LOCUS       sequence-464801-        8427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of LetA(Delta-ZnR-N) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8427)
  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 8427)
  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: pBEL2913
FEATURES             Location/Qualifiers
     source          1..8427
                     /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(4295..4312)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4465..4551
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4643..4670
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4689..4780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4781..4849,4850..5641)
                     /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(4999..5018)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5683..6138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5770..5789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5980..6001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6249..6836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6737..6756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6990..7007
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7173..7195
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7333..7352
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7532..8410)
                     /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 cccgctacgc gtagggacga gaaacttcgt gtctgtcttg ggtgccattt taccggctat
     1021 ccagatcagc gtggtcgctg cccgcgttgc catatcccgc tacgcctgcg tcgccgtcat
     1081 agtctgcaaa aatgctgggc ggcgctgtta gcgtcaatcg ttttattgtt acctgccaac
     1141 ctgttgccta tttctatcat ttatctgaac ggaggacggc aggaagatac aattctttcc
     1201 ggaattatgt cgctggcaag tagcaacatt gcggttgcag gaatcgtgtt tatcgccagt
     1261 attctggtac cgtttactaa agtgatcgtc atgttcactt tactgttgag cattcatttt
     1321 aaatgccagc aaggtttacg cacacgcatt ctgttactgc ggatggtgac ctggattggt
     1381 cgctggtcga tgctcgacct gtttgtcata tctttaacca tgtcgctgat taatcgcgat
     1441 cagatcctcg cttttactat gggaccggct gcgttttatt tcggcgcagc ggtaattttg
     1501 actattcttg ctgtggaatg gctggacagc cgcttacttt gggatgcaca tgagtcagga
     1561 aacgcccgct tcgacgactg aagcgcagat taaaaataaa cgccgtatct cacctttctg
     1621 gctgctgccg ttcatcgcgc taatgattgc cagttggctg atttgggaca gttatcagga
     1681 ccggggtaat accgtcacca tcgactttat gtcggcggat ggtattgttc caggccgtac
     1741 gcctgttcgt tatcagggcg ttgaagtcgg aacagtgcag gatatcagcc tcagcgacga
     1801 tcttcgtaag attgaagtca aggtcagcat caagtccgat atgaaagatg cgctgcgcga
     1861 agagactcag ttctggctgg tgacgccaaa agcatcgttg gcaggtgtct ccgggctgga
     1921 cgccctcgtc ggtgggaact atatcggcat gatgccgggt aaaggtaaag agcaggatca
     1981 ctttgtcgca ctcgataccc aaccgaaata tcggctggac aatggcgatc tgatgatcca
     2041 cctgcaagcc cccgatctcg gttcgctgaa cagcggttca ttggtctatt tccgcaagat
     2101 cccggtggga aaagtctacg actatgccat caatcccaac aagcaaggcg tggtgattga
     2161 tgtcctgatc gagcggcgtt ttaccgatct ggtgaaaaaa ggtagccgtt tctggaacgt
     2221 ttccggcgtt gatgccaacg tcagtatcag tggcgcgaag gtgaaactgg aaagcctggc
     2281 ggcactggtt aacggtgcga ttgccttcga ttcaccagaa gagtcgaaac ctgccgaggc
     2341 ggaagatacc tttggtctgt atgaagatct ggcccacagc cagcgtggcg taataataaa
     2401 actggaactg ccgagtgggg ccggattaac cgccgactcg acgccgttaa tgtatcaggg
     2461 gctggaagtc ggacagctga ctaaactgga tttaaatcct ggtggtaaag tcacagggga
     2521 aatgaccgtt gatcccagcg tcgttaccct gcttcgtgaa aatacccgca tcgaattacg
     2581 caacccgaaa ttatccctta gcgatgctaa tctcagcgcc ctgctgaccg gcaaaacctt
     2641 cgagctggtg cccggcgatg gcgagccacg caaagagttc gttgttgtgc caggcgaaaa
     2701 agcactgctg catgaacctg atgttctgac gctgaccctg accgcgccgg aaagttacgg
     2761 tattgatgcg ggtcagccgc tcattcttca cggcgtgcag gtaggccagg ttatcgatcg
     2821 taaactcacc agcaaaggcg tcacctttac cgtcgccatc gagcctcagc atcgcgaact
     2881 ggtaaaaggc gatagcaaat ttgtcgtcaa cagccgtgtc gatgtgaagg tggggctgga
     2941 tggcgttgag tttctcggtg ccagcgcctc agaatggatc aatggcggga tacgtattct
     3001 gccgggcgat aaaggtgaga tgaaagccag ctatccactg tatgccaatt tggaaaaagc
     3061 gctggagaac agccttagcg atttacccac cacaaccgtg agtttgagtg cagagacgct
     3121 gccggatgtg caggcaggat cggtagtgct gtaccgtaaa tttgaagttg gtgaagtgat
     3181 taccgtgcgt ccgcgagcta acgcgtttga tatcgatctg catattaagc cggagtatcg
     3241 caaccttctg accagcaata gcgtgttctg ggcagaaggc ggggcgaaag ttcagctgaa
     3301 tggtagtggt ctgaccgtac aggcatcccc gctctccaga gcattaaagg gagccattag
     3361 cttcgacaac ctcagcggtg ccagcgccag tcagcgtaaa ggcgataagc gtattctgta
     3421 tgcttccgaa acagcggccc gtgcggttgg cgggcagatt acgcttcacg ctttcgatgc
     3481 cggaaaactg gcggtcggga tgccaattcg ctatctcggt attgatatcg ggcaaatcca
     3541 gacgctggat ctgattaccg cacgcaatga agtacaggca aaggcggtac tttacccgga
     3601 atatgtccag acctttgctc gcggtggtac gcgcttctca gtggtcacac cgcaaatttc
     3661 ggcagctggc gttgagcatc ttgatactat cctccagccg tatatcaacg tcgaaccagg
     3721 ccggggcaat cctcgccgcg actttgaatt acaagaggcc accattactg attcgcgtta
     3781 cctggatggc ttaagcatta ttgttgaagc gccggaagcc ggttcgttag gcatcggtac
     3841 gcctgtgctg ttccgtggtc tggaagtcgg tacggttaca ggaatgacgc tggggacatt
     3901 gtcagatcgc gtgatgattg cgatgcgcat cagtaaacgc tatcaacacc tggtgcgtaa
     3961 caattccgtc ttctggttgg catcgggtta cagtctggac tttggtctga cgggcggcgt
     4021 agtgaaaacc ggcaccttta accagtttat ccgtggcggc atcgccttcg ccacgcctcc
     4081 gggtacgcca ctggcaccga aagcccagga aggcaaacac ttcctgttgc aggaaagtga
     4141 accgaaagag tggcgtgaat ggggaactgc gcttcccaaa taaggtacca aattccagaa
     4201 aagagacgct gaaaagcgtc ttttttcgtt ttggtcctgg tctgaggaat ggttgtaagc
     4261 ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     4321 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     4381 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     4441 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     4501 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     4561 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     4621 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     4681 caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4741 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4801 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4861 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4921 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4981 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     5041 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5101 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5161 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5221 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5281 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     5341 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5401 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5461 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5521 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5581 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5641 actgtcagac caagtttact catatatact ttagattgat ttacgcgccc tgtagcggcg
     5701 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5761 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5821 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5881 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5941 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     6001 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     6061 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     6121 tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6181 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6361 actggctcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     6421 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6481 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6541 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6601 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6661 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6721 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6781 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6841 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6901 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6961 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     7021 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtcat
     7081 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     7141 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     7201 accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc ccccggccac
     7261 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     7321 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     7381 gatgccggcc acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg
     7441 atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg
     7501 tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca
     7561 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     7621 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     7681 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     7741 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     7801 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     7861 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     7921 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     7981 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     8041 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     8101 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     8161 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     8221 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     8281 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     8341 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     8401 cttcagccat acttttcata ctcccgc
//
