LOCUS       sequence-333955-        7070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetic complementation plasmid of LetA-[LetB PLL6->3] mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7070)
  AUTHORS   Vieni C, Coudray N, Isom GL, Bhabha G, Ekiert DC
  TITLE     Role of Ring6 in the Function of the E. coli MCE Protein LetB.
  JOURNAL   J Mol Biol. 2022 Jan 22;434(7):167463. doi: 
            10.1016/j.jmb.2022.167463.
  PUBMED    35077766
REFERENCE   2  (bases 1 to 7070)
  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: "J Mol 
            Biol."; date: "2022-01-22"; volume: "434(7):167463. doi"; pages: " 
            10.1016/j.jmb.2022.167463"
COMMENT     Sequence Label: pBEL2200
FEATURES             Location/Qualifiers
     source          1..7070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     439..458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     692..709
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    724..863
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(789..808)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     949..971
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(965..1156)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1665..1684
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1665..1683
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1715..1737
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(5692..5710)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5706..5753
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(5798..5816)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5884..5988
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5989..6057,6058..6849)
                     /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(6207..6226)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7020..7070,1..538)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
       61 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
      121 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      181 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      241 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      301 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      361 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      421 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      481 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      541 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      601 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      661 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      721 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
      781 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
      841 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
      901 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
      961 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     1021 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     1081 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     1141 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     1201 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     1261 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     1321 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     1381 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     1441 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     1501 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     1561 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     1621 gtggccagga cccaacgctg cccgagatct cgatcccgcg aaattaatac gactcactat
     1681 agggagacca caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat
     1741 acatatggct cttaacacac cacaaattac gccgacaaaa aagataacag tgagggcaat
     1801 cggcgaggaa ctgccgcgtg gtgattacca acgttgcccg caatgtgaca tgctgtttag
     1861 cctgcccgag ataaattctc atcaaagtgc ctattgtccg cgctgtcagg caaaaattcg
     1921 tgacgggcgc gactggtcgc taacgcgcct ggcggcaatg gccttcacca tgctgttgct
     1981 tatgccgttt gcctggggcg aaccgctgtt gcatatctgg ctgttaggca ttcgtattga
     2041 cgccaacgtt atgcaaggca tctggcaaat gaccaaacag ggcgatgcga taacggggtc
     2101 gatggtcttt ttctgcgtta ttggtgcccc gctcattctg gtgacctcca tagcttattt
     2161 atggtttggt aaccgactgg gaatgaattt acgtccggta ctgctgatgc ttgagcgact
     2221 taaagagtgg gtaatgctcg atatctacct ggtcggcatt ggcgttgctt ctataaaggt
     2281 acaggattat gcccatatcc aggctggtgt cggcttgttc tcttttgtgg cgttggtgat
     2341 tttaacgacg gtgacgttgt cacatcttaa tgtcgaagaa ctgtgggagc gattttatcc
     2401 gcagcgcccc gctacgcgta gggacgagaa acttcgtgtc tgtcttgggt gccattttac
     2461 cggctatcca gatcagcgtg gtcgctgccc gcgttgccat atcccgctac gcctgcgtcg
     2521 ccgtcatagt ctgcaaaaat gctgggcggc gctgttagcg tcaatcgttt tattgttacc
     2581 tgccaacctg ttgcctattt ctatcattta tctgaacgga ggacggcagg aagatacaat
     2641 tctttccgga attatgtcgc tggcaagtag caacattgcg gttgcaggaa tcgtgtttat
     2701 cgccagtatt ctggtaccgt ttactaaagt gatcgtcatg ttcactttac tgttgagcat
     2761 tcattttaaa tgccagcaag gtttacgcac acgcattctg ttactgcgga tggtgacctg
     2821 gattggtcgc tggtcgatgc tcgacctgtt tgtcatatct ttaaccatgt cgctgattaa
     2881 tcgcgatcag atcctcgctt ttactatggg accggctgcg ttttatttcg gcgcagcggt
     2941 aattttgact attcttgctg tggaatggct ggacagccgc ttactttggg atgcacatga
     3001 gtcaggaaac gcccgcttcg acgactgaag cgcagattaa aaataaacgc cgtatctcac
     3061 ctttctggct gctgccgttc atcgcgctaa tgattgccag ttggctgatt tgggacagtt
     3121 atcaggaccg gggtaatacc gtcaccatcg actttatgtc ggcggatggt attgttccag
     3181 gccgtacgcc tgttcgttat cagggcgttg aagtcggaac agtgcaggat atcagcctca
     3241 gcgacgatct tcgtaagatt gaagtcaagg tcagcatcaa gtccgatatg aaagatgcgc
     3301 tgcgcgaaga gactcagttc tggctggtga cgccaaaagc atcgttggca ggtgtctccg
     3361 ggctggacgc cctcgtcggt gggaactata tcggcatgat gccgggtaaa ggtaaagagc
     3421 aggatcactt tgtcgcactc gatacccaac cgaaatatcg gctggacaat ggcgatctga
     3481 tgatccacct gcaagccccc gatctcggtt cgctgaacag cggttcattg gtctatttcc
     3541 gcaagatccc ggtgggaaaa gtctacgact atgccatcaa tcccaacaag caaggcgtgg
     3601 tgattgatgt cctgatcgag cggcgtttta ccgatctggt gaaaaaaggt agccgtttct
     3661 ggaacgtttc cggcgttgat gccaacgtca gtatcagtgg cgcgaaggtg aaactggaaa
     3721 gcctggcggc actggttaac ggtgcgattg ccttcgattc accagaagag tcgaaacctg
     3781 ccgaggcgga agataccttt ggtctgtatg aagatctggc ccacagccag cgtggcgtaa
     3841 taataaaact ggaactgccg agtggggccg gattaaccgc cgactcgacg ccgttaatgt
     3901 atcaggggct ggaagtcgga cagctgacta aactggattt aaatcctggt ggtaaagtca
     3961 caggggaaat gaccgttgat cccagcgtcg ttaccctgct tcgtgaaaat acccgcatcg
     4021 aattacgcaa cccgaaatta tcccttagcg atgctaatct cagcgccctg ctgaccggca
     4081 aaaccttcga gctggtgccc ggcgatggcg agccacgcaa agagttcgtt gttgtgccag
     4141 gcgaaaaagc actgctgcat gaacctgatg ttctgacgct gaccctgacc gcgccggaaa
     4201 gttacggtat tgatgcgggt cagccgctca ttcttcacgg cgtgcaggta ggccaggtta
     4261 tcgatcgtaa actcaccagc aaaggcgtca cctttaccgt cgccatcgag cctcagcatc
     4321 gcgaactggt aaaaggcgat agcaaatttg tcgtcaacag ccgtgtcgat gtgaaggtgg
     4381 ggctggatgg cgttgagttt ctcggtgcca gcgcctcaga atggatcaat ggcgggatac
     4441 gtattctgcc gggcgataaa ggtgagatga aagccagcta tccactgtat gccaatttgg
     4501 aaaaagcgct ggagaacagc cttagcgatt tacccaccac aaccgtgagt ttgagtgcag
     4561 agacgctgcc ggatgtgcag gcaggatcgg tagtgctgta ccgtaaattt gaagttggtg
     4621 aagtgattac cgtgcgtccg cgagctaacg cgtttgatat cgatctgcat attaagccgg
     4681 agtatcgcaa ccttctgacc agcaatagcg tgttctgggc agaaggcggg gcgaaagttc
     4741 agctgaatgg tagtggtctg accgtacagg catccccgct ctccagagca ttaaagggag
     4801 ccattagctt cgacaacctc agcggtgcca gcgccagtca gcgtaaaggc gataagcgta
     4861 ttctgtatgc ttccgaaaca gcggcccgtg cggttggcgg gcagattacg cttcacgctt
     4921 tcgatgccgg aaaactggcg gtcgggatgc caattcgcta tctcggtatt gatatcgggc
     4981 aaatccagac gctggatctg attaccgcac gcaatgaagt acaggcaaag gcggtacttt
     5041 acccggaata tgtccagacc tttgctcgcg gtggtacgcg cttctcagtg gtcacaccga
     5101 aattatccct tagcgatgct aatctcagcg ccctgctgac cccgtatatc aacgtcgaac
     5161 caggccgggg caatcctcgc cgcgactttg aattacaaga ggccaccatt actgattcgc
     5221 gttacctgga tggcttaagc attattgttg aagcgccgga agccggttcg ttaggcatcg
     5281 gtacgcctgt gctgttccgt ggtctggaag tcggtacggt tacaggaatg acgctgggga
     5341 cattgtcaga tcgcgtgatg attgcgatgc gcatcagtaa acgctatcaa cacctggtgc
     5401 gtaacaattc cgtcttctgg ttggcatcgg gttacagtct ggactttggt ctgacgggcg
     5461 gcgtagtgaa aaccggcacc tttaaccagt ttatccgtgg cggcatcgcc ttcgccacgc
     5521 ctccgggtac gccactggca ccgaaagccc aggaaggcaa acacttcctg ttgcaggaaa
     5581 gtgaaccgaa agagtggcgt gaatggggaa ctgcgcttcc caaataatgc ccacctcgag
     5641 cagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     5701 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     5761 gaggaactat atccggataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     5821 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5881 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5941 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     6001 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6061 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6121 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6181 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     6241 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6301 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6361 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6421 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6481 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     6541 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6601 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6661 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6721 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6781 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6841 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6901 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6961 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7021 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
//
