LOCUS       sequence-464799-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA(C233A) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7073)
  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 7073)
  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: pBEL2421
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..111
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(112..180,181..972)
                     /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(330..349)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1143..1731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1632..1651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1885..1902
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1917..2056
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1982..2001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2142..2164
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2158..2349)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2858..2877
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2858..2876
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2908..2930
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6888..6906)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6902..6949
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(6994..7012)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
       61 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      121 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      181 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      241 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      301 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
      361 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      421 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
      481 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
      541 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      601 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
      661 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      721 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      781 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      841 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
      901 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
      961 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     1021 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1081 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1141 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1201 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1261 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     1321 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1381 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1441 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1501 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1561 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1621 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1681 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1741 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     1801 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     1861 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     1921 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     1981 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     2041 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     2101 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     2161 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     2221 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     2281 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     2341 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     2401 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     2461 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     2521 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     2581 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     2641 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     2701 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     2761 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggcca
     2821 ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac tatagggaga
     2881 ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     2941 gctcttaaca caccacaaat tacgccgaca aaaaagataa cagtgagggc aatcggcgag
     3001 gaactgccgc gtggtgatta ccaacgttgc ccgcaatgtg acatgctgtt tagcctgccc
     3061 gagataaatt ctcatcaaag tgcctattgt ccgcgctgtc aggcaaaaat tcgtgacggg
     3121 cgcgactggt cgctaacgcg cctggcggca atggccttca ccatgctgtt gcttatgccg
     3181 tttgcctggg gcgaaccgct gttgcatatc tggctgttag gcattcgtat tgacgccaac
     3241 gttatgcaag gcatctggca aatgaccaaa cagggcgatg cgataacggg gtcgatggtc
     3301 tttttctgcg ttattggtgc cccgctcatt ctggtgacct ccatagctta tttatggttt
     3361 ggtaaccgac tgggaatgaa tttacgtccg gtactgctga tgcttgagcg acttaaagag
     3421 tgggtaatgc tcgatatcta cctggtcggc attggcgttg cttctataaa ggtacaggat
     3481 tatgcccata tccaggctgg tgtcggcttg ttctcttttg tggcgttggt gattttaacg
     3541 acggtgacgt tgtcacatct taatgtcgaa gaactgtggg agcgatttta tccgcagcgc
     3601 cccgctacgc gtagggacga gaaacttcgt gtcgctcttg ggtgccattt taccggctat
     3661 ccagatcagc gtggtcgctg cccgcgttgc catatcccgc tacgcctgcg tcgccgtcat
     3721 agtctgcaaa aatgctgggc ggcgctgtta gcgtcaatcg ttttattgtt acctgccaac
     3781 ctgttgccta tttctatcat ttatctgaac ggaggacggc aggaagatac aattctttcc
     3841 ggaattatgt cgctggcaag tagcaacatt gcggttgcag gaatcgtgtt tatcgccagt
     3901 attctggtcc cgtttactaa agtgatcgtc atgttcactt tactgttgag cattcatttt
     3961 aaatgccagc aaggtttacg cacacgcatt ctgttactgc ggatggtgac ctggattggt
     4021 cgctggtcga tgctcgacct gtttgtcata tctttaacca tgtcgctgat taatcgcgat
     4081 cagatcctcg cttttactat gggaccggct gcgttttatt tcggcgcagc ggtaattttg
     4141 actattcttg ctgtggaatg gctggacagc cgcttacttt gggatgcaca tgagtcagga
     4201 aacgcccgct tcgacgactg aagcgcagat taaaaataaa cgccgtatct cacctttctg
     4261 gctgctgccg ttcatcgcgc taatgattgc cagttggctg atttgggaca gttatcagga
     4321 ccggggtaat accgtcacca tcgactttat gtcggcggat ggtattgttc caggccgtac
     4381 gcctgttcgt tatcagggcg ttgaagtcgg aacagtgcag gacatcagcc tcagcgacga
     4441 tcttcgtaag attgaagtca aggtcagcat caagtccgat atgaaagatg cgctgcgcga
     4501 agagactcag ttctggctgg tgacgccaaa agcatcgttg gcaggtgtct ccgggctgga
     4561 cgccctcgtc ggtgggaact atatcggcat gatgcccggg aaaggtaaag agcaggatca
     4621 ctttgtcgca ctcgataccc aaccgaaata tcggctggac aatggcgatc tgatgatcca
     4681 cctgcaagcc cccgatctcg gttcgctgaa cagcggttca ttggtctatt tccgcaagat
     4741 cccggtggga aaagtctacg actatgccat caatcccaac aagcaaggcg tggtgattga
     4801 tgtcctgatc gagcggcgtt ttaccgatct ggtgaaaaaa ggtagccgtt tctggaacgt
     4861 ttccggcgtt gatgccaacg tcagtatcag tggcgcgaag gtgaaactgg aaagcttggc
     4921 ggcactggtt aacggtgcga ttgccttcga ttcaccagaa gagtcgaaac ctgccgaggc
     4981 ggaagatacc tttggtctgt atgaagatct ggcccacagc cagcgtggcg taataataaa
     5041 actggaactg ccgagtgggg ccggattaac cgccgactcg acgccgttaa tgtatcaggg
     5101 gctggaagtc ggacagctga ctaaactgga tttaaatcct ggtggtaaag tcacagggga
     5161 aatgaccgtt gatcccagcg tcgttaccct gcttcgtgaa aatacccgca tcgaattacg
     5221 caacccgaaa ttatccctta gcgatgctaa tctcagcgcc ctgctgaccg gcaaaacctt
     5281 cgagctggtg cccggcgatg gcgagccacg caaagaattc gttgttgtgc caggcgaaaa
     5341 agcactgctg catgaacctg atgttctgac gctgaccctg accgcgccgg aaagttacgg
     5401 tattgatgcg ggtcagccgc tcattcttca cggcgtgcag gtaggccagg ttatcgatcg
     5461 taaactcacc agcaaaggcg tcacctttac cgtcgccatc gagcctcagc atcgcgaact
     5521 ggtaaaaggc gatagcaaat ttgtcgtcaa cagccgtgtc gatgtgaagg tggggctgga
     5581 tggcgttgag tttctcggtg ccagcgcctc agaatggatc aatggcggga tccgtattct
     5641 gccgggcgat aaaggtgaga tgaaagccag ctatccactg tatgccaatt tggaaaaagc
     5701 gctggagaac agccttagcg atttacccac cacaaccgtg agtttgagtg cagagacgct
     5761 gccggatgtg caggcaggat cggtagtgct gtaccgtaaa tttgaagttg gtgaagtgat
     5821 taccgtgcgt ccgcgagcta acgcgtttga catcgatctg catattaagc cggagtatcg
     5881 caaccttctg accagcaata gcgtgttctg ggcagaaggc ggggcgaaag ttcagctgaa
     5941 tggtagtggt ctgaccgtac aggcatcccc gctctccaga gctctaaagg gagccattag
     6001 cttcgacaac ctcagcggtg ccagcgccag tcagcgtaaa ggcgataagc gtattctgta
     6061 tgcttccgaa acagcggccc gtgcggttgg cgggcagatt acgcttcacg ctttcgatgc
     6121 cggaaaactg gcggtcggga tgccaattcg ctatctcggt attgacatcg ggcaaatcca
     6181 gacgctggat ctgattaccg cacgcaatga agtacaggca aaggcggtac tttacccgga
     6241 atatgtccag acctttgctc gcggtggtac gcgcttctca gtggtcacac cgcaaatttc
     6301 ggcagctggc gttgagcatc ttgatactat cctccagccg tatatcaacg tcgaaccagg
     6361 ccggggcaat cctcgccgcg actttgaatt acaagaggcc accattactg attcgcggta
     6421 cctggatggc ttaagcatta ttgttgaagc gccggaagcc ggttcgttag gcatcggtac
     6481 gcctgtgctg ttccgtggtc tggaagtcgg tacggttaca ggaatgacgc tggggacatt
     6541 gtcagatcgc gtgatgattg cgatgcgcat cagtaaacgc tatcaacacc tggtgcgtaa
     6601 caattccgtc ttctggttgg catcgggtta cagtctggac tttggtctga cgggcggcgt
     6661 agtgaaaacc ggcaccttta accagtttat ccgtggcggc atcgccttcg ccacgcctcc
     6721 gggtacgcca ctggcaccga aagcccagga aggcaaacac ttcctgttgc aggaaagtga
     6781 accgaaagag tggcgtgaat ggggaactgc gcttcccaaa taatgcccac ctcgagcaga
     6841 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     6901 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     6961 aactatatcc ggataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     7021 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg gga
//
