LOCUS       sequence-314340-        7773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression and Complementation plasmid for MlaD TM swap with LetB 
            TM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7773)
  AUTHORS   Coudray N, Isom GL, MacRae MR, Saiduddin MN, Bhabha G, Ekiert DC
  TITLE     Structure of bacterial phospholipid transporter MlaFEDB with 
            substrate bound.
  JOURNAL   Elife. 2020 Nov 25;9. pii: 62518. doi: 10.7554/eLife.62518.
  PUBMED    33236984
REFERENCE   2  (bases 1 to 7773)
  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: "2020-11-25"; volume: "9. pii: 62518. doi"; pages: " 
            10.7554/eLife.62518"
COMMENT     Sequence Label: pBEL2138
FEATURES             Location/Qualifiers
     source          1..7773
                     /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             1148..1165
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1178..1198
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3641..3658)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3811..3897
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3989..4016
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4035..4126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4127..4195,4196..4987)
                     /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(4345..4364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5029..5484
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5116..5135)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5326..5347
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5595..6182
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6083..6102
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6336..6353
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6519..6541
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6679..6698
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6878..7756)
                     /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 actcatgcat caccaccatc accatcagca tcaacacgaa aatctgtatt ttcaggggat
     1201 gctgttaaat gcgctggcgt cgctcggaca taaagggatt aaaaccctga gaacgttcgg
     1261 gcgggccggg ttaatgttat tcaatgcgct ggtcggcaaa ccggaatttc gcaaacatgc
     1321 gccgctgctg gtgcgccagc tctataatgt cggcgtcctg tcgatgctga ttattgtggt
     1381 ttctggcgtg ttcatcggaa tggtgttggg gctgcaaggt tatctggttc tgaccactta
     1441 tagtgcggaa accagtctgg gtatgctggt ggcgttatcg ctactgcgtg aactggggcc
     1501 ggtggttgcc gcgttgttgt ttgccgggcg tgctggttcg gcgctaaccg cagaaatcgg
     1561 cctgatgcgc gctacagagc aactctccag tatggagatg atggcggtgg atccgctgcg
     1621 tcgggttatt tctccccgtt tctgggctgg ggttatttca ttaccactgt tgacggttat
     1681 tttcgtcgcc gtggggatct ggggcggatc tctggtcggc gtcagttgga aaggcattga
     1741 tagcgggttc ttctggtcgg caatgcaaaa tgccgtcgac tggcgtatgg atctggtcaa
     1801 ctgtctgatt aagagcgtgg tgttcgccat cacggtgacg tggatttcgt tgtttaacgg
     1861 ctacgacgcc atcccgacgt ctgccgggat tagccgggca accactcgca ccgttgtcca
     1921 ctcgtctctg gctgttctgg ggctggattt tgtgctgacc gcattgatgt ttgggaattg
     1981 agttcatgag tcaggaaacg cccgcttcga cgactgaagc gcagattaaa aataaacgcc
     2041 gtatctcacc tttctggctg ctgccgttca tcgcgctaat gattgccagt tggctgattt
     2101 gggacagtta tcaggcgaac gtgacgtcca tacgtactga accgacctac acgctttatg
     2161 cgacgttcga taacattggc ggcctgaaag cgcgctctcc ggtcagtatt ggtggcgttg
     2221 ttgtgggtcg ggtggcggat attacgctgg acccgaaaac ctatctgccg cgcgtaacgc
     2281 tggaaattga acaacgttat aaccacattc cagataccag ttcgctgagc attcgtactt
     2341 ccggcctgct gggggaacaa tatctggcat taaacgtcgg ttttgaagac ccggaactgg
     2401 ggactgctat cctgaaggat ggcgatacaa ttcaggacac taagtctgcg atggtgctgg
     2461 aagatctcat tggtcagttc ctttacggta gtaaaggcga tgacaataag aatagtggcg
     2521 atgcgccagc tgctgcgcca ggtaataatg aaaccactga acctgtgggt acaacgaaat
     2581 aatttcagga gaaccgacgc atgtttaaac gtttaatgat ggtcgctttg ctggtgattg
     2641 cacctctgag tgcggcaacc gcggcagacc agaccaatcc gtataagctg atggacgagg
     2701 cggcgcagaa aacgttcgat cgcctgaaga atgagcaacc gcaaattcgg gccaacccgg
     2761 attatctgcg taccattgtt gatcaggaac tgctgccata cgtacaggtg aaatacgccg
     2821 gtgcgctggt gctgggccag tattacaaga gtgcgacccc tgctcaacgt gaagcctact
     2881 ttgccgcttt ccgtgagtac ctgaagcagg cttacggtca ggcgctggcg atgtatcacg
     2941 gtcaaaccta tcagattgcg ccagaacagc cgctgggcga taaaaccatt gtgcctattc
     3001 gcgttaccat tattgacccg aatggccgtc cgccggtgcg tctggacttc cagtggcgta
     3061 aaaactccca gacgggcaat tggcaggctt acgacatgat tgctgaaggc gtcagtatga
     3121 tcaccaccaa acaaaacgag tggggaacgc tgctgcgtac caaaggtatc gacggcctga
     3181 ctgcgcaact gaaatcgatt tctcaacaga aaatcactct ggaagagaaa aaataatgag
     3241 cgagtcactg agctggatgc agacgggtga cacgctggcg ttatccggag agctggatca
     3301 ggacgttttg ctaccgcttt gggaaatgcg tgaggaagcg gtgaagggga ttacctgcat
     3361 cgatcttagc cgtgtctccc gcgtggatac ggggggactg gcactgctgc tccatcttat
     3421 cgatctggcg aaaaagcagg gcaacaatgt gacgcttcag ggggtaaacg acaaagtgta
     3481 taccctggcg aaattgtata atttgcctgc tgatgttctg cctcgttaag gtaccaaatt
     3541 ccagaaaaga gacgctgaaa agcgtctttt ttcgttttgg tcctggtctg aggaatggtt
     3601 gtaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     3661 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     3721 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     3781 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     3841 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     3901 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     3961 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     4021 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4081 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4141 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4201 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4261 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4321 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     4381 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4441 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4501 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4561 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4621 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     4681 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4741 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4801 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4861 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4921 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4981 ttggtaactg tcagaccaag tttactcata tatactttag attgatttac gcgccctgta
     5041 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5101 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5161 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5221 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5281 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5341 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     5401 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5461 acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc
     5521 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5581 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5641 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5701 aaggtaactg gctcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5761 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5821 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5881 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5941 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6001 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6061 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6121 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6181 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6241 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6301 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6361 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     6421 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6481 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6541 gttttcaccg tcatcaccga aacgcgcgag gcagaaggag atggcgccca acagtccccc
     6601 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     6661 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     6721 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     6781 tcatcgatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct
     6841 actccgtcaa gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat
     6901 cattcacttt ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt
     6961 taaatacccg cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga
     7021 taggcatccg ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc
     7081 agcttaagac gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca
     7141 agcaaacatg ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga
     7201 tgtactgaca agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg
     7261 cttccatgcg ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc
     7321 gcccttcccc ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt
     7381 gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt
     7441 catgccagta ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg
     7501 gatgacgacc gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc
     7561 aaacaaattc tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa
     7621 tataaccttt cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa
     7681 tcggcgttaa acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat
     7741 tttgcgcttc agccatactt ttcatactcc cgc
//
