LOCUS       sequence-464783-        7073 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetB(R63N) with LetA.
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: pBEL3159
FEATURES             Location/Qualifiers
     source          1..7073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..36)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        104..208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(209..277,278..1069)
                     /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(427..446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1729..1748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1982..1999
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2014..2153
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2079..2098)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2239..2261
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2255..2446)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2955..2974
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2955..2973
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3005..3027
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(6985..7003)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6999..7046
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
     2101 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     2161 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     2221 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2281 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     2341 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     2401 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
     2461 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
     2521 tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg
     2581 cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta
     2641 atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     2701 taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga
     2761 ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct
     2821 cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc
     2881 tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatct
     2941 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaaa
     3001 taattttgtt taactttaag aaggagatat acatatggct cttaacacac cacaaattac
     3061 gccgacaaaa aagataacag tgagggcaat cggcgaggaa ctgccgcgtg gtgattacca
     3121 acgttgcccg caatgtgaca tgctgtttag cctgcccgag ataaattctc atcaaagtgc
     3181 ctattgtccg cgctgtcagg caaaaattcg tgacgggcgc gactggtcgc taacgcgcct
     3241 ggcggcaatg gccttcacca tgctgttgct tatgccgttt gcctggggcg aaccgctgtt
     3301 gcatatctgg ctgttaggca ttcgtattga cgccaacgtt atgcaaggca tctggcaaat
     3361 gaccaaacag ggcgatgcga taacggggtc gatggtcttt ttctgcgtta ttggtgcccc
     3421 gctcattctg gtgacctcca tagcttattt atggtttggt aaccgactgg gaatgaattt
     3481 acgtccggta ctgctgatgc ttgagcgact taaagagtgg gtaatgctcg atatctacct
     3541 ggtcggcatt ggcgttgctt ctataaaggt acaggattat gcccatatcc aggctggtgt
     3601 cggcttgttc tcttttgtgg cgttggtgat tttaacgacg gtgacgttgt cacatcttaa
     3661 tgtcgaagaa ctgtgggagc gattttatcc gcagcgcccc gctacgcgta gggacgagaa
     3721 acttcgtgtc tgcctagggt gccattttac cggctatcca gatcagcgtg gtcgctgccc
     3781 gcgttgccat atcccgctac gcctgcgtcg ccgtcatagt ctgcaaaaat gctgggcggc
     3841 gctgttagcg tcaatcgttt tattgttacc tgccaacctg ttgcctattt ctatcattta
     3901 tctgaacgga ggacggcagg aagatacaat tctttccgga attatgtcgc tggcaagtag
     3961 caacattgcg gttgcaggaa tcgtgtttat cgccagtatt ctggtcccgt ttactaaagt
     4021 gatcgtcatg ttcactttac tgttgagcat tcattttaaa tgccagcaag gtttacgcac
     4081 acgcattctg ttactgcgga tggtgacctg gattggtcgc tggtcgatgc tcgacctgtt
     4141 tgtcatatct ttaaccatgt cgctgattaa tcgcgatcag atcctcgctt ttactatggg
     4201 accggctgcg ttttatttcg gcgcagcggt aattttgact attcttgctg tggaatggct
     4261 ggacagccgc ttactttggg atgcacatga gtcaggaaac gcccgcttcg acgactgaag
     4321 cgcagattaa aaataaacgc cgtatctcac ctttctggct gctgccgttc atcgcgctaa
     4381 tgattgccag ttggctgatt tgggacagtt atcaggaccg gggtaatacc gtcaccatcg
     4441 actttatgtc ggcggatggt attgttccag gcaacacgcc tgttcgttat cagggcgttg
     4501 aagtcggaac agtgcaggac atcagcctca gcgacgatct tcgtaagatt gaagtcaagg
     4561 tcagcatcaa gtccgatatg aaagatgcgc tgcgcgaaga gactcagttc tggctggtga
     4621 cgccaaaagc atcgttggca ggtgtctccg ggctggacgc cctcgtcggt gggaactata
     4681 tcggcatgat gcccgggaaa ggtaaagagc aggatcactt tgtcgcactc gatacccaac
     4741 cgaaatatcg gctggacaat ggcgatctga tgatccacct gcaagccccc gatctcggtt
     4801 cgctgaacag cggttcattg gtctatttcc gcaagatccc ggtgggaaaa gtctacgact
     4861 atgccatcaa tcccaacaag caaggcgtgg tgattgatgt cctgatcgag cggcgtttta
     4921 ccgatctggt gaaaaaaggt agccgtttct ggaacgtttc cggcgttgat gccaacgtca
     4981 gtatcagtgg cgcgaaggtg aaactggaaa gcttggcggc actggttaac ggtgcgattg
     5041 ccttcgattc accagaagag tcgaaacctg ccgaggcgga agataccttt ggtctgtatg
     5101 aagatctggc ccacagccag cgtggcgtaa taataaaact ggaactgccg agtggggccg
     5161 gattaaccgc cgactcgacg ccgttaatgt atcaggggct ggaagtcgga cagctgacta
     5221 aactggattt aaatcctggt ggtaaagtca caggggaaat gaccgttgat cccagcgtcg
     5281 ttaccctgct tcgtgaaaat acccgcatcg aattacgcaa cccgaaatta tcccttagcg
     5341 atgctaatct cagcgccctg ctgaccggca aaaccttcga gctggtgccc ggcgatggcg
     5401 agccacgcaa agaattcgtt gttgtgccag gcgaaaaagc actgctgcat gaacctgatg
     5461 ttctgacgct gaccctgacc gcgccggaaa gttacggtat tgatgcgggt cagccgctca
     5521 ttcttcacgg cgtgcaggta ggccaggtta tcgatcgtaa actcaccagc aaaggcgtca
     5581 cctttaccgt cgccatcgag cctcagcatc gcgaactggt aaaaggcgat agcaaatttg
     5641 tcgtcaacag ccgtgtcgat gtgaaggtgg ggctggatgg cgttgagttt ctcggtgcca
     5701 gcgcctcaga atggatcaat ggcgggatcc gtattctgcc gggcgataaa ggtgagatga
     5761 aagccagcta tccactgtat gccaatttgg aaaaagcgct ggagaacagc cttagcgatt
     5821 tacccaccac aaccgtgagt ttgagtgcag agacgctgcc ggatgtgcag gcaggatcgg
     5881 tagtgctgta ccgtaaattt gaagttggtg aagtgattac cgtgcgtccg cgagctaacg
     5941 cgtttgacat cgatctgcat attaagccgg agtatcgcaa ccttctgacc agcaatagcg
     6001 tgttctgggc agaaggcggg gcgaaagttc agctgaatgg tagtggtctg accgtacagg
     6061 catccccgct ctccagagct ctaaagggag ccattagctt cgacaacctc agcggtgcca
     6121 gcgccagtca gcgtaaaggc gataagcgta ttctgtatgc ttccgaaaca gcggcccgtg
     6181 cggttggcgg gcagattacg cttcacgctt tcgatgccgg aaaactggcg gtcgggatgc
     6241 caattcgcta tctcggtatt gacatcgggc aaatccagac gctggatctg attaccgcac
     6301 gcaatgaagt acaggcaaag gcggtacttt acccggaata tgtccagacc tttgctcgcg
     6361 gtggtacgcg cttctcagtg gtcacaccgc aaatttcggc agctggcgtt gagcatcttg
     6421 atactatcct ccagccgtat atcaacgtcg aaccaggccg gggcaatcct cgccgcgact
     6481 ttgaattaca agaggccacc attactgatt cgcggtacct ggatggctta agcattattg
     6541 ttgaagcgcc ggaagccggt tcgttaggca tcggtacgcc tgtgctgttc cgtggtctgg
     6601 aagtcggtac ggttacagga atgacgctgg ggacattgtc agatcgcgtg atgattgcga
     6661 tgcgcatcag taaacgctat caacacctgg tgcgtaacaa ttccgtcttc tggttggcat
     6721 cgggttacag tctggacttt ggtctgacgg gcggcgtagt gaaaaccggc acctttaacc
     6781 agtttatccg tggcggcatc gccttcgcca cgcctccggg tacgccactg gcaccgaaag
     6841 cccaggaagg caaacacttc ctgttgcagg aaagtgaacc gaaagagtgg cgtgaatggg
     6901 gaactgcgct tcccaaataa tgcccacctc gagcagatcc ggctgctaac aaagcccgaa
     6961 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     7021 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga taa
//
