LOCUS       sequence-464781-        8433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of LetA(Delta-ZnR-C) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8433)
  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 8433)
  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: pBEL2914
FEATURES             Location/Qualifiers
     source          1..8433
                     /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(4301..4318)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4471..4557
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4649..4676
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4695..4786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4787..4855,4856..5647)
                     /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(5005..5024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5689..6144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5776..5795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5986..6007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6255..6842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6743..6762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6996..7013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7179..7201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7339..7358
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7538..8416)
                     /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 ggattatgcc catatccagg ctggtgtcgg cttgttctct
      961 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
     1021 tgggagcgat tttatccgca gcgcagtagt ggaggcgggt ctggtggcgg ctcccgtcgc
     1081 cgtcatagtc tgcaaaaatg ctgggcggcg ctgttagcgt caatcgtttt attgttacct
     1141 gccaacctgt tgcctatttc tatcatttat ctgaacggag gacggcagga agatacaatt
     1201 ctttccggaa ttatgtcgct ggcaagtagc aacattgcgg ttgcaggaat cgtgtttatc
     1261 gccagtattc tggtaccgtt tactaaagtg atcgtcatgt tcactttact gttgagcatt
     1321 cattttaaat gccagcaagg tttacgcaca cgcattctgt tactgcggat ggtgacctgg
     1381 attggtcgct ggtcgatgct cgacctgttt gtcatatctt taaccatgtc gctgattaat
     1441 cgcgatcaga tcctcgcttt tactatggga ccggctgcgt tttatttcgg cgcagcggta
     1501 attttgacta ttcttgctgt ggaatggctg gacagccgct tactttggga tgcacatgag
     1561 tcaggaaacg cccgcttcga cgactgaagc gcagattaaa aataaacgcc gtatctcacc
     1621 tttctggctg ctgccgttca tcgcgctaat gattgccagt tggctgattt gggacagtta
     1681 tcaggaccgg ggtaataccg tcaccatcga ctttatgtcg gcggatggta ttgttccagg
     1741 ccgtacgcct gttcgttatc agggcgttga agtcggaaca gtgcaggata tcagcctcag
     1801 cgacgatctt cgtaagattg aagtcaaggt cagcatcaag tccgatatga aagatgcgct
     1861 gcgcgaagag actcagttct ggctggtgac gccaaaagca tcgttggcag gtgtctccgg
     1921 gctggacgcc ctcgtcggtg ggaactatat cggcatgatg ccgggtaaag gtaaagagca
     1981 ggatcacttt gtcgcactcg atacccaacc gaaatatcgg ctggacaatg gcgatctgat
     2041 gatccacctg caagcccccg atctcggttc gctgaacagc ggttcattgg tctatttccg
     2101 caagatcccg gtgggaaaag tctacgacta tgccatcaat cccaacaagc aaggcgtggt
     2161 gattgatgtc ctgatcgagc ggcgttttac cgatctggtg aaaaaaggta gccgtttctg
     2221 gaacgtttcc ggcgttgatg ccaacgtcag tatcagtggc gcgaaggtga aactggaaag
     2281 cctggcggca ctggttaacg gtgcgattgc cttcgattca ccagaagagt cgaaacctgc
     2341 cgaggcggaa gatacctttg gtctgtatga agatctggcc cacagccagc gtggcgtaat
     2401 aataaaactg gaactgccga gtggggccgg attaaccgcc gactcgacgc cgttaatgta
     2461 tcaggggctg gaagtcggac agctgactaa actggattta aatcctggtg gtaaagtcac
     2521 aggggaaatg accgttgatc ccagcgtcgt taccctgctt cgtgaaaata cccgcatcga
     2581 attacgcaac ccgaaattat cccttagcga tgctaatctc agcgccctgc tgaccggcaa
     2641 aaccttcgag ctggtgcccg gcgatggcga gccacgcaaa gagttcgttg ttgtgccagg
     2701 cgaaaaagca ctgctgcatg aacctgatgt tctgacgctg accctgaccg cgccggaaag
     2761 ttacggtatt gatgcgggtc agccgctcat tcttcacggc gtgcaggtag gccaggttat
     2821 cgatcgtaaa ctcaccagca aaggcgtcac ctttaccgtc gccatcgagc ctcagcatcg
     2881 cgaactggta aaaggcgata gcaaatttgt cgtcaacagc cgtgtcgatg tgaaggtggg
     2941 gctggatggc gttgagtttc tcggtgccag cgcctcagaa tggatcaatg gcgggatacg
     3001 tattctgccg ggcgataaag gtgagatgaa agccagctat ccactgtatg ccaatttgga
     3061 aaaagcgctg gagaacagcc ttagcgattt acccaccaca accgtgagtt tgagtgcaga
     3121 gacgctgccg gatgtgcagg caggatcggt agtgctgtac cgtaaatttg aagttggtga
     3181 agtgattacc gtgcgtccgc gagctaacgc gtttgatatc gatctgcata ttaagccgga
     3241 gtatcgcaac cttctgacca gcaatagcgt gttctgggca gaaggcgggg cgaaagttca
     3301 gctgaatggt agtggtctga ccgtacaggc atccccgctc tccagagcat taaagggagc
     3361 cattagcttc gacaacctca gcggtgccag cgccagtcag cgtaaaggcg ataagcgtat
     3421 tctgtatgct tccgaaacag cggcccgtgc ggttggcggg cagattacgc ttcacgcttt
     3481 cgatgccgga aaactggcgg tcgggatgcc aattcgctat ctcggtattg atatcgggca
     3541 aatccagacg ctggatctga ttaccgcacg caatgaagta caggcaaagg cggtacttta
     3601 cccggaatat gtccagacct ttgctcgcgg tggtacgcgc ttctcagtgg tcacaccgca
     3661 aatttcggca gctggcgttg agcatcttga tactatcctc cagccgtata tcaacgtcga
     3721 accaggccgg ggcaatcctc gccgcgactt tgaattacaa gaggccacca ttactgattc
     3781 gcgttacctg gatggcttaa gcattattgt tgaagcgccg gaagccggtt cgttaggcat
     3841 cggtacgcct gtgctgttcc gtggtctgga agtcggtacg gttacaggaa tgacgctggg
     3901 gacattgtca gatcgcgtga tgattgcgat gcgcatcagt aaacgctatc aacacctggt
     3961 gcgtaacaat tccgtcttct ggttggcatc gggttacagt ctggactttg gtctgacggg
     4021 cggcgtagtg aaaaccggca cctttaacca gtttatccgt ggcggcatcg ccttcgccac
     4081 gcctccgggt acgccactgg caccgaaagc ccaggaaggc aaacacttcc tgttgcagga
     4141 aagtgaaccg aaagagtggc gtgaatgggg aactgcgctt cccaaataag gtaccaaatt
     4201 ccagaaaaga gacgctgaaa agcgtctttt ttcgttttgg tcctggtctg aggaatggtt
     4261 gtaagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     4321 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     4381 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     4441 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     4501 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     4561 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     4621 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     4681 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4741 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4801 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4861 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4921 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4981 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     5041 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5101 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5161 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5221 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5281 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     5341 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5401 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5461 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5521 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5581 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5641 ttggtaactg tcagaccaag tttactcata tatactttag attgatttac gcgccctgta
     5701 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5761 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5821 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5881 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5941 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6001 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
     6061 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6121 acaaaatatt aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc
     6181 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6361 aaggtaactg gctcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     6421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7021 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     7081 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     7141 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     7201 gttttcaccg tcatcaccga aacgcgcgag gcagaaggag atggcgccca acagtccccc
     7261 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     7321 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     7381 gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta
     7441 tcatcgatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct
     7501 actccgtcaa gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat
     7561 cattcacttt ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt
     7621 taaatacccg cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga
     7681 taggcatccg ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc
     7741 agcttaagac gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca
     7801 agcaaacatg ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga
     7861 tgtactgaca agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg
     7921 cttccatgcg ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc
     7981 gcccttcccc ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt
     8041 gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt
     8101 catgccagta ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg
     8161 gatgacgacc gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc
     8221 aaacaaattc tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa
     8281 tataaccttt cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa
     8341 tcggcgttaa acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat
     8401 tttgcgcttc agccatactt ttcatactcc cgc
//
