LOCUS       sequence-464813-        5903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of LetA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5903)
  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 5903)
  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: pBEL2214
FEATURES             Location/Qualifiers
     source          1..5903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..51)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      204..290
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      382..409
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        428..519
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(520..588,589..1380)
                     /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(738..757)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1422..1877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1509..1528)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1719..1740
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      1988..2575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2476..2495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2729..2746
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2912..2934
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3072..3091
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3271..4149)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4176..4460
                     /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     4383..4402
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             5788..5808
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
ORIGIN
        1 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
       61 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
      121 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
      181 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      241 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
      301 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
      361 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
      421 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      481 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      541 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      601 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      661 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      721 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
      781 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      841 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
      901 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      961 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1021 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     1081 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1141 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1201 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1261 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1321 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1381 ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct gtagcggcgc
     1441 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     1501 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1561 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1621 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1681 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1741 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     1801 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     1861 attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1921 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1981 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2041 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2101 ctggctcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2161 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2221 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2281 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2341 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2401 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2461 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2521 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2581 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2641 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2701 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2761 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagggtcatg
     2821 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2881 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2941 ccgtcatcac cgaaacgcgc gaggcagaag gagatggcgc ccaacagtcc cccggccacg
     3001 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     3061 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     3121 atgccggcca cgatgcgtcc ggcgtagagg atctgctcat gtttgacagc ttatcatcga
     3181 tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat gctactccgt
     3241 caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta catcattcac
     3301 tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac
     3361 ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat
     3421 ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa
     3481 gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac
     3541 atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg
     3601 acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat
     3661 gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc
     3721 cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc
     3781 atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca
     3841 gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg
     3901 accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa
     3961 ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc
     4021 tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt
     4081 taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc
     4141 ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc catattgcat
     4201 cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac cggtaacccc
     4261 gcttattaaa agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa
     4321 aagtgtctat aatcacggca gaaaagtcca cattgattat ttgcacggcg tcacactttg
     4381 ctatgccata gcatttttat ccataagatt agcggatcct acctgacgct ttttatcgca
     4441 actctctact gtttctccat acccgttttt ttgggtgggc tagccgaagg gtgaattatg
     4501 gctcttaaca caccacaaat tacgccgaca aaaaagataa cagtgagggc aatcggcgag
     4561 gaactgccgc gtggtgatta ccaacgttgc ccgcaatgtg acatgctgtt tagcctgccc
     4621 gagataaatt ctcatcaaag tgcctattgt ccgcgctgtc aggcaaaaat tcgtgacggg
     4681 cgcgactggt cgctaacgcg cctggcggca atggccttca ccatgctgtt gcttatgccg
     4741 tttgcctggg gcgaaccgct gttgcatatc tggctgttag gcattcgtat tgacgccaac
     4801 gttatgcaag gcatctggca aatgaccaaa cagggcgatg cgataacggg gtcgatggtc
     4861 tttttctgcg ttattggtgc cccgctcatt ctggtgacct ccatagctta tttatggttt
     4921 ggtaaccgac tgggaatgaa tttacgtccg gtactgctga tgcttgagcg acttaaagag
     4981 tgggtaatgc tcgatatcta cctggtcggc attggcgttg cttctataaa ggtacaggat
     5041 tatgcccata tccaggctgg tgtcggcttg ttctcttttg tggcgttggt gattttaacg
     5101 acggtgacgt tgtcacatct taatgtcgaa gaactgtggg agcgatttta tccgcagcgc
     5161 cccgctacgc gtagggacga gaaacttcgt gtctgtcttg ggtgccattt taccggctat
     5221 ccagatcagc gtggtcgctg cccgcgttgc catatcccgc tacgcctgcg tcgccgtcat
     5281 agtctgcaaa aatgctgggc ggcgctgtta gcgtcaatcg ttttattgtt acctgccaac
     5341 ctgttgccta tttctatcat ttatctgaac ggaggacggc aggaagatac aattctttcc
     5401 ggaattatgt cgctggcaag tagcaacatt gcggttgcag gaatcgtgtt tatcgccagt
     5461 attctggtac cgtttactaa agtgatcgtc atgttcactt tactgttgag cattcatttt
     5521 aaatgccagc aaggtttacg cacacgcatt ctgttactgc ggatggtgac ctggattggt
     5581 cgctggtcga tgctcgacct gtttgtcata tctttaacca tgtcgctgat taatcgcgat
     5641 cagatcctcg cttttactat gggaccggct gcgttttatt tcggcgcagc ggtaattttg
     5701 actattcttg ctgtggaatg gctggacagc cgcttacttt gggatgcaca tgagtcagga
     5761 aacgcccgct tcgacgacca gcatcaacac catcaccacc atcaccattg aggtaccaaa
     5821 ttccagaaaa gagacgctga aaagcgtctt ttttcgtttt ggtcctggtc tgaggaatgg
     5881 ttgtaagctt ggctgttttg gcg
//
