LOCUS       sequence-464815-        7004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA(Delta-2-24) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7004)
  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 7004)
  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: pBEL2472
FEATURES             Location/Qualifiers
     source          1..7004
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..81
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             112..134
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4023..4041)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4037..4084
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4129..4147)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4215..4319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4320..4388,4389..5180)
                     /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(4538..4557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5351..5939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5840..5859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6093..6110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6125..6264
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6190..6209)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6350..6372
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6366..6557)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatctcga tcccgcgaaa
       61 ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa ttttgtttaa
      121 ctttaagaag gagatataca tatgcgtggt gattaccaac gttgcccgca atgtgacatg
      181 ctgtttagcc tgcccgagat aaattctcat caaagtgcct attgtccgcg ctgtcaggca
      241 aaaattcgtg acgggcgcga ctggtcgcta acgcgcctgg cggcaatggc cttcaccatg
      301 ctgttgctta tgccgtttgc ctggggcgaa ccgctgttgc atatctggct gttaggcatt
      361 cgtattgacg ccaacgttat gcaaggcatc tggcaaatga ccaaacaggg cgatgcgata
      421 acggggtcga tggtcttttt ctgcgttatt ggtgccccgc tcattctggt gacctccata
      481 gcttatttat ggtttggtaa ccgactggga atgaatttac gtccggtact gctgatgctt
      541 gagcgactta aagagtgggt aatgctcgat atctacctgg tcggcattgg cgttgcttct
      601 ataaaggtac aggattatgc ccatatccag gctggtgtcg gcttgttctc ttttgtggcg
      661 ttggtgattt taacgacggt gacgttgtca catcttaatg tcgaagaact gtgggagcga
      721 ttttatccgc agcgccccgc tacgcgtagg gacgagaaac ttcgtgtctg cctagggtgc
      781 cattttaccg gctatccaga tcagcgtggt cgctgcccgc gttgccatat cccgctacgc
      841 ctgcgtcgcc gtcatagtct gcaaaaatgc tgggcggcgc tgttagcgtc aatcgtttta
      901 ttgttacctg ccaacctgtt gcctatttct atcatttatc tgaacggagg acggcaggaa
      961 gatacaattc tttccggaat tatgtcgctg gcaagtagca acattgcggt tgcaggaatc
     1021 gtgtttatcg ccagtattct ggtcccgttt actaaagtga tcgtcatgtt cactttactg
     1081 ttgagcattc attttaaatg ccagcaaggt ttacgcacac gcattctgtt actgcggatg
     1141 gtgacctgga ttggtcgctg gtcgatgctc gacctgtttg tcatatcttt aaccatgtcg
     1201 ctgattaatc gcgatcagat cctcgctttt actatgggac cggctgcgtt ttatttcggc
     1261 gcagcggtaa ttttgactat tcttgctgtg gaatggctgg acagccgctt actttgggat
     1321 gcacatgagt caggaaacgc ccgcttcgac gactgaagcg cagattaaaa ataaacgccg
     1381 tatctcacct ttctggctgc tgccgttcat cgcgctaatg attgccagtt ggctgatttg
     1441 ggacagttat caggaccggg gtaataccgt caccatcgac tttatgtcgg cggatggtat
     1501 tgttccaggc cgtacgcctg ttcgttatca gggcgttgaa gtcggaacag tgcaggacat
     1561 cagcctcagc gacgatcttc gtaagattga agtcaaggtc agcatcaagt ccgatatgaa
     1621 agatgcgctg cgcgaagaga ctcagttctg gctggtgacg ccaaaagcat cgttggcagg
     1681 tgtctccggg ctggacgccc tcgtcggtgg gaactatatc ggcatgatgc ccgggaaagg
     1741 taaagagcag gatcactttg tcgcactcga tacccaaccg aaatatcggc tggacaatgg
     1801 cgatctgatg atccacctgc aagcccccga tctcggttcg ctgaacagcg gttcattggt
     1861 ctatttccgc aagatcccgg tgggaaaagt ctacgactat gccatcaatc ccaacaagca
     1921 aggcgtggtg attgatgtcc tgatcgagcg gcgttttacc gatctggtga aaaaaggtag
     1981 ccgtttctgg aacgtttccg gcgttgatgc caacgtcagt atcagtggcg cgaaggtgaa
     2041 actggaaagc ttggcggcac tggttaacgg tgcgattgcc ttcgattcac cagaagagtc
     2101 gaaacctgcc gaggcggaag atacctttgg tctgtatgaa gatctggccc acagccagcg
     2161 tggcgtaata ataaaactgg aactgccgag tggggccgga ttaaccgccg actcgacgcc
     2221 gttaatgtat caggggctgg aagtcggaca gctgactaaa ctggatttaa atcctggtgg
     2281 taaagtcaca ggggaaatga ccgttgatcc cagcgtcgtt accctgcttc gtgaaaatac
     2341 ccgcatcgaa ttacgcaacc cgaaattatc ccttagcgat gctaatctca gcgccctgct
     2401 gaccggcaaa accttcgagc tggtgcccgg cgatggcgag ccacgcaaag aattcgttgt
     2461 tgtgccaggc gaaaaagcac tgctgcatga acctgatgtt ctgacgctga ccctgaccgc
     2521 gccggaaagt tacggtattg atgcgggtca gccgctcatt cttcacggcg tgcaggtagg
     2581 ccaggttatc gatcgtaaac tcaccagcaa aggcgtcacc tttaccgtcg ccatcgagcc
     2641 tcagcatcgc gaactggtaa aaggcgatag caaatttgtc gtcaacagcc gtgtcgatgt
     2701 gaaggtgggg ctggatggcg ttgagtttct cggtgccagc gcctcagaat ggatcaatgg
     2761 cgggatccgt attctgccgg gcgataaagg tgagatgaaa gccagctatc cactgtatgc
     2821 caatttggaa aaagcgctgg agaacagcct tagcgattta cccaccacaa ccgtgagttt
     2881 gagtgcagag acgctgccgg atgtgcaggc aggatcggta gtgctgtacc gtaaatttga
     2941 agttggtgaa gtgattaccg tgcgtccgcg agctaacgcg tttgacatcg atctgcatat
     3001 taagccggag tatcgcaacc ttctgaccag caatagcgtg ttctgggcag aaggcggggc
     3061 gaaagttcag ctgaatggta gtggtctgac cgtacaggca tccccgctct ccagagctct
     3121 aaagggagcc attagcttcg acaacctcag cggtgccagc gccagtcagc gtaaaggcga
     3181 taagcgtatt ctgtatgctt ccgaaacagc ggcccgtgcg gttggcgggc agattacgct
     3241 tcacgctttc gatgccggaa aactggcggt cgggatgcca attcgctatc tcggtattga
     3301 catcgggcaa atccagacgc tggatctgat taccgcacgc aatgaagtac aggcaaaggc
     3361 ggtactttac ccggaatatg tccagacctt tgctcgcggt ggtacgcgct tctcagtggt
     3421 cacaccgcaa atttcggcag ctggcgttga gcatcttgat actatcctcc agccgtatat
     3481 caacgtcgaa ccaggccggg gcaatcctcg ccgcgacttt gaattacaag aggccaccat
     3541 tactgattcg cggtacctgg atggcttaag cattattgtt gaagcgccgg aagccggttc
     3601 gttaggcatc ggtacgcctg tgctgttccg tggtctggaa gtcggtacgg ttacaggaat
     3661 gacgctgggg acattgtcag atcgcgtgat gattgcgatg cgcatcagta aacgctatca
     3721 acacctggtg cgtaacaatt ccgtcttctg gttggcatcg ggttacagtc tggactttgg
     3781 tctgacgggc ggcgtagtga aaaccggcac ctttaaccag tttatccgtg gcggcatcgc
     3841 cttcgccacg cctccgggta cgccactggc accgaaagcc caggaaggca aacacttcct
     3901 gttgcaggaa agtgaaccga aagagtggcg tgaatgggga actgcgcttc ccaaataatg
     3961 cccacctcga gcagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     4021 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     4081 tttgctgaaa ggaggaacta tatccggata attcttgaag acgaaagggc ctcgtgatac
     4141 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4201 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4261 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4321 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4381 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4441 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4501 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4561 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4621 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4681 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4741 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4801 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4861 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4921 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4981 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5041 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5101 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5161 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5221 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5281 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5341 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5401 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5461 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5521 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5581 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5641 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5701 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5761 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5821 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5881 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5941 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6001 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6061 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6121 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     6181 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     6241 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     6301 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     6361 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     6421 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     6481 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     6541 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     6601 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     6661 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     6721 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     6781 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     6841 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     6901 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     6961 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggag
//
