc tagccgaagg gtgaattatg gagcagtctg tggcgaattt agtcgatatg
      361 cgcgatgtca gttttacgcg tggcaatcgc tgcatcttcg ataatatttc cctgaccgtg
      421 ccgcgaggga agatcacggc gatcatgggg ccatcgggca tcggtaaaac gacgctactc
      481 cgtctgattg gcgggcaaat cgcaccagat catggtgaga tccttttcga tggtgagaat
      541 attccggcga tgtctcgttc gcgcctgtat acagtgcgca aacggatgag catgttattt
      601 cagtccgggg cgttgttcac tgatatgaac gtatttgaca acgtcgccta tccactgcgc
      661 gaacataccc aacttcccgc gccattgttg catagtacgg tgatgatgaa gctggaggcc
      721 gtggggctgc gtggagcggc taaactaatg ccttctgaac tttccggtgg gatggcgcgg
      781 cgtgcagcgc tggcgcgtgc gattgcgctg gagccggatc tcatcatgtt tgatgaacct
      841 tttgttgggc aagatcccat caccatgggc gtactggtga agctgatttc tgagctgaac
      901 agcgcgctgg gcgtgacttg tgtggtggtt tctcacgatg tgccggaagt gttaagtatt
      961 gcggatcacg cctggatcct ggcggacaaa aaaattgtcg ctcatggcag tgcccaggcg
     1021 ttgcaggcga atcctgatcc gcgcgtacgt cagtttctgg acgggagtag tggaggcggg
     1081 tctggtggcg gctcccgtat gaagcaactt gaggacaagg ttgaagagtt attaagcaaa
     1141 aattatcatc ttgaaaatga agtcgcgcgt ttaaagaagt tagttggcga gcgctaaggt
     1201 accaaattcc agaaaagaga cgctgaaaag cgtctttttt cgttttggtc ctggtctgag
     1261 gaatggttgt aagcttggct gttttggcgg atgagagaag attttcagcc tgatacagat
     1321 taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca gtagcgcggt
     1381 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     1441 ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     1501 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1561 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     1621 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
     1681 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
     1741 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1921 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1981 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     2041 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2101 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2161 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2221 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2281 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     2341 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2401 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2461 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2521 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2581 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2641 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttacgc
     2701 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2761 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2821 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2881 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     2941 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3001 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     3061 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3121 gaattttaac aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg
     3181 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3241 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3301 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3361 tttttccgaa ggtaactggc tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3421 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3481 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3541 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3601 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3661 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3721 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3781 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3841 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3901 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3961 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4021 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4081 ccgcataggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4141 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4201 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agaaggagat ggcgcccaac
     4261 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
     4321 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
     4381 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg ctcatgtttg
     4441 acagcttatc atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
     4501 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
     4561 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
     4621 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
     4681 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
     4741 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
     4801 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
     4861 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
     4921 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
     4981 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
     5041 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     5101 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     5161 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     5221 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
     5281 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     5341 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     5401 gggatcattt tgcgcttcag ccatactttt catactcccg c
//
