LOCUS       sequence-300645-        7659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MlaFEDCB operon with MlaF C-terminus deleted [1-246], 
            His-tag on MlaD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7659)
  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 7659)
  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: pBEL1514
FEATURES             Location/Qualifiers
     source          1..7659
                     /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             1866..1883
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1896..1916
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3527..3544)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3697..3783
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3875..3902
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3921..4012
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4013..4081,4082..4873)
                     /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(4231..4250)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4915..5370
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5002..5021)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5212..5233
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5481..6068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5969..5988
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6222..6239
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6405..6427
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6565..6584
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6764..7642)
                     /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 acgggtaagc cactcatgct
     1081 gttaaatgcg ctggcgtcgc tcggacataa agggattaaa accctgagaa cgttcgggcg
     1141 ggccgggtta atgttattca atgcgctggt cggcaaaccg gaatttcgca aacatgcgcc
     1201 gctgctggtg cgccagctct ataatgtcgg cgtcctgtcg atgctgatta ttgtggtttc
     1261 tggcgtgttc atcggaatgg tgttggggct gcaaggttat ctggttctga ccacttatag
     1321 tgcggaaacc agtctgggta tgctggtggc gttatcgcta ctgcgtgaac tggggccggt
     1381 ggttgccgcg ttgttgtttg ccgggcgtgc tggttcggcg ctaaccgcag aaatcggcct
     1441 gatgcgcgct acagagcaac tctccagtat ggagatgatg gcggtggatc cgctgcgtcg
     1501 ggttatttct ccccgtttct gggctggggt tatttcatta ccactgttga cggttatttt
     1561 cgtcgccgtg gggatctggg gcggatctct ggtcggcgtc agttggaaag gcattgatag
     1621 cgggttcttc tggtcggcaa tgcaaaatgc cgtcgactgg cgtatggatc tggtcaactg
     1681 tctgattaag agcgtggtgt tcgccatcac ggtgacgtgg atttcgttgt ttaacggcta
     1741 cgacgccatc ccgacgtctg ccgggattag ccgggcaacc actcgcaccg ttgtccactc
     1801 gtctctggct gttctggggc tggattttgt gctgaccgca ttgatgtttg ggaattgagt
     1861 tcatgcatca ccaccatcac catcagcatc aacacgaaaa tctgtatttt caggggatgc
     1921 aaacgaaaaa aaatgaaatt tgggtgggta tctttttatt agcagcactg ctggcggcgc
     1981 tgtttgtttg cctgaaggcg gcgaacgtga cgtccatacg tactgaaccg acctacacgc
     2041 tttatgcgac gttcgataac attggcggcc tgaaagcgcg ctctccggtc agtattggtg
     2101 gcgttgttgt gggtcgggtg gcggatatta cgctggaccc gaaaacctat ctgccgcgcg
     2161 taacgctgga aattgaacaa cgttataacc acattccaga taccagttcg ctgagcattc
     2221 gtacttccgg cctgctgggg gaacaatatc tggcattaaa cgtcggtttt gaagacccgg
     2281 aactggggac tgctatcctg aaggatggcg atacaattca ggacactaag tctgcgatgg
     2341 tgctggaaga tctcattggt cagttccttt acggtagtaa aggcgatgac aataagaata
     2401 gtggcgatgc gccagctgct gcgccaggta ataatgaaac cactgaacct gtgggtacaa
     2461 cgaaataatt tcaggagaac cgacgcatgt ttaaacgttt aatgatggtc gctttgctgg
     2521 tgattgcacc tctgagtgcg gcaaccgcgg cagaccagac caatccgtat aagctgatgg
     2581 acgaggcggc gcagaaaacg ttcgatcgcc tgaagaatga gcaaccgcaa attcgggcca
     2641 acccggatta tctgcgtacc attgttgatc aggaactgct gccatacgta caggtgaaat
     2701 acgccggtgc gctggtgctg ggccagtatt acaagagtgc gacccctgct caacgtgaag
     2761 cctactttgc cgctttccgt gagtacctga agcaggctta cggtcaggcg ctggcgatgt
     2821 atcacggtca aacctatcag attgcgccag aacagccgct gggcgataaa accattgtgc
     2881 ctattcgcgt taccattatt gacccgaatg gccgtccgcc ggtgcgtctg gacttccagt
     2941 ggcgtaaaaa ctcccagacg ggcaattggc aggcttacga catgattgct gaaggcgtca
     3001 gtatgatcac caccaaacaa aacgagtggg gaacgctgct gcgtaccaaa ggtatcgacg
     3061 gcctgactgc gcaactgaaa tcgatttctc aacagaaaat cactctggaa gagaaaaaat
     3121 aatgagcgag tcactgagct ggatgcagac gggtgacacg ctggcgttat ccggagagct
     3181 ggatcaggac gttttgctac cgctttggga aatgcgtgag gaagcggtga aggggattac
     3241 ctgcatcgat cttagccgtg tctcccgcgt ggatacgggg ggactggcac tgctgctcca
     3301 tcttatcgat ctggcgaaaa agcagggcaa caatgtgacg cttcaggggg taaacgacaa
     3361 agtgtatacc ctggcgaaat tgtataattt gcctgctgat gttctgcctc gttaaggtac
     3421 caaattccag aaaagagacg ctgaaaagcg tcttttttcg ttttggtcct ggtctgagga
     3481 atggttgtaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
     3541 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
     3601 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
     3661 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     3721 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     3781 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
     3841 cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt
     3901 tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct
     3961 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4021 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4081 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4141 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4201 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     4261 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4321 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4381 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4441 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4501 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc
     4561 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4621 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4681 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4741 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4801 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4861 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc
     4921 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4981 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5041 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5101 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     5161 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5221 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     5281 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5341 attttaacaa aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat
     5401 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5461 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5521 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5581 tttccgaagg taactggctc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5641 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5701 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5761 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5821 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5881 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5941 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6001 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6061 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6121 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6181 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6241 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     6301 gcatagggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     6361 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     6421 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag aaggagatgg cgcccaacag
     6481 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     6541 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     6601 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac
     6661 agcttatcat cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc
     6721 tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg
     6781 ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc
     6841 attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg
     6901 tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct
     6961 cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg
     7021 gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat
     7081 cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt
     7141 taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg
     7201 aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg
     7261 gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa
     7321 gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag
     7381 cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg
     7441 gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat
     7501 tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg
     7561 cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg
     7621 gatcattttg cgcttcagcc atacttttca tactcccgc
//
