LOCUS       sequence-464786-        8605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of split-LetA with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8605)
  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 8605)
  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: pBEL2865
FEATURES             Location/Qualifiers
     source          1..8605
                     /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"
     CDS             1688..1711
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4473..4490)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4643..4729
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4821..4848
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4867..4958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4959..5027,5028..5819)
                     /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(5177..5196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5861..6316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5948..5967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6158..6179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6427..7014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6915..6934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7168..7185
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7351..7373
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7511..7530
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7710..8588)
                     /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 gcgccccgct tgacgaaggg tgaattatga cgcgtaggga
     1081 cgagaaactt cgtgtctgcc ttgggtgcca ttttaccggc tatccagatc agcgtggtcg
     1141 ctgcccgcgt tgccatatcc cgctacgcct gcgtcgccgt catagtctgc aaaaatgctg
     1201 ggcggcgctg ttagcgtcaa tcgttttatt gttacctgcc aacctgttgc ctatttctat
     1261 catttatctg aacggaggac ggcaggaaga tacaattctt tccggaatta tgtcgctggc
     1321 aagtagcaac attgcggttg caggaatcgt gtttatcgcc agtattctgg tcccgtttac
     1381 taaagtgatc gtcatgttca ctttactgtt gagcattcat tttaaatgcc agcaaggttt
     1441 acgcacacgc attctgttac tgcggatggt gacctggatt ggtcgctggt cgatgctcga
     1501 cctgtttgtc atatctttaa ccatgtcgct gattaatcgc gatcagatcc tcgcttttac
     1561 tatgggaccg gctgcgtttt atttcggcgc agcggtaatt ttgactattc ttgctgtgga
     1621 atggctggac agccgcttac tttgggatgc acacgagtca ggaaacgccc gcttcgacga
     1681 cgctagttgg agccacccgc agttcgaaaa atgacgaagg gtgaattatg agtcaggaaa
     1741 cgcccgcttc gacgactgaa gcgcagatta aaaataaacg ccgtatctca cctttctggc
     1801 tgctgccgtt catcgcgcta atgattgcca gttggctgat ttgggacagt tatcaggacc
     1861 ggggtaatac cgtcaccatc gactttatgt cggcggatgg tattgttcca ggccgtacgc
     1921 ctgttcgtta tcagggcgtt gaagtcggaa cagtgcagga tatcagcctc agcgacgatc
     1981 ttcgtaagat tgaagtcaag gtcagcatca agtccgatat gaaagatgcg ctgcgcgaag
     2041 agactcagtt ctggctggtg acgccaaaag catcgttggc aggtgtctcc gggctggacg
     2101 ccctcgtcgg tgggaactat atcggcatga tgccgggtaa aggtaaagag caggatcact
     2161 ttgtcgcact cgatacccaa ccgaaatatc ggctggacaa tggcgatctg atgatccacc
     2221 tgcaagcccc cgatctcggt tcgctgaaca gcggttcatt ggtctatttc cgcaagatcc
     2281 cggtgggaaa agtctacgac tatgccatca atcccaacaa gcaaggcgtg gtgattgatg
     2341 tcctgatcga gcggcgtttt accgatctgg tgaaaaaagg tagccgtttc tggaacgttt
     2401 ccggcgttga tgccaacgtc agtatcagtg gcgcgaaggt gaaactggaa agcctggcgg
     2461 cactggttaa cggtgcgatt gccttcgatt caccagaaga gtcgaaacct gccgaggcgg
     2521 aagatacctt tggtctgtat gaagatctgg cccacagcca gcgtggcgta ataataaaac
     2581 tggaactgcc gagtggggcc ggattaaccg ccgactcgac gccgttaatg tatcaggggc
     2641 tggaagtcgg acagctgact aaactggatt taaatcctgg tggtaaagtc acaggggaaa
     2701 tgaccgttga tcccagcgtc gttaccctgc ttcgtgaaaa tacccgcatc gaattacgca
     2761 acccgaaatt atcccttagc gatgctaatc tcagcgccct gctgaccggc aaaaccttcg
     2821 agctggtgcc cggcgatggc gagccacgca aagagttcgt tgttgtgcca ggcgaaaaag
     2881 cactgctgca tgaacctgat gttctgacgc tgaccctgac cgcgccggaa agttacggta
     2941 ttgatgcggg tcagccgctc attcttcacg gcgtgcaggt aggccaggtt atcgatcgta
     3001 aactcaccag caaaggcgtc acctttaccg tcgccatcga gcctcagcat cgcgaactgg
     3061 taaaaggcga tagcaaattt gtcgtcaaca gccgtgtcga tgtgaaggtg gggctggatg
     3121 gcgttgagtt tctcggtgcc agcgcctcag aatggatcaa tggcgggata cgtattctgc
     3181 cgggcgataa aggtgagatg aaagccagct atccactgta tgccaatttg gaaaaagcgc
     3241 tggagaacag ccttagcgat ttacccacca caaccgtgag tttgagtgca gagacgctgc
     3301 cggatgtgca ggcaggatcg gtagtgctgt accgtaaatt tgaagttggt gaagtgatta
     3361 ccgtgcgtcc gcgagctaac gcgtttgata tcgatctgca tattaagccg gagtatcgca
     3421 accttctgac cagcaatagc gtgttctggg cagaaggcgg ggcgaaagtt cagctgaatg
     3481 gtagtggtct gaccgtacag gcatccccgc tctccagagc attaaaggga gccattagct
     3541 tcgacaacct cagcggtgcc agcgccagtc agcgtaaagg cgataagcgt attctgtatg
     3601 cttccgaaac agcggcccgt gcggttggcg ggcagattac gcttcacgct ttcgatgccg
     3661 gaaaactggc ggtcgggatg ccaattcgct atctcggtat tgatatcggg caaatccaga
     3721 cgctggatct gattaccgca cgcaatgaag tacaggcaaa ggcggtactt tacccggaat
     3781 atgtccagac ctttgctcgc ggtggtacgc gcttctcagt ggtcacaccg caaatttcgg
     3841 cagctggcgt tgagcatctt gatactatcc tccagccgta tatcaacgtc gaaccaggcc
     3901 ggggcaatcc tcgccgcgac tttgaattac aagaggccac cattactgat tcgcgttacc
     3961 tggatggctt aagcattatt gttgaagcgc cggaagccgg ttcgttaggc atcggtacgc
     4021 ctgtgctgtt ccgtggtctg gaagtcggta cggttacagg aatgacgctg gggacattgt
     4081 cagatcgcgt gatgattgcg atgcgcatca gtaaacgcta tcaacacctg gtgcgtaaca
     4141 attccgtctt ctggttggca tcgggttaca gtctggactt tggtctgacg ggcggcgtag
     4201 tgaaaaccgg cacctttaac cagtttatcc gtggcggcat cgccttcgcc acgcctccgg
     4261 gtacgccact ggcaccgaaa gcccaggaag gcaaacactt cctgttgcag gaaagtgaac
     4321 cgaaagagtg gcgtgaatgg ggaactgcgc ttcccaaata aggtaccaaa ttccagaaaa
     4381 gagacgctga aaagcgtctt ttttcgtttt ggtcctggtc tgaggaatgg ttgtaagctt
     4441 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     4501 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     4561 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     4621 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     4681 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     4741 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     4801 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     4861 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4921 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4981 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5041 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5101 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5161 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     5221 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5281 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5341 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5401 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5461 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     5521 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5581 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5641 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5701 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5761 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5821 tgtcagacca agtttactca tatatacttt agattgattt acgcgccctg tagcggcgca
     5881 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5941 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6001 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6061 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6121 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6181 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     6241 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6301 ttaacgttta caatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6361 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6421 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6481 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6541 tggctcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     6601 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6661 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6721 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6781 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6841 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6901 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6961 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7021 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7081 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7141 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7201 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat agggtcatgg
     7261 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     7321 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     7381 cgtcatcacc gaaacgcgcg aggcagaagg agatggcgcc caacagtccc ccggccacgg
     7441 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     7501 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     7561 tgccggccac gatgcgtccg gcgtagagga tctgctcatg tttgacagct tatcatcgat
     7621 gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc
     7681 aagccgtcaa ttgtctgatt cgttaccaat tatgacaact tgacggctac atcattcact
     7741 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
     7801 cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
     7861 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
     7921 acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca
     7981 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
     8041 caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg
     8101 cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc
     8161 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
     8221 tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag
     8281 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
     8341 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
     8401 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
     8461 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
     8521 aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct
     8581 tcagccatac ttttcatact cccgc
//
