LOCUS       sequence-464739-        8529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of Cys-light LetA(I383C) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8529)
  AUTHORS   Santarossa CC, Li Y, Yousef S, Hasdemir HS, Rodriguez CC, Haase MB, 
            Baek M, Coudray N, Pavek JG, Focke KN, Silverberg AL, Bautista C, 
            Yeh J, Marty MT, Baker D, Tajkhorshid E, Ekiert DC, Bhabha G
  TITLE     LetA defines a structurally distinct transporter family involved in 
            lipid trafficking
  JOURNAL   bioRxiv 2025.03.21.644421
REFERENCE   2  (bases 1 to 8529)
  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: "bioRxiv 
            2025.03.21.644421"
COMMENT     Sequence Label: pBEL2889
FEATURES             Location/Qualifiers
     source          1..8529
                     /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             330..347
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             360..380
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(4397..4414)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4567..4653
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4745..4772
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4791..4882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4883..4951,4952..5743)
                     /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(5101..5120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5785..6240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5872..5891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6082..6103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6351..6938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6839..6858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7092..7109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7275..7297
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7435..7454
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7634..8512)
                     /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 tttttgggct agccgaaggg tgaattatgc atcaccacca tcaccatcag catcaacacg
      361 aaaatctgta ttttcagggg atggctctta acacaccaca aattacgccg acaaaaaaga
      421 taacagtgag ggcaatcggc gaggaactgc cgcgtggtga ttaccaacgt tgcccgcaat
      481 gtgacatgct gtttagcctg cccgagataa attctcatca aagtgcctat tgtccgcgct
      541 gtcaggcaaa aattcgtgac gggcgcgact ggtcgctaac gcgcctggcg gcaatggcct
      601 tcaccatgct gttgcttatg ccgtttgcct ggggcgaacc gctgttgcat atctggctgt
      661 taggcattcg tattgacgcc aacgttatgc aaggcatctg gcaaatgacc aaacagggcg
      721 atgcgataac ggggtcgatg gtctttttct ctgttattgg tgccccgctc attctggtga
      781 cctccatagc ttatttatgg tttggtaacc gactgggaat gaatttacgt ccggtactgc
      841 tgatgcttga gcgacttaaa gagtgggtaa tgctcgatat ctacctggtc ggcattggcg
      901 ttgcttctat aaaggtacag gattatgccc atatccaggc tggtgtcggc ttgttctctt
      961 ttgtggcgtt ggtgatttta acgacggtga cgttgtcaca tcttaatgtc gaagaactgt
     1021 gggagcgatt ttatccgcag cgccccgctt gacgaagggt gaattatgac gcgtagggac
     1081 gagaaacttc gtgtctgcct tgggtgccat tttaccggct atccagatca gcgtggtcgc
     1141 tgcccgcgtt gccatatccc gctacgcctg cgtcgccgtc atagtctgca aaaatcttgg
     1201 gcggcgctgt tagcgtcaat cgttttattg ttacctgcca acctgttgcc tatttctatc
     1261 atttatctga acggaggacg gcaggaagat acaattcttt ccggaattat gtcgctggca
     1321 agtagcaaca ttgcggttgc aggaatcgtg tttatcgcca gtattctggt cccgtttact
     1381 aaagtgatcg tcatgttcac tttactgttg agcattcatt ttaaatctca gcaaggttta
     1441 cgcacacgca ttctgttact gcggatggtg acctggattg gtcgctggtc gatgctcgac
     1501 ctgtttgtca tatctttaac catgtcgctg attaatcgcg atcagtgtct cgcttttact
     1561 atgggaccgg ctgcgtttta tttcggcgca gcggtaattt tgactattct tgctgtggaa
     1621 tggctggaca gccgcttact ttgggatgca catgagtcag gaaacgcccg cttcgacgac
     1681 tgaagcgcag attaaaaata aacgccgtat ctcacctttc tggctgctgc cgttcatcgc
     1741 gctaatgatt gccagttggc tgatttggga cagttatcag gaccggggta ataccgtcac
     1801 catcgacttt atgtcggcgg atggtattgt tccaggccgt acgcctgttc gttatcaggg
     1861 cgttgaagtc ggaacagtgc aggatatcag cctcagcgac gatcttcgta agattgaagt
     1921 caaggtcagc atcaagtccg atatgaaaga tgcgctgcgc gaagagactc agttctggct
     1981 ggtgacgcca aaagcatcgt tggcaggtgt ctccgggctg gacgccctcg tcggtgggaa
     2041 ctatatcggc atgatgccgg gtaaaggtaa agagcaggat cactttgtcg cactcgatac
     2101 ccaaccgaaa tatcggctgg acaatggcga tctgatgatc cacctgcaag cccccgatct
     2161 cggttcgctg aacagcggtt cattggtcta tttccgcaag atcccggtgg gaaaagtcta
     2221 cgactatgcc atcaatccca acaagcaagg cgtggtgatt gatgtcctga tcgagcggcg
     2281 ttttaccgat ctggtgaaaa aaggtagccg tttctggaac gtttccggcg ttgatgccaa
     2341 cgtcagtatc agtggcgcga aggtgaaact ggaaagcctg gcggcactgg ttaacggtgc
     2401 gattgccttc gattcaccag aagagtcgaa acctgccgag gcggaagata cctttggtct
     2461 gtatgaagat ctggcccaca gccagcgtgg cgtaataata aaactggaac tgccgagtgg
     2521 ggccggatta accgccgact cgacgccgtt aatgtatcag gggctggaag tcggacagct
     2581 gactaaactg gatttaaatc ctggtggtaa agtcacaggg gaaatgaccg ttgatcccag
     2641 cgtcgttacc ctgcttcgtg aaaatacccg catcgaatta cgcaacccga aattatccct
     2701 tagcgatgct aatctcagcg ccctgctgac cggcaaaacc ttcgagctgg tgcccggcga
     2761 tggcgagcca cgcaaagagt tcgttgttgt gccaggcgaa aaagcactgc tgcatgaacc
     2821 tgatgttctg acgctgaccc tgaccgcgcc ggaaagttac ggtattgatg cgggtcagcc
     2881 gctcattctt cacggcgtgc aggtaggcca ggttatcgat cgtaaactca ccagcaaagg
     2941 cgtcaccttt accgtcgcca tcgagcctca gcatcgcgaa ctggtaaaag gcgatagcaa
     3001 atttgtcgtc aacagccgtg tcgatgtgaa ggtggggctg gatggcgttg agtttctcgg
     3061 tgccagcgcc tcagaatgga tcaatggcgg gatacgtatt ctgccgggcg ataaaggtga
     3121 gatgaaagcc agctatccac tgtatgccaa tttggaaaaa gcgctggaga acagccttag
     3181 cgatttaccc accacaaccg tgagtttgag tgcagagacg ctgccggatg tgcaggcagg
     3241 atcggtagtg ctgtaccgta aatttgaagt tggtgaagtg attaccgtgc gtccgcgagc
     3301 taacgcgttt gatatcgatc tgcatattaa gccggagtat cgcaaccttc tgaccagcaa
     3361 tagcgtgttc tgggcagaag gcggggcgaa agttcagctg aatggtagtg gtctgaccgt
     3421 acaggcatcc ccgctctcca gagcattaaa gggagccatt agcttcgaca acctcagcgg
     3481 tgccagcgcc agtcagcgta aaggcgataa gcgtattctg tatgcttccg aaacagcggc
     3541 ccgtgcggtt ggcgggcaga ttacgcttca cgctttcgat gccggaaaac tggcggtcgg
     3601 gatgccaatt cgctatctcg gtattgatat cgggcaaatc cagacgctgg atctgattac
     3661 cgcacgcaat gaagtacagg caaaggcggt actttacccg gaatatgtcc agacctttgc
     3721 tcgcggtggt acgcgcttct cagtggtcac accgcaaatt tcggcagctg gcgttgagca
     3781 tcttgatact atcctccagc cgtatatcaa cgtcgaacca ggccggggca atcctcgccg
     3841 cgactttgaa ttacaagagg ccaccattac tgattcgcgt tacctggatg gcttaagcat
     3901 tattgttgaa gcgccggaag ccggttcgtt aggcatcggt acgcctgtgc tgttccgtgg
     3961 tctggaagtc ggtacggtta caggaatgac gctggggaca ttgtcagatc gcgtgatgat
     4021 tgcgatgcgc atcagtaaac gctatcaaca cctggtgcgt aacaattccg tcttctggtt
     4081 ggcatcgggt tacagtctgg actttggtct gacgggcggc gtagtgaaaa ccggcacctt
     4141 taaccagttt atccgtggcg gcatcgcctt cgccacgcct ccgggtacgc cactggcacc
     4201 gaaagcccag gaaggcaaac acttcctgtt gcaggaaagt gaaccgaaag agtggcgtga
     4261 atggggaact gcgcttccca aataaggtac caaattccag aaaagagacg ctgaaaagcg
     4321 tcttttttcg ttttggtcct ggtctgagga atggttgtaa gcttggctgt tttggcggat
     4381 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     4441 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     4501 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     4561 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4621 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     4681 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     4741 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     4801 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4861 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4921 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4981 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5041 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5101 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     5161 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5221 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5281 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5341 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5401 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     5461 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5521 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5581 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5641 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5701 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5761 ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5821 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5881 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5941 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6001 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6061 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     6121 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6181 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     6241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctc agcagagcgc
     6481 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7021 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7081 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     7141 tccttacgca tctgtgcggt atttcacacc gcatagggtc atggctgcgc cccgacaccc
     7201 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     7261 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     7321 cgcgaggcag aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     7381 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     7441 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     7501 tccggcgtag aggatctgct catgtttgac agcttatcat cgatgcataa tgtgcctgtc
     7561 aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct
     7621 gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg
     7681 cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt
     7741 gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa
     7801 gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact
     7861 gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg
     7921 cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc
     7981 gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt
     8041 gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa
     8101 tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc
     8161 ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa
     8221 agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct
     8281 ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt
     8341 tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc
     8401 ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat
     8461 gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca
     8521 tactcccgc
//
