LOCUS       sequence-464782-        7094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA(PqiA-ZnR) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7094)
  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 7094)
  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: pBEL3017
FEATURES             Location/Qualifiers
     source          1..7094
                     /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(4113..4131)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4127..4174
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4219..4237)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4305..4409
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4410..4478,4479..5270)
                     /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(4628..4647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5441..6029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5930..5949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6183..6200
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6215..6354
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6280..6299)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6440..6462
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6456..6647)
                     /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 tatggctctt aacacaccac aaattacgcc gacaaaaaag
      181 ataacagtga gggcaatcgg cgaggaactg ccgcgtggta agcacatcct ttgcagtcaa
      241 tgtgacatgt tggttgccct tcctcgcctt gaacatggtc aaaaagcggc gtgcccgcgt
      301 tgtggtacga cccttacgga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc
      361 ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg
      421 ttaggcattc gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc
      481 gatgcgataa cggggtcgat ggtctttttc tgcgttattg gtgccccgct cattctggtg
      541 acctccatag cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg
      601 ctgatgcttg agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc
      661 gttgcttcta taaaggtaca ggattatgcc catatccagg ctggtgtcgg cttgttctct
      721 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
      781 tgggagcgat tttatccgca gcgcgagctt cgtcaaccgc ttaagcccgg cgtcacgggt
      841 attcgtcagg gtttgcgttc atgcagttgt tgtactgcca ttcttccagc cgatgagagc
      901 gtctgtcccc gttgtagtac gaaaggttac gtccgtcgtc gtcatagtct gcaaaaatgc
      961 tgggcggcgc tgttagcgtc aatcgtttta ttgttacctg ccaacctgtt gcctatttct
     1021 atcatttatc tgaacggagg acggcaggaa gatacaattc tttccggaat tatgtcgctg
     1081 gcaagtagca acattgcggt tgcaggaatc gtgtttatcg ccagtattct ggtcccgttt
     1141 actaaagtga tcgtcatgtt cactttactg ttgagcattc attttaaatg ccagcaaggt
     1201 ttacgcacac gcattctgtt actgcggatg gtgacctgga ttggtcgctg gtcgatgctc
     1261 gacctgtttg tcatatcttt aaccatgtcg ctgattaatc gcgatcagat cctcgctttt
     1321 actatgggac cggctgcgtt ttatttcggc gcagcggtaa ttttgactat tcttgctgtg
     1381 gaatggctgg acagccgctt actttgggat gcacatgagt caggaaacgc ccgcttcgac
     1441 gactgaagcg cagattaaaa ataaacgccg tatctcacct ttctggctgc tgccgttcat
     1501 cgcgctaatg attgccagtt ggctgatttg ggacagttat caggaccggg gtaataccgt
     1561 caccatcgac tttatgtcgg cggatggtat tgttccaggc cgtacgcctg ttcgttatca
     1621 gggcgttgaa gtcggaacag tgcaggacat cagcctcagc gacgatcttc gtaagattga
     1681 agtcaaggtc agcatcaagt ccgatatgaa agatgcgctg cgcgaagaga ctcagttctg
     1741 gctggtgacg ccaaaagcat cgttggcagg tgtctccggg ctggacgccc tcgtcggtgg
     1801 gaactatatc ggcatgatgc ccgggaaagg taaagagcag gatcactttg tcgcactcga
     1861 tacccaaccg aaatatcggc tggacaatgg cgatctgatg atccacctgc aagcccccga
     1921 tctcggttcg ctgaacagcg gttcattggt ctatttccgc aagatcccgg tgggaaaagt
     1981 ctacgactat gccatcaatc ccaacaagca aggcgtggtg attgatgtcc tgatcgagcg
     2041 gcgttttacc gatctggtga aaaaaggtag ccgtttctgg aacgtttccg gcgttgatgc
     2101 caacgtcagt atcagtggcg cgaaggtgaa actggaaagc ttggcggcac tggttaacgg
     2161 tgcgattgcc ttcgattcac cagaagagtc gaaacctgcc gaggcggaag atacctttgg
     2221 tctgtatgaa gatctggccc acagccagcg tggcgtaata ataaaactgg aactgccgag
     2281 tggggccgga ttaaccgccg actcgacgcc gttaatgtat caggggctgg aagtcggaca
     2341 gctgactaaa ctggatttaa atcctggtgg taaagtcaca ggggaaatga ccgttgatcc
     2401 cagcgtcgtt accctgcttc gtgaaaatac ccgcatcgaa ttacgcaacc cgaaattatc
     2461 ccttagcgat gctaatctca gcgccctgct gaccggcaaa accttcgagc tggtgcccgg
     2521 cgatggcgag ccacgcaaag aattcgttgt tgtgccaggc gaaaaagcac tgctgcatga
     2581 acctgatgtt ctgacgctga ccctgaccgc gccggaaagt tacggtattg atgcgggtca
     2641 gccgctcatt cttcacggcg tgcaggtagg ccaggttatc gatcgtaaac tcaccagcaa
     2701 aggcgtcacc tttaccgtcg ccatcgagcc tcagcatcgc gaactggtaa aaggcgatag
     2761 caaatttgtc gtcaacagcc gtgtcgatgt gaaggtgggg ctggatggcg ttgagtttct
     2821 cggtgccagc gcctcagaat ggatcaatgg cgggatccgt attctgccgg gcgataaagg
     2881 tgagatgaaa gccagctatc cactgtatgc caatttggaa aaagcgctgg agaacagcct
     2941 tagcgattta cccaccacaa ccgtgagttt gagtgcagag acgctgccgg atgtgcaggc
     3001 aggatcggta gtgctgtacc gtaaatttga agttggtgaa gtgattaccg tgcgtccgcg
     3061 agctaacgcg tttgacatcg atctgcatat taagccggag tatcgcaacc ttctgaccag
     3121 caatagcgtg ttctgggcag aaggcggggc gaaagttcag ctgaatggta gtggtctgac
     3181 cgtacaggca tccccgctct ccagagctct aaagggagcc attagcttcg acaacctcag
     3241 cggtgccagc gccagtcagc gtaaaggcga taagcgtatt ctgtatgctt ccgaaacagc
     3301 ggcccgtgcg gttggcgggc agattacgct tcacgctttc gatgccggaa aactggcggt
     3361 cgggatgcca attcgctatc tcggtattga catcgggcaa atccagacgc tggatctgat
     3421 taccgcacgc aatgaagtac aggcaaaggc ggtactttac ccggaatatg tccagacctt
     3481 tgctcgcggt ggtacgcgct tctcagtggt cacaccgcaa atttcggcag ctggcgttga
     3541 gcatcttgat actatcctcc agccgtatat caacgtcgaa ccaggccggg gcaatcctcg
     3601 ccgcgacttt gaattacaag aggccaccat tactgattcg cggtacctgg atggcttaag
     3661 cattattgtt gaagcgccgg aagccggttc gttaggcatc ggtacgcctg tgctgttccg
     3721 tggtctggaa gtcggtacgg ttacaggaat gacgctgggg acattgtcag atcgcgtgat
     3781 gattgcgatg cgcatcagta aacgctatca acacctggtg cgtaacaatt ccgtcttctg
     3841 gttggcatcg ggttacagtc tggactttgg tctgacgggc ggcgtagtga aaaccggcac
     3901 ctttaaccag tttatccgtg gcggcatcgc cttcgccacg cctccgggta cgccactggc
     3961 accgaaagcc caggaaggca aacacttcct gttgcaggaa agtgaaccga aagagtggcg
     4021 tgaatgggga actgcgcttc ccaaataatg cccacctcga gcagatccgg ctgctaacaa
     4081 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     4141 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     4201 attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4261 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4321 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4381 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4441 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4501 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4561 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4621 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     4681 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     4741 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4801 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     4861 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4921 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     4981 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5041 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5101 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5161 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5221 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5281 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5341 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5401 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5461 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5521 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5581 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5641 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5701 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5761 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5821 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5881 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5941 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6001 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6061 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6121 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6181 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     6241 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     6301 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     6361 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6421 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6481 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     6541 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     6601 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     6661 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     6721 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     6781 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     6841 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     6901 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     6961 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     7021 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     7081 ctcaacgaca ggag
//
