LOCUS       sequence-464728-        8349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of LetA(Delta-ZnR-N Delta-ZnR-C) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8349)
  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 8349)
  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: pBEL2819
FEATURES             Location/Qualifiers
     source          1..8349
                     /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(4217..4234)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4387..4473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4565..4592
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4611..4702
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4703..4771,4772..5563)
                     /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(4921..4940)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5605..6060
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5692..5711)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5902..5923
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6171..6758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6659..6678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6912..6929
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7095..7117
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7255..7274
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7454..8332)
                     /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 ccgcgtagta gtggaggcgg gtctgacggg
      481 cgcgactggt cgctaacgcg cctggcggca atggccttca ccatgctgtt gcttatgccg
      541 tttgcctggg gcgaaccgct gttgcatatc tggctgttag gcattcgtat tgacgccaac
      601 gttatgcaag gcatctggca aatgaccaaa cagggcgatg cgataacggg gtcgatggtc
      661 tttttctgcg ttattggtgc cccgctcatt ctggtgacct ccatagctta tttatggttt
      721 ggtaaccgac tgggaatgaa tttacgtccg gtactgctga tgcttgagcg acttaaagag
      781 tgggtaatgc tcgatatcta cctggtcggc attggcgttg cttctataaa ggtacaggat
      841 tatgcccata tccaggctgg tgtcggcttg ttctcttttg tggcgttggt gattttaacg
      901 acggtgacgt tgtcacatct taatgtcgaa gaactgtggg agcgatttta tccgcagcgc
      961 agtagtggag gcgggtctgg tggcggctcc cgtcgtcgtc atagtctgca aaaatgctgg
     1021 gcggcgctgt tagcgtcaat cgttttattg ttacctgcca acctgttgcc tatttctatc
     1081 atttatctga acggaggacg gcaggaagat acaattcttt ccggaattat gtcgctggca
     1141 agtagcaaca ttgcggttgc aggaatcgtg tttatcgcca gtattctggt accgtttact
     1201 aaagtgatcg tcatgttcac tttactgttg agcattcatt ttaaatgcca gcaaggttta
     1261 cgcacacgca ttctgttact gcggatggtg acctggattg gtcgctggtc gatgctcgac
     1321 ctgtttgtca tatctttaac catgtcgctg attaatcgcg atcagatcct cgcttttact
     1381 atgggaccgg ctgcgtttta tttcggcgca gcggtaattt tgactattct tgctgtggaa
     1441 tggctggaca gccgcttact ttgggatgca catgagtcag gaaacgcccg cttcgacgac
     1501 tgaagcgcag attaaaaata aacgccgtat ctcacctttc tggctgctgc cgttcatcgc
     1561 gctaatgatt gccagttggc tgatttggga cagttatcag gaccggggta ataccgtcac
     1621 catcgacttt atgtcggcgg atggtattgt tccaggccgt acgcctgttc gttatcaggg
     1681 cgttgaagtc ggaacagtgc aggatatcag cctcagcgac gatcttcgta agattgaagt
     1741 caaggtcagc atcaagtccg atatgaaaga tgcgctgcgc gaagagactc agttctggct
     1801 ggtgacgcca aaagcatcgt tggcaggtgt ctccgggctg gacgccctcg tcggtgggaa
     1861 ctatatcggc atgatgccgg gtaaaggtaa agagcaggat cactttgtcg cactcgatac
     1921 ccaaccgaaa tatcggctgg acaatggcga tctgatgatc cacctgcaag cccccgatct
     1981 cggttcgctg aacagcggtt cattggtcta tttccgcaag atcccggtgg gaaaagtcta
     2041 cgactatgcc atcaatccca acaagcaagg cgtggtgatt gatgtcctga tcgagcggcg
     2101 ttttaccgat ctggtgaaaa aaggtagccg tttctggaac gtttccggcg ttgatgccaa
     2161 cgtcagtatc agtggcgcga aggtgaaact ggaaagcctg gcggcactgg ttaacggtgc
     2221 gattgccttc gattcaccag aagagtcgaa acctgccgag gcggaagata cctttggtct
     2281 gtatgaagat ctggcccaca gccagcgtgg cgtaataata aaactggaac tgccgagtgg
     2341 ggccggatta accgccgact cgacgccgtt aatgtatcag gggctggaag tcggacagct
     2401 gactaaactg gatttaaatc ctggtggtaa agtcacaggg gaaatgaccg ttgatcccag
     2461 cgtcgttacc ctgcttcgtg aaaatacccg catcgaatta cgcaacccga aattatccct
     2521 tagcgatgct aatctcagcg ccctgctgac cggcaaaacc ttcgagctgg tgcccggcga
     2581 tggcgagcca cgcaaagagt tcgttgttgt gccaggcgaa aaagcactgc tgcatgaacc
     2641 tgatgttctg acgctgaccc tgaccgcgcc ggaaagttac ggtattgatg cgggtcagcc
     2701 gctcattctt cacggcgtgc aggtaggcca ggttatcgat cgtaaactca ccagcaaagg
     2761 cgtcaccttt accgtcgcca tcgagcctca gcatcgcgaa ctggtaaaag gcgatagcaa
     2821 atttgtcgtc aacagccgtg tcgatgtgaa ggtggggctg gatggcgttg agtttctcgg
     2881 tgccagcgcc tcagaatgga tcaatggcgg gatacgtatt ctgccgggcg ataaaggtga
     2941 gatgaaagcc agctatccac tgtatgccaa tttggaaaaa gcgctggaga acagccttag
     3001 cgatttaccc accacaaccg tgagtttgag tgcagagacg ctgccggatg tgcaggcagg
     3061 atcggtagtg ctgtaccgta aatttgaagt tggtgaagtg attaccgtgc gtccgcgagc
     3121 taacgcgttt gatatcgatc tgcatattaa gccggagtat cgcaaccttc tgaccagcaa
     3181 tagcgtgttc tgggcagaag gcggggcgaa agttcagctg aatggtagtg gtctgaccgt
     3241 acaggcatcc ccgctctcca gagcattaaa gggagccatt agcttcgaca acctcagcgg
     3301 tgccagcgcc agtcagcgta aaggcgataa gcgtattctg tatgcttccg aaacagcggc
     3361 ccgtgcggtt ggcgggcaga ttacgcttca cgctttcgat gccggaaaac tggcggtcgg
     3421 gatgccaatt cgctatctcg gtattgatat cgggcaaatc cagacgctgg atctgattac
     3481 cgcacgcaat gaagtacagg caaaggcggt actttacccg gaatatgtcc agacctttgc
     3541 tcgcggtggt acgcgcttct cagtggtcac accgcaaatt tcggcagctg gcgttgagca
     3601 tcttgatact atcctccagc cgtatatcaa cgtcgaacca ggccggggca atcctcgccg
     3661 cgactttgaa ttacaagagg ccaccattac tgattcgcgt tacctggatg gcttaagcat
     3721 tattgttgaa gcgccggaag ccggttcgtt aggcatcggt acgcctgtgc tgttccgtgg
     3781 tctggaagtc ggtacggtta caggaatgac gctggggaca ttgtcagatc gcgtgatgat
     3841 tgcgatgcgc atcagtaaac gctatcaaca cctggtgcgt aacaattccg tcttctggtt
     3901 ggcatcgggt tacagtctgg actttggtct gacgggcggc gtagtgaaaa ccggcacctt
     3961 taaccagttt atccgtggcg gcatcgcctt cgccacgcct ccgggtacgc cactggcacc
     4021 gaaagcccag gaaggcaaac acttcctgtt gcaggaaagt gaaccgaaag agtggcgtga
     4081 atggggaact gcgcttccca aataaggtac caaattccag aaaagagacg ctgaaaagcg
     4141 tcttttttcg ttttggtcct ggtctgagga atggttgtaa gcttggctgt tttggcggat
     4201 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     4261 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     4321 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     4381 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4441 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     4501 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     4561 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     4621 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4681 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4741 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4801 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4861 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4921 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     4981 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5041 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5101 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5161 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5221 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     5281 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5341 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5401 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5461 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5521 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5581 ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5641 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5701 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5761 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5821 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5881 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5941 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6001 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     6061 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6121 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6181 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6241 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctc agcagagcgc
     6301 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6361 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6421 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6481 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6541 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6601 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6661 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6721 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6781 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6841 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6901 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     6961 tccttacgca tctgtgcggt atttcacacc gcatagggtc atggctgcgc cccgacaccc
     7021 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     7081 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     7141 cgcgaggcag aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     7201 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     7261 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     7321 tccggcgtag aggatctgct catgtttgac agcttatcat cgatgcataa tgtgcctgtc
     7381 aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct
     7441 gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg
     7501 cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt
     7561 gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa
     7621 gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact
     7681 gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg
     7741 cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc
     7801 gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt
     7861 gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa
     7921 tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc
     7981 ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa
     8041 agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct
     8101 ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt
     8161 tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc
     8221 ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat
     8281 gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca
     8341 tactcccgc
//
