LOCUS       sequence-464780-        6995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA(Delta-ZnR-C) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6995)
  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 6995)
  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: pBEL2879
FEATURES             Location/Qualifiers
     source          1..6995
                     /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(4014..4032)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4028..4075
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4120..4138)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4206..4310
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4311..4379,4380..5171)
                     /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(4529..4548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5342..5930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5831..5850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6084..6101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6116..6255
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6181..6200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6341..6363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6357..6548)
                     /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 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 gcgcagtagt ggaggcgggt ctggtggcgg ctcccgtcgt
      841 cgtcatagtc tgcaaaaatg ctgggcggcg ctgttagcgt caatcgtttt attgttacct
      901 gccaacctgt tgcctatttc tatcatttat ctgaacggag gacggcagga agatacaatt
      961 ctttccggaa ttatgtcgct ggcaagtagc aacattgcgg ttgcaggaat cgtgtttatc
     1021 gccagtattc tggtcccgtt tactaaagtg atcgtcatgt tcactttact gttgagcatt
     1081 cattttaaat gccagcaagg tttacgcaca cgcattctgt tactgcggat ggtgacctgg
     1141 attggtcgct ggtcgatgct cgacctgttt gtcatatctt taaccatgtc gctgattaat
     1201 cgcgatcaga tcctcgcttt tactatggga ccggctgcgt tttatttcgg cgcagcggta
     1261 attttgacta ttcttgctgt ggaatggctg gacagccgct tactttggga tgcacatgag
     1321 tcaggaaacg cccgcttcga cgactgaagc gcagattaaa aataaacgcc gtatctcacc
     1381 tttctggctg ctgccgttca tcgcgctaat gattgccagt tggctgattt gggacagtta
     1441 tcaggaccgg ggtaataccg tcaccatcga ctttatgtcg gcggatggta ttgttccagg
     1501 ccgtacgcct gttcgttatc agggcgttga agtcggaaca gtgcaggaca tcagcctcag
     1561 cgacgatctt cgtaagattg aagtcaaggt cagcatcaag tccgatatga aagatgcgct
     1621 gcgcgaagag actcagttct ggctggtgac gccaaaagca tcgttggcag gtgtctccgg
     1681 gctggacgcc ctcgtcggtg ggaactatat cggcatgatg cccgggaaag gtaaagagca
     1741 ggatcacttt gtcgcactcg atacccaacc gaaatatcgg ctggacaatg gcgatctgat
     1801 gatccacctg caagcccccg atctcggttc gctgaacagc ggttcattgg tctatttccg
     1861 caagatcccg gtgggaaaag tctacgacta tgccatcaat cccaacaagc aaggcgtggt
     1921 gattgatgtc ctgatcgagc ggcgttttac cgatctggtg aaaaaaggta gccgtttctg
     1981 gaacgtttcc ggcgttgatg ccaacgtcag tatcagtggc gcgaaggtga aactggaaag
     2041 cttggcggca ctggttaacg gtgcgattgc cttcgattca ccagaagagt cgaaacctgc
     2101 cgaggcggaa gatacctttg gtctgtatga agatctggcc cacagccagc gtggcgtaat
     2161 aataaaactg gaactgccga gtggggccgg attaaccgcc gactcgacgc cgttaatgta
     2221 tcaggggctg gaagtcggac agctgactaa actggattta aatcctggtg gtaaagtcac
     2281 aggggaaatg accgttgatc ccagcgtcgt taccctgctt cgtgaaaata cccgcatcga
     2341 attacgcaac ccgaaattat cccttagcga tgctaatctc agcgccctgc tgaccggcaa
     2401 aaccttcgag ctggtgcccg gcgatggcga gccacgcaaa gaattcgttg ttgtgccagg
     2461 cgaaaaagca ctgctgcatg aacctgatgt tctgacgctg accctgaccg cgccggaaag
     2521 ttacggtatt gatgcgggtc agccgctcat tcttcacggc gtgcaggtag gccaggttat
     2581 cgatcgtaaa ctcaccagca aaggcgtcac ctttaccgtc gccatcgagc ctcagcatcg
     2641 cgaactggta aaaggcgata gcaaatttgt cgtcaacagc cgtgtcgatg tgaaggtggg
     2701 gctggatggc gttgagtttc tcggtgccag cgcctcagaa tggatcaatg gcgggatccg
     2761 tattctgccg ggcgataaag gtgagatgaa agccagctat ccactgtatg ccaatttgga
     2821 aaaagcgctg gagaacagcc ttagcgattt acccaccaca accgtgagtt tgagtgcaga
     2881 gacgctgccg gatgtgcagg caggatcggt agtgctgtac cgtaaatttg aagttggtga
     2941 agtgattacc gtgcgtccgc gagctaacgc gtttgacatc gatctgcata ttaagccgga
     3001 gtatcgcaac cttctgacca gcaatagcgt gttctgggca gaaggcgggg cgaaagttca
     3061 gctgaatggt agtggtctga ccgtacaggc atccccgctc tccagagctc taaagggagc
     3121 cattagcttc gacaacctca gcggtgccag cgccagtcag cgtaaaggcg ataagcgtat
     3181 tctgtatgct tccgaaacag cggcccgtgc ggttggcggg cagattacgc ttcacgcttt
     3241 cgatgccgga aaactggcgg tcgggatgcc aattcgctat ctcggtattg acatcgggca
     3301 aatccagacg ctggatctga ttaccgcacg caatgaagta caggcaaagg cggtacttta
     3361 cccggaatat gtccagacct ttgctcgcgg tggtacgcgc ttctcagtgg tcacaccgca
     3421 aatttcggca gctggcgttg agcatcttga tactatcctc cagccgtata tcaacgtcga
     3481 accaggccgg ggcaatcctc gccgcgactt tgaattacaa gaggccacca ttactgattc
     3541 gcggtacctg gatggcttaa gcattattgt tgaagcgccg gaagccggtt cgttaggcat
     3601 cggtacgcct gtgctgttcc gtggtctgga agtcggtacg gttacaggaa tgacgctggg
     3661 gacattgtca gatcgcgtga tgattgcgat gcgcatcagt aaacgctatc aacacctggt
     3721 gcgtaacaat tccgtcttct ggttggcatc gggttacagt ctggactttg gtctgacggg
     3781 cggcgtagtg aaaaccggca cctttaacca gtttatccgt ggcggcatcg ccttcgccac
     3841 gcctccgggt acgccactgg caccgaaagc ccaggaaggc aaacacttcc tgttgcagga
     3901 aagtgaaccg aaagagtggc gtgaatgggg aactgcgctt cccaaataat gcccacctcg
     3961 agcagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     4021 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     4081 aggaggaact atatccggat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
     4141 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4201 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4261 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4321 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4381 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4441 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4501 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     4561 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4621 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4681 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4741 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4801 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     4861 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4921 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4981 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5041 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5101 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5161 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5221 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5281 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5341 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5401 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5461 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     5521 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5581 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5641 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5701 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5761 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5821 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5881 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5941 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6001 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6061 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     6121 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     6181 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     6241 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     6301 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     6361 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     6421 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     6481 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     6541 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     6601 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     6661 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     6721 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     6781 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     6841 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     6901 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     6961 ccccgccagc ctagccgggt cctcaacgac aggag
//
