LOCUS       sequence-464816-        7232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7232)
  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 7232)
  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: pBEL2782
FEATURES             Location/Qualifiers
     source          1..7232
                     /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             2968..2985
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3100..3117)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3270..3356
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3448..3475
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3494..3585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3586..3654,3655..4446)
                     /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(3804..3823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4488..4943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4575..4594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4785..4806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5054..5641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5542..5561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5795..5812
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5978..6000
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6138..6157
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(6337..7215)
                     /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 agtcaggaaa cgcccgcttc gacgactgaa
      361 gcgcagatta aaaataaacg ccgtatctca cctttctggc tgctgccgtt catcgcgcta
      421 atgattgcca gttggctgat ttgggacagt tatcaggacc ggggtaatac cgtcaccatc
      481 gactttatgt cggcggatgg tattgttcca ggccgtacgc ctgttcgtta tcagggcgtt
      541 gaagtcggaa cagtgcagga tatcagcctc agcgacgatc ttcgtaagat tgaagtcaag
      601 gtcagcatca agtccgatat gaaagatgcg ctgcgcgaag agactcagtt ctggctggtg
      661 acgccaaaag catcgttggc aggtgtctcc gggctggacg ccctcgtcgg tgggaactat
      721 atcggcatga tgccgggtaa aggtaaagag caggatcact ttgtcgcact cgatacccaa
      781 ccgaaatatc ggctggacaa tggcgatctg atgatccacc tgcaagcccc cgatctcggt
      841 tcgctgaaca gcggttcatt ggtctatttc cgcaagatcc cggtgggaaa agtctacgac
      901 tatgccatca atcccaacaa gcaaggcgtg gtgattgatg tcctgatcga gcggcgtttt
      961 accgatctgg tgaaaaaagg tagccgtttc tggaacgttt ccggcgttga tgccaacgtc
     1021 agtatcagtg gcgcgaaggt gaaactggaa agcctggcgg cactggttaa cggtgcgatt
     1081 gccttcgatt caccagaaga gtcgaaacct gccgaggcgg aagatacctt tggtctgtat
     1141 gaagatctgg cccacagcca gcgtggcgta ataataaaac tggaactgcc gagtggggcc
     1201 ggattaaccg ccgactcgac gccgttaatg tatcaggggc tggaagtcgg acagctgact
     1261 aaactggatt taaatcctgg tggtaaagtc acaggggaaa tgaccgttga tcccagcgtc
     1321 gttaccctgc ttcgtgaaaa tacccgcatc gaattacgca acccgaaatt atcccttagc
     1381 gatgctaatc tcagcgccct gctgaccggc aaaaccttcg agctggtgcc cggcgatggc
     1441 gagccacgca aagagttcgt tgttgtgcca ggcgaaaaag cactgctgca tgaacctgat
     1501 gttctgacgc tgaccctgac cgcgccggaa agttacggta ttgatgcggg tcagccgctc
     1561 attcttcacg gcgtgcaggt aggccaggtt atcgatcgta aactcaccag caaaggcgtc
     1621 acctttaccg tcgccatcga gcctcagcat cgcgaactgg taaaaggcga tagcaaattt
     1681 gtcgtcaaca gccgtgtcga tgtgaaggtg gggctggatg gcgttgagtt tctcggtgcc
     1741 agcgcctcag aatggatcaa tggcgggata cgtattctgc cgggcgataa aggtgagatg
     1801 aaagccagct atccactgta tgccaatttg gaaaaagcgc tggagaacag ccttagcgat
     1861 ttacccacca caaccgtgag tttgagtgca gagacgctgc cggatgtgca ggcaggatcg
     1921 gtagtgctgt accgtaaatt tgaagttggt gaagtgatta ccgtgcgtcc gcgagctaac
     1981 gcgtttgata tcgatctgca tattaagccg gagtatcgca accttctgac cagcaatagc
     2041 gtgttctggg cagaaggcgg ggcgaaagtt cagctgaatg gtagtggtct gaccgtacag
     2101 gcatccccgc tctccagagc attaaaggga gccattagct tcgacaacct cagcggtgcc
     2161 agcgccagtc agcgtaaagg cgataagcgt attctgtatg cttccgaaac agcggcccgt
     2221 gcggttggcg ggcagattac gcttcacgct ttcgatgccg gaaaactggc ggtcgggatg
     2281 ccaattcgct atctcggtat tgatatcggg caaatccaga cgctggatct gattaccgca
     2341 cgcaatgaag tacaggcaaa ggcggtactt tacccggaat atgtccagac ctttgctcgc
     2401 ggtggtacgc gcttctcagt ggtcacaccg caaatttcgg cagctggcgt tgagcatctt
     2461 gatactatcc tccagccgta tatcaacgtc gaaccaggcc ggggcaatcc tcgccgcgac
     2521 tttgaattac aagaggccac cattactgat tcgcgttacc tggatggctt aagcattatt
     2581 gttgaagcgc cggaagccgg ttcgttaggc atcggtacgc ctgtgctgtt ccgtggtctg
     2641 gaagtcggta cggttacagg aatgacgctg gggacattgt cagatcgcgt gatgattgcg
     2701 atgcgcatca gtaaacgcta tcaacacctg gtgcgtaaca attccgtctt ctggttggca
     2761 tcgggttaca gtctggactt tggtctgacg ggcggcgtag tgaaaaccgg cacctttaac
     2821 cagtttatcc gtggcggcat cgccttcgcc acgcctccgg gtacgccact ggcaccgaaa
     2881 gcccaggaag gcaaacactt cctgttgcag gaaagtgaac cgaaagagtg gcgtgaatgg
     2941 ggaactgcgc ttcccaaaca gcatcaacac catcaccacc atcattaagg taccaaattc
     3001 cagaaaagag acgctgaaaa gcgtcttttt tcgttttggt cctggtctga ggaatggttg
     3061 taagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga
     3121 acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     3181 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     3241 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     3301 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     3361 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     3421 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     3481 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     3841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     4141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttacg cgccctgtag
     4501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4621 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4681 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     4741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4801 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     4861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4921 caaaatatta acgtttacaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4981 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5041 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5101 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5161 aggtaactgg ctcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5221 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5281 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5341 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5401 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5461 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5521 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5581 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5641 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5701 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5761 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5821 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg
     5881 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5941 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     6001 ttttcaccgt catcaccgaa acgcgcgagg cagaaggaga tggcgcccaa cagtcccccg
     6061 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     6121 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     6181 ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt gacagcttat
     6241 catcgatgca taatgtgcct gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta
     6301 ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga cggctacatc
     6361 attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt
     6421 aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat
     6481 aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca
     6541 gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa
     6601 gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat
     6661 gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc
     6721 ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg
     6781 cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg
     6841 cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc
     6901 atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg
     6961 atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca
     7021 aacaaattct cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat
     7081 ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat
     7141 cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt
     7201 ttgcgcttca gccatacttt tcatactccc gc
//
