LOCUS       sequence-300654-        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        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             1935..1952
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1965..1985
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3303..3320)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3473..3559
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3651..3678
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3697..3788
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3789..3857,3858..4649)
                     /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(4007..4026)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4691..5146
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4778..4797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4988..5009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5257..5844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5745..5764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5998..6015
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6181..6203
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6341..6360
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6540..7418)
                     /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 acgggatagc tgacgggcct
     1081 gttccgttcc gctatcctgc cggcgattat cacgctgatc ttttaccagg gagttaagcc
     1141 actcatgctg ttaaatgcgc tggcgtcgct cggacataaa gggattaaaa ccctgagaac
     1201 gttcgggcgg gccgggttaa tgttattcaa tgcgctggtc ggcaaaccgg aatttcgcaa
     1261 acatgcgccg ctgctggtgc gccagctcta taatgtcggc gtcctgtcga tgctgattat
     1321 tgtggtttct ggcgtgttca tcggaatggt gttggggctg caaggttatc tggttctgac
     1381 cacttatagt gcggaaacca gtctgggtat gctggtggcg ttatcgctac tgcgtgaact
     1441 ggggccggtg gttgccgcgt tgttgtttgc cgggcgtgct ggttcggcgc taaccgcaga
     1501 aatcggcctg atgcgcgcta cagagcaact ctccagtatg gagatgatgg cggtggatcc
     1561 gctgcgtcgg gttatttctc cccgtttctg ggctggggtt atttcattac cactgttgac
     1621 ggttattttc gtcgccgtgg ggatctgggg cggatctctg gtcggcgtca gttggaaagg
     1681 cattgatagc gggttcttct ggtcggcaat gcaaaatgcc gtcgactggc gtatggatct
     1741 ggtcaactgt ctgattaaga gcgtggtgtt cgccatcacg gtgacgtgga tttcgttgtt
     1801 taacggctac gacgccatcc cgacgtctgc cgggattagc cgggcaacca ctcgcaccgt
     1861 tgtccactcg tctctggctg ttctggggct ggattttgtg ctgaccgcat tgatgtttgg
     1921 gaattgagtt catgcatcac caccatcacc atcagcatca acacgaaaat ctgtattttc
     1981 aggggatgca aacgaaaaaa aatgaaattt gggtgggtat ctttttatta gcagcactgc
     2041 tggcggcgct gtttgtttgc ctgaaggcgg cgaacgtgac gtccatacgt actgaaccga
     2101 cctacacgct ttatgcgacg ttcgataaca ttggcggcct gaaagcgcgc tctccggtca
     2161 gtattggtgg cgttgttgtg ggtcgggtgg cggatattac gctggacccg aaaacctatc
     2221 tgccgcgcgt aacgctggaa attgaacaac gttataacca cattccagat accagttcgc
     2281 tgagcattcg tacttccggc ctgctggggg aacaatatct ggcattaaac gtcggttttg
     2341 aagacccgga actggggact gctatcctga aggatggcga tacaattcag gacactaagt
     2401 ctgcgatggt gctggaagat ctcattggtc agttccttta cggtagtaaa ggcgatgaca
     2461 ataagaatag tggcgatgcg ccagctgctg cgccaggtaa taatgaaacc actgaacctg
     2521 tgggtacaac gaaataattt caggagaacc gacgcatgtt taaacgttta atgatggtcg
     2581 ctttgctggt gattgcacct ctgagtgcgg caaccgcggc agaccagacc aatccgtata
     2641 agctgatgga cgaggcggcg cagaaaacgt tcgatcgcct gaagaatgag caaccgcaaa
     2701 ttcgggccaa cccggattat ctgcgtacca ttgttgatca ggaactgctg ccatacgtac
     2761 aggtgaaata cgccggtgcg ctggtgctgg gccagtatta caagagtgcg acccctgctc
     2821 aacgtgaagc ctactttgcc gctttccgtg agtacctgaa gcaggcttac ggtcaggcgc
     2881 tggcgatgta tcacggtcaa acctatcaga ttgcgccaga acagccgctg ggcgataaaa
     2941 ccattgtgcc tattcgcgtt accattattg acccgaatgg ccgtccgccg gtgcgtctgg
     3001 acttccagtg gcgtaaaaac tcccagacgg gcaattggca ggcttacgac atgattgctg
     3061 aaggcgtcag tatgatcacc accaaacaaa acgagtgggg aacgctgctg cgtaccaaag
     3121 gtatcgacgg cctgactgcg caactgaaat cgatttctca acagaaaatc actctggaag
     3181 agaaaaaata aggtaccaaa ttccagaaaa gagacgctga aaagcgtctt ttttcgtttt
     3241 ggtcctggtc tgaggaatgg ttgtaagctt ggctgttttg gcggatgaga gaagattttc
     3301 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc
     3361 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     3421 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     3481 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     3541 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     3601 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     3661 cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt
     3721 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3781 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3841 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3901 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3961 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4021 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4081 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4141 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4201 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4261 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4321 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4381 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4441 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4501 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4561 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4621 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4681 agattgattt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4741 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4801 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4861 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4921 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4981 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc gggctattct
     5041 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     5101 caaaaattta acgcgaattt taacaaaata ttaacgttta caatttaaaa ggatctaggt
     5161 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5221 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5281 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5341 agagctacca actctttttc cgaaggtaac tggctcagca gagcgcagat accaaatact
     5401 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5461 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5521 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5581 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5641 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5701 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5761 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5821 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5881 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5941 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6001 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     6061 tgcggtattt cacaccgcat agggtcatgg ctgcgccccg acacccgcca acacccgctg
     6121 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6181 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagaagg
     6241 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6301 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6361 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     6421 tctgctcatg tttgacagct tatcatcgat gcataatgtg cctgtcaaat ggacgaagca
     6481 gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat
     6541 tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc
     6601 gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca
     6661 acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg
     6721 ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga
     6781 tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg
     6841 ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt
     6901 ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt
     6961 atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac
     7021 aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat
     7081 attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg
     7141 tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac
     7201 agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga
     7261 ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa
     7321 tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag
     7381 tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact cccgc
//
