LOCUS       sequence-464798-        6989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA(Delta-ZnR-N) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6989)
  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 6989)
  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: pBEL2772
FEATURES             Location/Qualifiers
     source          1..6989
                     /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(4008..4026)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4022..4069
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4114..4132)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4200..4304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4305..4373,4374..5165)
                     /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(4523..4542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5336..5924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5825..5844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6078..6095
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6110..6249
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6175..6194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6335..6357
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6351..6542)
                     /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 ccgcgtagta gtggaggcgg gtctgacggg
      241 cgcgactggt cgctaacgcg cctggcggca atggccttca ccatgctgtt gcttatgccg
      301 tttgcctggg gcgaaccgct gttgcatatc tggctgttag gcattcgtat tgacgccaac
      361 gttatgcaag gcatctggca aatgaccaaa cagggcgatg cgataacggg gtcgatggtc
      421 tttttctgcg ttattggtgc cccgctcatt ctggtgacct ccatagctta tttatggttt
      481 ggtaaccgac tgggaatgaa tttacgtccg gtactgctga tgcttgagcg acttaaagag
      541 tgggtaatgc tcgatatcta cctggtcggc attggcgttg cttctataaa ggtacaggat
      601 tatgcccata tccaggctgg tgtcggcttg ttctcttttg tggcgttggt gattttaacg
      661 acggtgacgt tgtcacatct taatgtcgaa gaactgtggg agcgatttta tccgcagcgc
      721 cccgctacgc gtagggacga gaaacttcgt gtctgcctag ggtgccattt taccggctat
      781 ccagatcagc gtggtcgctg cccgcgttgc catatcccgc tacgcctgcg tcgccgtcat
      841 agtctgcaaa aatgctgggc ggcgctgtta gcgtcaatcg ttttattgtt acctgccaac
      901 ctgttgccta tttctatcat ttatctgaac ggaggacggc aggaagatac aattctttcc
      961 ggaattatgt cgctggcaag tagcaacatt gcggttgcag gaatcgtgtt tatcgccagt
     1021 attctggtcc cgtttactaa agtgatcgtc atgttcactt tactgttgag cattcatttt
     1081 aaatgccagc aaggtttacg cacacgcatt ctgttactgc ggatggtgac ctggattggt
     1141 cgctggtcga tgctcgacct gtttgtcata tctttaacca tgtcgctgat taatcgcgat
     1201 cagatcctcg cttttactat gggaccggct gcgttttatt tcggcgcagc ggtaattttg
     1261 actattcttg ctgtggaatg gctggacagc cgcttacttt gggatgcaca tgagtcagga
     1321 aacgcccgct tcgacgactg aagcgcagat taaaaataaa cgccgtatct cacctttctg
     1381 gctgctgccg ttcatcgcgc taatgattgc cagttggctg atttgggaca gttatcagga
     1441 ccggggtaat accgtcacca tcgactttat gtcggcggat ggtattgttc caggccgtac
     1501 gcctgttcgt tatcagggcg ttgaagtcgg aacagtgcag gacatcagcc tcagcgacga
     1561 tcttcgtaag attgaagtca aggtcagcat caagtccgat atgaaagatg cgctgcgcga
     1621 agagactcag ttctggctgg tgacgccaaa agcatcgttg gcaggtgtct ccgggctgga
     1681 cgccctcgtc ggtgggaact atatcggcat gatgcccggg aaaggtaaag agcaggatca
     1741 ctttgtcgca ctcgataccc aaccgaaata tcggctggac aatggcgatc tgatgatcca
     1801 cctgcaagcc cccgatctcg gttcgctgaa cagcggttca ttggtctatt tccgcaagat
     1861 cccggtggga aaagtctacg actatgccat caatcccaac aagcaaggcg tggtgattga
     1921 tgtcctgatc gagcggcgtt ttaccgatct ggtgaaaaaa ggtagccgtt tctggaacgt
     1981 ttccggcgtt gatgccaacg tcagtatcag tggcgcgaag gtgaaactgg aaagcttggc
     2041 ggcactggtt aacggtgcga ttgccttcga ttcaccagaa gagtcgaaac ctgccgaggc
     2101 ggaagatacc tttggtctgt atgaagatct ggcccacagc cagcgtggcg taataataaa
     2161 actggaactg ccgagtgggg ccggattaac cgccgactcg acgccgttaa tgtatcaggg
     2221 gctggaagtc ggacagctga ctaaactgga tttaaatcct ggtggtaaag tcacagggga
     2281 aatgaccgtt gatcccagcg tcgttaccct gcttcgtgaa aatacccgca tcgaattacg
     2341 caacccgaaa ttatccctta gcgatgctaa tctcagcgcc ctgctgaccg gcaaaacctt
     2401 cgagctggtg cccggcgatg gcgagccacg caaagaattc gttgttgtgc caggcgaaaa
     2461 agcactgctg catgaacctg atgttctgac gctgaccctg accgcgccgg aaagttacgg
     2521 tattgatgcg ggtcagccgc tcattcttca cggcgtgcag gtaggccagg ttatcgatcg
     2581 taaactcacc agcaaaggcg tcacctttac cgtcgccatc gagcctcagc atcgcgaact
     2641 ggtaaaaggc gatagcaaat ttgtcgtcaa cagccgtgtc gatgtgaagg tggggctgga
     2701 tggcgttgag tttctcggtg ccagcgcctc agaatggatc aatggcggga tccgtattct
     2761 gccgggcgat aaaggtgaga tgaaagccag ctatccactg tatgccaatt tggaaaaagc
     2821 gctggagaac agccttagcg atttacccac cacaaccgtg agtttgagtg cagagacgct
     2881 gccggatgtg caggcaggat cggtagtgct gtaccgtaaa tttgaagttg gtgaagtgat
     2941 taccgtgcgt ccgcgagcta acgcgtttga catcgatctg catattaagc cggagtatcg
     3001 caaccttctg accagcaata gcgtgttctg ggcagaaggc ggggcgaaag ttcagctgaa
     3061 tggtagtggt ctgaccgtac aggcatcccc gctctccaga gctctaaagg gagccattag
     3121 cttcgacaac ctcagcggtg ccagcgccag tcagcgtaaa ggcgataagc gtattctgta
     3181 tgcttccgaa acagcggccc gtgcggttgg cgggcagatt acgcttcacg ctttcgatgc
     3241 cggaaaactg gcggtcggga tgccaattcg ctatctcggt attgacatcg ggcaaatcca
     3301 gacgctggat ctgattaccg cacgcaatga agtacaggca aaggcggtac tttacccgga
     3361 atatgtccag acctttgctc gcggtggtac gcgcttctca gtggtcacac cgcaaatttc
     3421 ggcagctggc gttgagcatc ttgatactat cctccagccg tatatcaacg tcgaaccagg
     3481 ccggggcaat cctcgccgcg actttgaatt acaagaggcc accattactg attcgcggta
     3541 cctggatggc ttaagcatta ttgttgaagc gccggaagcc ggttcgttag gcatcggtac
     3601 gcctgtgctg ttccgtggtc tggaagtcgg tacggttaca ggaatgacgc tggggacatt
     3661 gtcagatcgc gtgatgattg cgatgcgcat cagtaaacgc tatcaacacc tggtgcgtaa
     3721 caattccgtc ttctggttgg catcgggtta cagtctggac tttggtctga cgggcggcgt
     3781 agtgaaaacc ggcaccttta accagtttat ccgtggcggc atcgccttcg ccacgcctcc
     3841 gggtacgcca ctggcaccga aagcccagga aggcaaacac ttcctgttgc aggaaagtga
     3901 accgaaagag tggcgtgaat ggggaactgc gcttcccaaa taatgcccac ctcgagcaga
     3961 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     4021 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     4081 aactatatcc ggataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     4141 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4201 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4261 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4321 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4381 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4441 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4501 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     4561 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4621 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4681 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4741 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4801 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     4861 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     4921 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     4981 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5041 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5101 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5161 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5221 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5281 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5341 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5401 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5461 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5521 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5581 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5641 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5701 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5761 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5821 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5881 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5941 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6001 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6061 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     6121 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     6181 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     6241 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6301 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6361 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     6421 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     6481 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     6541 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     6601 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     6661 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     6721 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     6781 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     6841 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     6901 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     6961 cagcctagcc gggtcctcaa cgacaggag
//
