LOCUS       sequence-464777-        8511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of LetA(D181A) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8511)
  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 8511)
  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: pBEL3014
FEATURES             Location/Qualifiers
     source          1..8511
                     /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             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             361..381
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(4379..4396)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4549..4635
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4727..4754
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4773..4864
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4865..4933,4934..5725)
                     /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(5083..5102)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5767..6222
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5854..5873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6064..6085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6333..6920
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6821..6840
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7074..7091
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7257..7279
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7417..7436
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7616..8494)
                     /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 catcaccacc atcaccatca gcatcaacac
      361 gaaaatctgt attttcaggg gatggctctt aacacaccac aaattacgcc gacaaaaaag
      421 ataacagtga gggcaatcgg cgaggaactg ccgcgtggtg attaccaacg ttgcccgcaa
      481 tgtgacatgc tgtttagcct gcccgagata aattctcatc aaagtgccta ttgtccgcgc
      541 tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc
      601 ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg
      661 ttaggcattc gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc
      721 gatgcgataa cggggtcgat ggtctttttc tgcgttattg gtgccccgct cattctggtg
      781 acctccatag cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg
      841 ctgatgcttg agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc
      901 gttgcttcta taaaggtaca ggcttatgcc catatccagg ctggtgtcgg cttgttctct
      961 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
     1021 tgggagcgat tttatccgca gcgccccgct acgcgtaggg acgagaaact tcgtgtctgt
     1081 cttgggtgcc attttaccgg ctatccagat cagcgtggtc gctgcccgcg ttgccatatc
     1141 ccgctacgcc tgcgtcgccg tcatagtctg caaaaatgct gggcggcgct gttagcgtca
     1201 atcgttttat tgttacctgc caacctgttg cctatttcta tcatttatct gaacggagga
     1261 cggcaggaag atacaattct ttccggaatt atgtcgctgg caagtagcaa cattgcggtt
     1321 gcaggaatcg tgtttatcgc cagtattctg gtaccgttta ctaaagtgat cgtcatgttc
     1381 actttactgt tgagcattca ttttaaatgc cagcaaggtt tacgcacacg cattctgtta
     1441 ctgcggatgg tgacctggat tggtcgctgg tcgatgctcg acctgtttgt catatcttta
     1501 accatgtcgc tgattaatcg cgatcagatc ctcgctttta ctatgggacc ggctgcgttt
     1561 tatttcggcg cagcggtaat tttgactatt cttgctgtgg aatggctgga cagccgctta
     1621 ctttgggatg cacatgagtc aggaaacgcc cgcttcgacg actgaagcgc agattaaaaa
     1681 taaacgccgt atctcacctt tctggctgct gccgttcatc gcgctaatga ttgccagttg
     1741 gctgatttgg gacagttatc aggaccgggg taataccgtc accatcgact ttatgtcggc
     1801 ggatggtatt gttccaggcc gtacgcctgt tcgttatcag ggcgttgaag tcggaacagt
     1861 gcaggatatc agcctcagcg acgatcttcg taagattgaa gtcaaggtca gcatcaagtc
     1921 cgatatgaaa gatgcgctgc gcgaagagac tcagttctgg ctggtgacgc caaaagcatc
     1981 gttggcaggt gtctccgggc tggacgccct cgtcggtggg aactatatcg gcatgatgcc
     2041 gggtaaaggt aaagagcagg atcactttgt cgcactcgat acccaaccga aatatcggct
     2101 ggacaatggc gatctgatga tccacctgca agcccccgat ctcggttcgc tgaacagcgg
     2161 ttcattggtc tatttccgca agatcccggt gggaaaagtc tacgactatg ccatcaatcc
     2221 caacaagcaa ggcgtggtga ttgatgtcct gatcgagcgg cgttttaccg atctggtgaa
     2281 aaaaggtagc cgtttctgga acgtttccgg cgttgatgcc aacgtcagta tcagtggcgc
     2341 gaaggtgaaa ctggaaagcc tggcggcact ggttaacggt gcgattgcct tcgattcacc
     2401 agaagagtcg aaacctgccg aggcggaaga tacctttggt ctgtatgaag atctggccca
     2461 cagccagcgt ggcgtaataa taaaactgga actgccgagt ggggccggat taaccgccga
     2521 ctcgacgccg ttaatgtatc aggggctgga agtcggacag ctgactaaac tggatttaaa
     2581 tcctggtggt aaagtcacag gggaaatgac cgttgatccc agcgtcgtta ccctgcttcg
     2641 tgaaaatacc cgcatcgaat tacgcaaccc gaaattatcc cttagcgatg ctaatctcag
     2701 cgccctgctg accggcaaaa ccttcgagct ggtgcccggc gatggcgagc cacgcaaaga
     2761 gttcgttgtt gtgccaggcg aaaaagcact gctgcatgaa cctgatgttc tgacgctgac
     2821 cctgaccgcg ccggaaagtt acggtattga tgcgggtcag ccgctcattc ttcacggcgt
     2881 gcaggtaggc caggttatcg atcgtaaact caccagcaaa ggcgtcacct ttaccgtcgc
     2941 catcgagcct cagcatcgcg aactggtaaa aggcgatagc aaatttgtcg tcaacagccg
     3001 tgtcgatgtg aaggtggggc tggatggcgt tgagtttctc ggtgccagcg cctcagaatg
     3061 gatcaatggc gggatacgta ttctgccggg cgataaaggt gagatgaaag ccagctatcc
     3121 actgtatgcc aatttggaaa aagcgctgga gaacagcctt agcgatttac ccaccacaac
     3181 cgtgagtttg agtgcagaga cgctgccgga tgtgcaggca ggatcggtag tgctgtaccg
     3241 taaatttgaa gttggtgaag tgattaccgt gcgtccgcga gctaacgcgt ttgatatcga
     3301 tctgcatatt aagccggagt atcgcaacct tctgaccagc aatagcgtgt tctgggcaga
     3361 aggcggggcg aaagttcagc tgaatggtag tggtctgacc gtacaggcat ccccgctctc
     3421 cagagcatta aagggagcca ttagcttcga caacctcagc ggtgccagcg ccagtcagcg
     3481 taaaggcgat aagcgtattc tgtatgcttc cgaaacagcg gcccgtgcgg ttggcgggca
     3541 gattacgctt cacgctttcg atgccggaaa actggcggtc gggatgccaa ttcgctatct
     3601 cggtattgat atcgggcaaa tccagacgct ggatctgatt accgcacgca atgaagtaca
     3661 ggcaaaggcg gtactttacc cggaatatgt ccagaccttt gctcgcggtg gtacgcgctt
     3721 ctcagtggtc acaccgcaaa tttcggcagc tggcgttgag catcttgata ctatcctcca
     3781 gccgtatatc aacgtcgaac caggccgggg caatcctcgc cgcgactttg aattacaaga
     3841 ggccaccatt actgattcgc gttacctgga tggcttaagc attattgttg aagcgccgga
     3901 agccggttcg ttaggcatcg gtacgcctgt gctgttccgt ggtctggaag tcggtacggt
     3961 tacaggaatg acgctgggga cattgtcaga tcgcgtgatg attgcgatgc gcatcagtaa
     4021 acgctatcaa cacctggtgc gtaacaattc cgtcttctgg ttggcatcgg gttacagtct
     4081 ggactttggt ctgacgggcg gcgtagtgaa aaccggcacc tttaaccagt ttatccgtgg
     4141 cggcatcgcc ttcgccacgc ctccgggtac gccactggca ccgaaagccc aggaaggcaa
     4201 acacttcctg ttgcaggaaa gtgaaccgaa agagtggcgt gaatggggaa ctgcgcttcc
     4261 caaataaggt accaaattcc agaaaagaga cgctgaaaag cgtctttttt cgttttggtc
     4321 ctggtctgag gaatggttgt aagcttggct gttttggcgg atgagagaag attttcagcc
     4381 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     4441 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     4501 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     4561 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     4621 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     4681 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     4741 acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa
     4801 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4861 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4921 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4981 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5041 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5101 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5161 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5221 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5281 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5341 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5401 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5461 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5521 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5581 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5641 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5701 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5761 tgatttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5821 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5881 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5941 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     6001 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6061 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     6121 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6181 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag
     6241 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6301 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6361 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6421 ctaccaactc tttttccgaa ggtaactggc tcagcagagc gcagatacca aatactgtcc
     6481 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6541 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6601 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6661 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6721 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6781 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6841 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6901 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6961 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     7021 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     7081 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     7141 gtatttcaca ccgcataggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     7201 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     7261 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agaaggagat
     7321 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     7381 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     7441 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg
     7501 ctcatgtttg acagcttatc atcgatgcat aatgtgcctg tcaaatggac gaagcaggga
     7561 ttctgcaaac cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg
     7621 acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc
     7681 tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat
     7741 tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga
     7801 tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg
     7861 acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt
     7921 ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat
     7981 ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg
     8041 ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt
     8101 cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg
     8161 acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat
     8221 accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca
     8281 aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc
     8341 gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga
     8401 gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc
     8461 ccggcagcag gggatcattt tgcgcttcag ccatactttt catactcccg c
//
