LOCUS       sequence-318444-        5429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MlaF(1-246)-(6 aa linker)-GCN4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5429)
  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 5429)
  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: pBEL1965
FEATURES             Location/Qualifiers
     source          1..5429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..307
                     /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     230..249
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1124..1180
                     /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(1310..1327)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1480..1566
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1658..1685
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1704..1795
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1796..1864,1865..2656)
                     /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(2014..2033)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2698..3153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2785..2804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2995..3016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3264..3851
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3752..3771
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4005..4022
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4188..4210
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4348..4367
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4547..5425)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt
       61 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc
      121 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat
      181 cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca
      241 tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt
      301 tctccatacc cgtttttttg ggctagccga agggtgaatt atggagcagt ctgtggcgaa
      361 tttagtcgat atgcgcgatg tcagttttac gcgtggcaat cgctgcatct tcgataatat
      421 ttccctgacc gtgccgcgag ggaagatcac ggcgatcatg gggccatcgg gcatcggtaa
      481 aacgacgcta ctccgtctga ttggcgggca aatcgcacca gatcatggtg agatcctttt
      541 cgatggtgag aatattccgg cgatgtctcg ttcgcgcctg tatacagtgc gcaaacggat
      601 gagcatgtta tttcagtccg gggcgttgtt cactgatatg aacgtatttg acaacgtcgc
      661 ctatccactg cgcgaacata cccaacttcc cgcgccattg ttgcatagta cggtgatgat
      721 gaagctggag gccgtggggc tgcgtggagc ggctaaacta atgccttctg aactttccgg
      781 tgggatggcg cggcgtgcag cgctggcgcg tgcgattgcg ctggagccgg atctcatcat
      841 gtttgatgaa ccttttgttg ggcaagatcc catcaccatg ggcgtactgg tgaagctgat
      901 ttctgagctg aacagcgcgc tgggcgtgac ttgtgtggtg gtttctcacg atgtgccgga
      961 agtgttaagt attgcggatc acgcctggat cctggcggac aaaaaaattg tcgctcatgg
     1021 cagtgcccag gcgttgcagg cgaatcctga tccgcgcgta cgtcagtttc tggacgggag
     1081 tagtggaggc gggtctcgta tgaagcaact tgaggacaag gttgaagagt tattaagcaa
     1141 aaattatcat cttgaaaatg aagtcgcgcg tttaaagaag ttagttggcg agcgctaagg
     1201 taccaaattc cagaaaagag acgctgaaaa gcgtcttttt tcgttttggt cctggtctga
     1261 ggaatggttg taagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     1321 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     1381 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     1441 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     1501 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     1561 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     1621 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     1681 ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc
     1741 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1801 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1861 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1921 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1981 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     2041 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2101 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2161 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2221 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2281 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     2341 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2401 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2461 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2521 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2581 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2641 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttacg
     2701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2881 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     2941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3001 tcttgttcca aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag
     3061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3121 cgaattttaa caaaatatta acgtttacaa tttaaaagga tctaggtgaa gatccttttt
     3181 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3241 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3301 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3361 ctttttccga aggtaactgg ctcagcagag cgcagatacc aaatactgtc cttctagtgt
     3421 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3481 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3541 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3601 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3661 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3721 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3781 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3841 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3901 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3961 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4021 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4081 accgcatagg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     4141 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4201 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagaaggaga tggcgcccaa
     4261 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     4321 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     4381 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt
     4441 gacagcttat catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa
     4501 ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga
     4561 cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg
     4621 tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga
     4681 cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt
     4741 cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg
     4801 acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt
     4861 gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact
     4921 cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct
     4981 ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat
     5041 gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt
     5101 taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc
     5161 gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac
     5221 ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga
     5281 gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt
     5341 tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca
     5401 ggggatcatt ttgcgcttca gccatactt
//
