LOCUS       sequence-333950-        8664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expresson of LetA-[LetB PLL6->1] operon from E. coli. N-terminal
            his tag on LetA, araBAD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8664)
  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 8664)
  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: pBEL2196
FEATURES             Location/Qualifiers
     source          1..8664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(32..65)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     237..254
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     420..442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     580..599
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(779..1657)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1684..1968
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1891..1910
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2005..2028
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             2041..2061
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     primer_bind     complement(6058..6075)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6228..6314
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6406..6433
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6452..6543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6544..6612,6613..7404)
                     /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(6762..6781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7446..7901
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7533..7552)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7743..7764
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      8012..8599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8500..8519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttggctggac gtaaactcct cttcagacct aataacttcg tatagcatac attatacgaa
       61 gttatattaa gggttattga atatgatcgg aattgggctg caggaattcg atagcttggc
      121 tgcaggtcga cctcgaggac atgttctttc ctgcgttatc ccctgattct gtggataacc
      181 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      241 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
      301 gcggtatttc acaccgcata gggtcatggc tgcgccccga cacccgccaa cacccgctga
      361 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
      421 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagaagga
      481 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
      541 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
      601 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
      661 ctgctcatgt ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag
      721 ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt
      781 atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg
      841 ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa
      901 cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc
      961 tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat
     1021 gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc
     1081 tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg
     1141 gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta
     1201 tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca
     1261 ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata
     1321 ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt
     1381 gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca
     1441 gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac
     1501 cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat
     1561 cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt
     1621 atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca
     1681 gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc
     1741 tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga
     1801 ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca
     1861 ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag
     1921 cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttt
     1981 gggctagccg aagggtgaat tatgcatcac catcaccacc atcaccatca gcatcaacac
     2041 gaaaatctgt attttcagat ggctcttaac acaccacaaa ttacgccgac aaaaaagata
     2101 acagtgaggg caatcggcga ggaactgccg cgtggtgatt accaacgttg cccgcaatgt
     2161 gacatgctgt ttagcctgcc cgagataaat tctcatcaaa gtgcctattg tccgcgctgt
     2221 caggcaaaaa ttcgtgacgg gcgcgactgg tcgctaacgc gcctggcggc aatggccttc
     2281 accatgctgt tgcttatgcc gtttgcctgg ggcgaaccgc tgttgcatat ctggctgtta
     2341 ggcattcgta ttgacgccaa cgttatgcaa ggcatctggc aaatgaccaa acagggcgat
     2401 gcgataacgg ggtcgatggt ctttttctgc gttattggtg ccccgctcat tctggtgacc
     2461 tccatagctt atttatggtt tggtaaccga ctgggaatga atttacgtcc ggtactgctg
     2521 atgcttgagc gacttaaaga gtgggtaatg ctcgatatct acctggtcgg cattggcgtt
     2581 gcttctataa aggtacagga ttatgcccat atccaggctg gtgtcggctt gttctctttt
     2641 gtggcgttgg tgattttaac gacggtgacg ttgtcacatc ttaatgtcga agaactgtgg
     2701 gagcgatttt atccgcagcg ccccgctacg cgtagggacg agaaacttcg tgtctgtctt
     2761 gggtgccatt ttaccggcta tccagatcag cgtggtcgct gcccgcgttg ccatatcccg
     2821 ctacgcctgc gtcgccgtca tagtctgcaa aaatgctggg cggcgctgtt agcgtcaatc
     2881 gttttattgt tacctgccaa cctgttgcct atttctatca tttatctgaa cggaggacgg
     2941 caggaagata caattctttc cggaattatg tcgctggcaa gtagcaacat tgcggttgca
     3001 ggaatcgtgt ttatcgccag tattctggta ccgtttacta aagtgatcgt catgttcact
     3061 ttactgttga gcattcattt taaatgccag caaggtttac gcacacgcat tctgttactg
     3121 cggatggtga cctggattgg tcgctggtcg atgctcgacc tgtttgtcat atctttaacc
     3181 atgtcgctga ttaatcgcga tcagatcctc gcttttacta tgggaccggc tgcgttttat
     3241 ttcggcgcag cggtaatttt gactattctt gctgtggaat ggctggacag ccgcttactt
     3301 tgggatgcac atgagtcagg aaacgcccgc ttcgacgact gaagcgcaga ttaaaaataa
     3361 acgccgtatc tcacctttct ggctgctgcc gttcatcgcg ctaatgattg ccagttggct
     3421 gatttgggac agttatcagg accggggtaa taccgtcacc atcgacttta tgtcggcgga
     3481 tggtattgtt ccaggccgta cgcctgttcg ttatcagggc gttgaagtcg gaacagtgca
     3541 ggatatcagc ctcagcgacg atcttcgtaa gattgaagtc aaggtcagca tcaagtccga
     3601 tatgaaagat gcgctgcgcg aagagactca gttctggctg gtgacgccaa aagcatcgtt
     3661 ggcaggtgtc tccgggctgg acgccctcgt cggtgggaac tatatcggca tgatgccggg
     3721 taaaggtaaa gagcaggatc actttgtcgc actcgatacc caaccgaaat atcggctgga
     3781 caatggcgat ctgatgatcc acctgcaagc ccccgatctc ggttcgctga acagcggttc
     3841 attggtctat ttccgcaaga tcccggtggg aaaagtctac gactatgcca tcaatcccaa
     3901 caagcaaggc gtggtgattg atgtcctgat cgagcggcgt tttaccgatc tggtgaaaaa
     3961 aggtagccgt ttctggaacg tttccggcgt tgatgccaac gtcagtatca gtggcgcgaa
     4021 ggtgaaactg gaaagcctgg cggcactggt taacggtgcg attgccttcg attcaccaga
     4081 agagtcgaaa cctgccgagg cggaagatac ctttggtctg tatgaagatc tggcccacag
     4141 ccagcgtggc gtaataataa aactggaact gccgagtggg gccggattaa ccgccgactc
     4201 gacgccgtta atgtatcagg ggctggaagt cggacagctg actaaactgg atttaaatcc
     4261 tggtggtaaa gtcacagggg aaatgaccgt tgatcccagc gtcgttaccc tgcttcgtga
     4321 aaatacccgc atcgaattac gcaacccgaa attatccctt agcgatgcta atctcagcgc
     4381 cctgctgacc ggcaaaacct tcgagctggt gcccggcgat ggcgagccac gcaaagagtt
     4441 cgttgttgtg ccaggcgaaa aagcactgct gcatgaacct gatgttctga cgctgaccct
     4501 gaccgcgccg gaaagttacg gtattgatgc gggtcagccg ctcattcttc acggcgtgca
     4561 ggtaggccag gttatcgatc gtaaactcac cagcaaaggc gtcaccttta ccgtcgccat
     4621 cgagcctcag catcgcgaac tggtaaaagg cgatagcaaa tttgtcgtca acagccgtgt
     4681 cgatgtgaag gtggggctgg atggcgttga gtttctcggt gccagcgcct cagaatggat
     4741 caatggcggg atacgtattc tgccgggcga taaaggtgag atgaaagcca gctatccact
     4801 gtatgccaat ttggaaaaag cgctggagaa cagccttagc gatttaccca ccacaaccgt
     4861 gagtttgagt gcagagacgc tgccggatgt gcaggcagga tcggtagtgc tgtaccgtaa
     4921 atttgaagtt ggtgaagtga ttaccgtgcg tccgcgagct aacgcgtttg atatcgatct
     4981 gcatattaag ccggagtatc gcaaccttct gaccagcaat agcgtgttct gggcagaagg
     5041 cggggcgaaa gttcagctga atggtagtgg tctgaccgta caggcatccc cgctctccag
     5101 agcattaaag ggagccatta gcttcgacaa cctcagcggt gccagcgcca gtcagcgtaa
     5161 aggcgataag cgtattctgt atgcttccga aacagcggcc cgtgcggttg gcgggcagat
     5221 tacgcttcac gctttcgatg ccggaaaact ggcggtcggg atgccaattc gctatctcgg
     5281 tattgatatc gggcaaatcc agacgctgga tctgattacc gcacgcaatg aagtacaggc
     5341 aaaggcggta ctttacccgg aatatgtcca gacctttgct cgcggtggta cgcgcttctc
     5401 agtggtcacg ccaaaagcat cgttggcagg tgtctccggg ctggacgccc tcgtcggtgg
     5461 gaactatatc aacgtcgaac caggccgggg caatcctcgc cgcgactttg aattacaaga
     5521 ggccaccatt actgattcgc gttacctgga tggcttaagc attattgttg aagcgccgga
     5581 agccggttcg ttaggcatcg gtacgcctgt gctgttccgt ggtctggaag tcggtacggt
     5641 tacaggaatg acgctgggga cattgtcaga tcgcgtgatg attgcgatgc gcatcagtaa
     5701 acgctatcaa cacctggtgc gtaacaattc cgtcttctgg ttggcatcgg gttacagtct
     5761 ggactttggt ctgacgggcg gcgtagtgaa aaccggcacc tttaaccagt ttatccgtgg
     5821 cggcatcgcc ttcgccacgc ctccgggtac gccactggca ccgaaagccc aggaaggcaa
     5881 acacttcctg ttgcaggaaa gtgaaccgaa agagtggcgt gaatggggaa ctgcgcttcc
     5941 caaataagta ccaaattcca gaaaagagac gctgaaaagc gtcttttttc gttttggtcc
     6001 tggtctgagg aatggttgta agcttggctg ttttggcgga tgagagaaga ttttcagcct
     6061 gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag
     6121 tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga
     6181 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     6241 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     6301 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     6361 ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga
     6421 cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat
     6481 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6541 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6601 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     6661 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     6721 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     6781 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     6841 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6901 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6961 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7021 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7081 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7141 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7201 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7261 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7321 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7381 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7441 gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     7501 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     7561 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     7621 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     7681 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     7741 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga
     7801 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     7861 atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc taggtgaaga
     7921 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7981 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     8041 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     8101 taccaactct ttttccgaag gtaactggct cagcagagcg cagataccaa atactgtcct
     8161 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     8221 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     8281 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     8341 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     8401 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     8461 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     8521 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     8581 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     8641 ctggcctttt gctcacatgt tagc
//
