LOCUS       sequence-309837-        7435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MlaFED operon in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7435)
  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 7435)
  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: pBEL1244
FEATURES             Location/Qualifiers
     source          1..7435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..316
                     /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     239..258
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1957..1974
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1987..2007
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3325..3342)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3495..3581
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3673..3700
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3719..3810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3811..3879,3880..4671)
                     /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(4029..4048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4713..5168
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4800..4819)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5010..5031
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5279..5866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5767..5786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6020..6037
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6203..6225
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6363..6382
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(6562..7435,1..5))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga
       61 cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt
      121 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
      181 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
      241 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
      301 tctactgttt ctccataccc gtttttttgg gctagccgaa gggtgaatta tggagcagtc
      361 tgtggcgaat ttagtcgata tgcgcgatgt cagttttacg cgtggcaatc gctgcatctt
      421 cgataatatt tccctgaccg tgccgcgagg gaagatcacg gcgatcatgg ggccatcggg
      481 catcggtaaa acgacgctac tccgtctgat tggcgggcaa atcgcaccag atcatggtga
      541 gatccttttc gatggtgaga atattccggc gatgtctcgt tcgcgcctgt atacagtgcg
      601 caaacggatg agcatgttat ttcagtccgg ggcgttgttc actgatatga acgtatttga
      661 caacgtcgcc tatccactgc gcgaacatac ccaacttccc gcgccattgt tgcatagtac
      721 ggtgatgatg aagctggagg ccgtggggct gcgtggagcg gctaaactaa tgccttctga
      781 actttccggt gggatggcgc ggcgtgcagc gctggcgcgt gcgattgcgc tggagccgga
      841 tctcatcatg tttgatgaac cttttgttgg gcaagatccc atcaccatgg gcgtactggt
      901 gaagctgatt tctgagctga acagcgcgct gggcgtgact tgtgtggtgg tttctcacga
      961 tgtgccggaa gtgttaagta ttgcggatca cgcctggatc ctggcggaca aaaaaattgt
     1021 cgctcatggc agtgcccagg cgttgcaggc gaatcctgat ccgcgcgtac gtcagtttct
     1081 ggacgggata gctgacgggc ctgttccgtt ccgctatcct gccggcgatt atcacgctga
     1141 tcttttacca gggagttaag ccactcatgc tgttaaatgc gctggcgtcg ctcggacata
     1201 aagggattaa aaccctgaga acgttcgggc gggccgggtt aatgttattc aatgcgctgg
     1261 tcggcaaacc ggaatttcgc aaacatgcgc cgctgctggt gcgccagctc tataatgtcg
     1321 gcgtcctgtc gatgctgatt attgtggttt ctggcgtgtt catcggaatg gtgttggggc
     1381 tgcaaggtta tctggttctg accacttata gtgcggaaac cagtctgggt atgctggtgg
     1441 cgttatcgct actgcgtgaa ctggggccgg tggttgccgc gttgttgttt gccgggcgtg
     1501 ctggttcggc gctaaccgca gaaatcggcc tgatgcgcgc tacagagcaa ctctccagta
     1561 tggagatgat ggcggtggat ccgctgcgtc gggttatttc tccccgtttc tgggctgggg
     1621 ttatttcatt accactgttg acggttattt tcgtcgccgt ggggatctgg ggcggatctc
     1681 tggtcggcgt cagttggaaa ggcattgata gcgggttctt ctggtcggca atgcaaaatg
     1741 ccgtcgactg gcgtatggat ctggtcaact gtctgattaa gagcgtggtg ttcgccatca
     1801 cggtgacgtg gatttcgttg tttaacggct acgacgccat cccgacgtct gccgggatta
     1861 gccgggcaac cactcgcacc gttgtccact cgtctctggc tgttctgggg ctggattttg
     1921 tgctgaccgc attgatgttt gggaattgag ttcatgcatc accaccatca ccatcagcat
     1981 caacacgaaa atctgtattt tcaggggatg caaacgaaaa aaaatgaaat ttgggtgggt
     2041 atctttttat tagcagcact gctggcggcg ctgtttgttt gcctgaaggc ggcgaacgtg
     2101 acgtccatac gtactgaacc gacctacacg ctttatgcga cgttcgataa cattggcggc
     2161 ctgaaagcgc gctctccggt cagtattggt ggcgttgttg tgggtcgggt ggcggatatt
     2221 acgctggacc cgaaaaccta tctgccgcgc gtaacgctgg aaattgaaca acgttataac
     2281 cacattccag ataccagttc gctgagcatt cgtacttccg gcctgctggg ggaacaatat
     2341 ctggcattaa acgtcggttt tgaagacccg gaactgggga ctgctatcct gaaggatggc
     2401 gatacaattc aggacactaa gtctgcgatg gtgctggaag atctcattgg tcagttcctt
     2461 tacggtagta aaggcgatga caataagaat agtggcgatg cgccagctgc tgcgccaggt
     2521 aataatgaaa ccactgaacc tgtgggtaca acgaaataat ttcaggagaa ccgacgcatg
     2581 tttaaacgtt taatgatggt cgctttgctg gtgattgcac ctctgagtgc ggcaaccgcg
     2641 gcagaccaga ccaatccgta taagctgatg gacgaggcgg cgcagaaaac gttcgatcgc
     2701 ctgaagaatg agcaaccgca aattcgggcc aacccggatt atctgcgtac cattgttgat
     2761 caggaactgc tgccatacgt acaggtgaaa tacgccggtg cgctggtgct gggccagtat
     2821 tacaagagtg cgacccctgc tcaacgtgaa gcctactttg ccgctttccg tgagtacctg
     2881 aagcaggctt acggtcaggc gctggcgatg tatcacggtc aaacctatca gattgcgcca
     2941 gaacagccgc tgggcgataa aaccattgtg cctattcgcg ttaccattat tgacccgaat
     3001 ggccgtccgc cggtgcgtct ggacttccag tggcgtaaaa actcccagac gggcaattgg
     3061 caggcttacg acatgattgc tgaaggcgtc agtatgatca ccaccaaaca aaacgagtgg
     3121 ggaacgctgc tgcgtaccaa aggtatcgac ggcctgactg cgcaactgaa atcgatttct
     3181 caacagaaaa tcactctgga agagaaaaaa taaggtacca aattccagaa aagagacgct
     3241 gaaaagcgtc ttttttcgtt ttggtcctgg tctgaggaat ggttgtaagc ttggctgttt
     3301 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     3361 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     3421 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     3481 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     3541 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     3601 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     3661 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     3721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3781 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3841 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3901 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3961 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4021 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     4081 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4141 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4201 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4261 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4321 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     4381 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4441 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4501 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4561 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4621 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4681 caagtttact catatatact ttagattgat ttacgcgccc tgtagcggcg cattaagcgc
     4741 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4801 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4861 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4921 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4981 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5041 caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5101 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     5161 tacaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5221 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5281 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5341 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggctcag
     5401 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5461 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5521 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5581 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5641 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5701 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5761 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5821 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5881 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5941 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6001 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     6061 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc
     6121 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     6181 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     6241 ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6301 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6361 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6421 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg atgcataatg
     6481 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     6541 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     6601 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     6661 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     6721 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     6781 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     6841 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     6901 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     6961 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     7021 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     7081 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     7141 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     7201 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     7261 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     7321 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     7381 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttca
//
