LOCUS       sequence-309843-        5381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MlaF, tagless, araBAD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5381)
  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 5381)
  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: pBEL1266
FEATURES             Location/Qualifiers
     source          1..5381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..310
                     /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     233..252
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(1265..1282)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1435..1521
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1613..1640
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1659..1750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1751..1819,1820..2611)
                     /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(1969..1988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2653..3108
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2740..2759)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2950..2971
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3219..3806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3707..3726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3960..3977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4143..4165
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4303..4322
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4502..5380)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cttttcatac tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc
       61 cgtcactgcg tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa
      121 agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat
      181 aatcacggca gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata
      241 gcatttttat ccataagatt agcggatcct acctgacgct ttttatcgca actctctact
      301 gtttctccat acccgttttt ttgggctagc cgaagggtga attatggagc agtctgtggc
      361 gaatttagtc gatatgcgcg atgtcagttt tacgcgtggc aatcgctgca tcttcgataa
      421 tatttccctg accgtgccgc gagggaagat cacggcgatc atggggccat cgggcatcgg
      481 taaaacgacg ctactccgtc tgattggcgg gcaaatcgca ccagatcatg gtgagatcct
      541 tttcgatggt gagaatattc cggcgatgtc tcgttcgcgc ctgtatacag tgcgcaaacg
      601 gatgagcatg ttatttcagt ccggggcgtt gttcactgat atgaacgtat ttgacaacgt
      661 cgcctatcca ctgcgcgaac atacccaact tcccgcgcca ttgttgcata gtacggtgat
      721 gatgaagctg gaggccgtgg ggctgcgtgg agcggctaaa ctaatgcctt ctgaactttc
      781 cggtgggatg gcgcggcgtg cagcgctggc gcgtgcgatt gcgctggagc cggatctcat
      841 catgtttgat gaaccttttg ttgggcaaga tcccatcacc atgggcgtac tggtgaagct
      901 gatttctgag ctgaacagcg cgctgggcgt gacttgtgtg gtggtttctc acgatgtgcc
      961 ggaagtgtta agtattgcgg atcacgcctg gatcctggcg gacaaaaaaa ttgtcgctca
     1021 tggcagtgcc caggcgttgc aggcgaatcc tgatccgcgc gtacgtcagt ttctggacgg
     1081 gatagctgac gggcctgttc cgttccgcta tcctgccggc gattatcacg ctgatctttt
     1141 accagggagt taaggtacca aattccagaa aagagacgct gaaaagcgtc ttttttcgtt
     1201 ttggtcctgg tctgaggaat ggttgtaagc ttggctgttt tggcggatga gagaagattt
     1261 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     1321 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     1381 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     1441 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1501 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     1561 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     1621 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
     1681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1741 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1801 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1861 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1921 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1981 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2041 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2101 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2161 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2221 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2281 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2341 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2401 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2461 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2521 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2581 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2641 ttagattgat ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2701 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2761 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2821 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2881 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2941 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     3001 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3061 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttaa aaggatctag
     3121 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3181 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3241 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3301 caagagctac caactctttt tccgaaggta actggctcag cagagcgcag ataccaaata
     3361 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3421 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3481 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3541 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3601 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3661 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3721 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3781 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3841 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3901 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3961 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4021 tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc cgacacccgc caacacccgc
     4081 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4141 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagaa
     4201 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     4261 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     4321 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     4381 gatctgctca tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag
     4441 cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca
     4501 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
     4561 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
     4621 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
     4681 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
     4741 gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat
     4801 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
     4861 gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat
     4921 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
     4981 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
     5041 atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
     5101 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
     5161 acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
     5221 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
     5281 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
     5341 agtatcccgg cagcagggga tcattttgcg cttcagccat a
//
