LOCUS       sequence-317548-        7659 bp    DNA     circular SYN 15-APR-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 Apr 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        33..317
                     /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     240..259
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1889..1906
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1919..1939
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3550..3567)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3720..3806
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3898..3925
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3944..4035
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4036..4104,4105..4896)
                     /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(4254..4273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4938..5393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5025..5044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5235..5256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5504..6091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5992..6011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6245..6262
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6428..6450
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6588..6607
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(6787..7659,1..6))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 agccatactt ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag
       61 acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct
      121 tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag
      181 tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta
      241 tgccatagca tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact
      301 ctctactgtt tctccatacc cgtttttttg ggctagccga agggtgaatt atggagcagt
      361 ctgtggcgaa tttagtcgat atgcgcgatg tcagttttac gcgtggcaat cgctgcatct
      421 tcgataatat ttccctgacc gtgccgcgag ggaagatcac ggcgatcatg gggccatcgg
      481 gcatcggtaa aacgacgcta ctccgtctga ttggcgggca aatcgcacca gatcatggtg
      541 agatcctttt cgatggtgag aatattccgg cgatgtctcg ttcgcgcctg tatacagtgc
      601 gcaaacggat gagcatgtta tttcagtccg gggcgttgtt cactgatatg aacgtatttg
      661 acaacgtcgc ctatccactg cgcgaacata cccaacttcc cgcgccattg ttgcatagta
      721 cggtgatgat gaagctggag gccgtggggc tgcgtggagc ggctaaacta atgccttctg
      781 aactttccgg tgggatggcg cggcgtgcag cgctggcgcg tgcgattgcg ctggagccgg
      841 atctcatcat gtttgatgaa ccttttgttg ggcaagatcc catcaccatg ggcgtactgg
      901 tgaagctgat ttctgagctg aacagcgcgc tgggcgtgac ttgtgtggtg gtttctcacg
      961 atgtgccgga agtgttaagt attgcggatc acgcctggat cctggcggac aaaaaaattg
     1021 tcgctcatgg cagtgcccag gcgttgcagg cgaatcctga tccgcgcgta cgtcagtttc
     1081 tggacgggta agccactcat gctgttaaat gcgctggcgt cgctcggaca taaagggatt
     1141 aaaaccctga gaacgttcgg gcgggccggg ttaatgttat tcaatgcgct ggtcggcaaa
     1201 ccggaatttc gcaaacatgc gccgctgctg gtgcgccagc tctataatgt cggcgtcctg
     1261 tcgatgctga ttattgtggt ttctggcgtg ttcatcggaa tggtgttggg gctgcaaggt
     1321 tatctggttc tgaccactta tagtgcggaa accagtctgg gtatgctggt ggcgttatcg
     1381 ctactgcgtg aactggggcc ggtggttgcc gcgttgttgt ttgccgggcg tgctggttcg
     1441 gcgctaaccg cagaaatcgg cctgatgcgc gctacagagc aactctccag tatggagatg
     1501 atggcggtgg atccgctgcg tcgggttatt tctccccgtt tctgggctgg ggttatttca
     1561 ttaccactgt tgacggttat tttcgtcgcc gtggggatct ggggcggatc tctggtcggc
     1621 gtcagttgga aaggcattga tagcgggttc ttctggtcgg caatgcaaaa tgccgtcgac
     1681 tggcgtatgg atctggtcaa ctgtctgatt aagagcgtgg tgttcgccat cacggtgacg
     1741 tggatttcgt tgtttaacgg ctacgacgcc atcccgacgt ctgccgggat tagccgggca
     1801 accactcgca ccgttgtcca ctcgtctctg gctgttctgg ggctggattt tgtgctgacc
     1861 gcattgatgt ttgggaattg agttcatgca tcaccaccat caccatcagc atcaacacga
     1921 aaatctgtat tttcagggga tgcaaacgaa aaaaaatgaa atttgggtgg gtatcttttt
     1981 attagcagca ctgctggcgg cgctgtttgt ttgcctgaag gcggcgaacg tgacgtccat
     2041 acgtactgaa ccgacctaca cgctttatgc gacgttcgat aacattggcg gcctgaaagc
     2101 gcgctctccg gtcagtattg gtggcgttgt tgtgggtcgg gtggcggata ttacgctgga
     2161 cccgaaaacc tatctgccgc gcgtaacgct ggaaattgaa caacgttata accacattcc
     2221 agataccagt tcgctgagca ttcgtacttc cggcctgctg ggggaacaat atctggcatt
     2281 aaacgtcggt tttgaagacc cggaactggg gactgctatc ctgaaggatg gcgatacaat
     2341 tcaggacact aagtctgcga tggtgctgga agatctcatt ggtcagttcc tttacggtag
     2401 taaaggcgat gacaataaga atagtggcga tgcgccagct gctgcgccag gtaataatga
     2461 aaccactgaa cctgtgggta caacgaaata atttcaggag aaccgacgca tgtttaaacg
     2521 tttaatgatg gtcgctttgc tggtgattgc acctctgagt gcggcaaccg cggcagacca
     2581 gaccaatccg tataagctga tggacgaggc ggcgcagaaa acgttcgatc gcctgaagaa
     2641 tgagcaaccg caaattcggg ccaacccgga ttatctgcgt accattgttg atcaggaact
     2701 gctgccatac gtacaggtga aatacgccgg tgcgctggtg ctgggccagt attacaagag
     2761 tgcgacccct gctcaacgtg aagcctactt tgccgctttc cgtgagtacc tgaagcaggc
     2821 ttacggtcag gcgctggcga tgtatcacgg tcaaacctat cagattgcgc cagaacagcc
     2881 gctgggcgat aaaaccattg tgcctattcg cgttaccatt attgacccga atggccgtcc
     2941 gccggtgcgt ctggacttcc agtggcgtaa aaactcccag acgggcaatt ggcaggctta
     3001 cgacatgatt gctgaaggcg tcagtatgat caccaccaaa caaaacgagt ggggaacgct
     3061 gctgcgtacc aaaggtatcg acggcctgac tgcgcaactg aaatcgattt ctcaacagaa
     3121 aatcactctg gaagagaaaa aataatgagc gagtcactga gctggatgca gacgggtgac
     3181 acgctggcgt tatccggaga gctggatcag gacgttttgc taccgctttg ggaaatgcgt
     3241 gaggaagcgg tgaaggggat tacctgcatc gatcttagcc gtgtctcccg cgtggatacg
     3301 gggggactgg cactgctgct ccatcttatc gatctggcga aaaagcaggg caacaatgtg
     3361 acgcttcagg gggtaaacga caaagtgtat accctggcga aattgtataa tttgcctgct
     3421 gatgttctgc ctcgttaagg taccaaattc cagaaaagag acgctgaaaa gcgtcttttt
     3481 tcgttttggt cctggtctga ggaatggttg taagcttggc tgttttggcg gatgagagaa
     3541 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     3601 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     3661 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     3721 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     3781 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     3841 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     3901 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa
     3961 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4021 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4081 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4141 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4201 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4261 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     4321 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4381 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4441 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4501 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4561 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4621 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4681 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4741 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4801 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4861 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4921 atactttaga ttgatttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4981 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     5281 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5341 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttaaaagga
     5401 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5461 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5521 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5581 cggatcaaga gctaccaact ctttttccga aggtaactgg ctcagcagag cgcagatacc
     5641 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5701 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5761 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5821 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5881 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5941 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6001 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6061 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6121 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6181 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6241 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     6301 gcatctgtgc ggtatttcac accgcatagg gtcatggctg cgccccgaca cccgccaaca
     6361 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6421 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     6481 cagaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     6541 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     6601 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     6661 tagaggatct gctcatgttt gacagcttat catcgatgca taatgtgcct gtcaaatgga
     6721 cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt
     6781 taccaattat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
     6841 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
     6901 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
     6961 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
     7021 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
     7081 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
     7141 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
     7201 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
     7261 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
     7321 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
     7381 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
     7441 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
     7501 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
     7561 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
     7621 aaacgagtat cccggcagca ggggatcatt ttgcgcttc
//
