LOCUS       sequence-464729-        7109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of Cys-light LetA(Q180C,R380C) with 
            LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7109)
  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 7109)
  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: pBEL2871
FEATURES             Location/Qualifiers
     source          1..7109
                     /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)"
     RBS             814..836
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4128..4146)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4142..4189
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4234..4252)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4320..4424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4425..4493,4494..5285)
                     /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(4643..4662)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5456..6044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5945..5964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6198..6215
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6230..6369
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6295..6314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6455..6477
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6471..6662)
                     /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 ccgcgtggtg attaccaacg ttgcccgcaa
      241 tgtgacatgc tgtttagcct gcccgagata aattctcatc aaagtgccta ttgtccgcgc
      301 tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc
      361 ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg
      421 ttaggcattc gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc
      481 gatgcgataa cggggtcgat ggtctttttc tctgttattg gtgccccgct cattctggtg
      541 acctccatag cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg
      601 ctgatgcttg agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc
      661 gttgcttcta taaaggtatg cgattatgcc catatccagg ctggtgtcgg cttgttctct
      721 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
      781 tgggagcgat tttatccgca gcgccccgct tgatttgttt aactttaaga aggagatata
      841 catatgacgc gtagggacga gaaacttcgt gtctgcctag ggtgccattt taccggctat
      901 ccagatcagc gtggtcgctg cccgcgttgc catatcccgc tacgcctgcg tcgccgtcat
      961 agtctgcaaa aatcttgggc ggcgctgtta gcgtcaatcg ttttattgtt acctgccaac
     1021 ctgttgccta tttctatcat ttatctgaac ggaggacggc aggaagatac aattctttcc
     1081 ggaattatgt cgctggcaag tagcaacatt gcggttgcag gaatcgtgtt tatcgccagt
     1141 attctggtcc cgtttactaa agtgatcgtc atgttcactt tactgttgag cattcatttt
     1201 aaatctcagc aaggtttacg cacacgcatt ctgttactgc ggatggtgac ctggattggt
     1261 cgctggtcga tgctcgacct gtttgtcata tctttaacca tgtcgctgat taattgcgat
     1321 cagatcctcg cttttactat gggaccggct gcgttttatt tcggcgcagc ggtaattttg
     1381 actattcttg ctgtggaatg gctggacagc cgcttacttt gggatgcaca tgagtcagga
     1441 aacgcccgct tcgacgactg aagcgcagat taaaaataaa cgccgtatct cacctttctg
     1501 gctgctgccg ttcatcgcgc taatgattgc cagttggctg atttgggaca gttatcagga
     1561 ccggggtaat accgtcacca tcgactttat gtcggcggat ggtattgttc caggccgtac
     1621 gcctgttcgt tatcagggcg ttgaagtcgg aacagtgcag gacatcagcc tcagcgacga
     1681 tcttcgtaag attgaagtca aggtcagcat caagtccgat atgaaagatg cgctgcgcga
     1741 agagactcag ttctggctgg tgacgccaaa agcatcgttg gcaggtgtct ccgggctgga
     1801 cgccctcgtc ggtgggaact atatcggcat gatgcccggg aaaggtaaag agcaggatca
     1861 ctttgtcgca ctcgataccc aaccgaaata tcggctggac aatggcgatc tgatgatcca
     1921 cctgcaagcc cccgatctcg gttcgctgaa cagcggttca ttggtctatt tccgcaagat
     1981 cccggtggga aaagtctacg actatgccat caatcccaac aagcaaggcg tggtgattga
     2041 tgtcctgatc gagcggcgtt ttaccgatct ggtgaaaaaa ggtagccgtt tctggaacgt
     2101 ttccggcgtt gatgccaacg tcagtatcag tggcgcgaag gtgaaactgg aaagcttggc
     2161 ggcactggtt aacggtgcga ttgccttcga ttcaccagaa gagtcgaaac ctgccgaggc
     2221 ggaagatacc tttggtctgt atgaagatct ggcccacagc cagcgtggcg taataataaa
     2281 actggaactg ccgagtgggg ccggattaac cgccgactcg acgccgttaa tgtatcaggg
     2341 gctggaagtc ggacagctga ctaaactgga tttaaatcct ggtggtaaag tcacagggga
     2401 aatgaccgtt gatcccagcg tcgttaccct gcttcgtgaa aatacccgca tcgaattacg
     2461 caacccgaaa ttatccctta gcgatgctaa tctcagcgcc ctgctgaccg gcaaaacctt
     2521 cgagctggtg cccggcgatg gcgagccacg caaagaattc gttgttgtgc caggcgaaaa
     2581 agcactgctg catgaacctg atgttctgac gctgaccctg accgcgccgg aaagttacgg
     2641 tattgatgcg ggtcagccgc tcattcttca cggcgtgcag gtaggccagg ttatcgatcg
     2701 taaactcacc agcaaaggcg tcacctttac cgtcgccatc gagcctcagc atcgcgaact
     2761 ggtaaaaggc gatagcaaat ttgtcgtcaa cagccgtgtc gatgtgaagg tggggctgga
     2821 tggcgttgag tttctcggtg ccagcgcctc agaatggatc aatggcggga tccgtattct
     2881 gccgggcgat aaaggtgaga tgaaagccag ctatccactg tatgccaatt tggaaaaagc
     2941 gctggagaac agccttagcg atttacccac cacaaccgtg agtttgagtg cagagacgct
     3001 gccggatgtg caggcaggat cggtagtgct gtaccgtaaa tttgaagttg gtgaagtgat
     3061 taccgtgcgt ccgcgagcta acgcgtttga catcgatctg catattaagc cggagtatcg
     3121 caaccttctg accagcaata gcgtgttctg ggcagaaggc ggggcgaaag ttcagctgaa
     3181 tggtagtggt ctgaccgtac aggcatcccc gctctccaga gctctaaagg gagccattag
     3241 cttcgacaac ctcagcggtg ccagcgccag tcagcgtaaa ggcgataagc gtattctgta
     3301 tgcttccgaa acagcggccc gtgcggttgg cgggcagatt acgcttcacg ctttcgatgc
     3361 cggaaaactg gcggtcggga tgccaattcg ctatctcggt attgacatcg ggcaaatcca
     3421 gacgctggat ctgattaccg cacgcaatga agtacaggca aaggcggtac tttacccgga
     3481 atatgtccag acctttgctc gcggtggtac gcgcttctca gtggtcacac cgcaaatttc
     3541 ggcagctggc gttgagcatc ttgatactat cctccagccg tatatcaacg tcgaaccagg
     3601 ccggggcaat cctcgccgcg actttgaatt acaagaggcc accattactg attcgcggta
     3661 cctggatggc ttaagcatta ttgttgaagc gccggaagcc ggttcgttag gcatcggtac
     3721 gcctgtgctg ttccgtggtc tggaagtcgg tacggttaca ggaatgacgc tggggacatt
     3781 gtcagatcgc gtgatgattg cgatgcgcat cagtaaacgc tatcaacacc tggtgcgtaa
     3841 caattccgtc ttctggttgg catcgggtta cagtctggac tttggtctga cgggcggcgt
     3901 agtgaaaacc ggcaccttta accagtttat ccgtggcggc atcgccttcg ccacgcctcc
     3961 gggtacgcca ctggcaccga aagcccagga aggcaaacac ttcctgttgc aggaaagtga
     4021 accgaaagag tggcgtgaat ggggaactgc gcttcccaaa taatgcccac ctcgagcaga
     4081 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     4141 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     4201 aactatatcc ggataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     4261 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4321 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4381 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4441 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4501 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4561 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4621 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     4681 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4741 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4801 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4861 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4921 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     4981 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5041 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5101 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5161 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5221 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5281 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5341 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5401 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5461 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5521 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5581 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     5641 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5701 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5761 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5821 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5881 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5941 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6001 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6061 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6121 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     6181 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     6241 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     6301 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     6361 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6421 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6481 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     6541 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     6601 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     6661 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     6721 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     6781 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     6841 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     6901 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     6961 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     7021 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     7081 cagcctagcc gggtcctcaa cgacaggag
//
