LOCUS       sequence-344824-        6242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Methionine Binding Protein (MetQ) with signal 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6242)
  AUTHORS   Sharaf NG, Shahgholi M, Kim E, Lai JY, VanderVelde DG, Lee AT, Rees 
            DC
  TITLE     Characterization of the ABC methionine transporter from Neisseria 
            meningitidis reveals that lipidated MetQ is required for 
            interaction.
  JOURNAL   Elife. 2021 Aug 19;10. pii: 69742. doi: 10.7554/eLife.69742.
  PUBMED    34409939
REFERENCE   2  (bases 1 to 6242)
  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: "Elife."; 
            date: "2021-08-19"; volume: "10. pii: 69742. doi"; pages: " 
            10.7554/eLife.69742"
COMMENT     Sequence Label: NmMetQ
FEATURES             Location/Qualifiers
     source          1..6242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     939..958
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(978..1055)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1261..1280
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1364..1383
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1364..1382
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1383..1407
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1422..1444
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2319..2348
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(2407..2425)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2421..2468
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2505..2960
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2592..2611)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2802..2823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2986..3090
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3091..3159,3160..3951)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3309..3328)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4122..4710
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4611..4630
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4864..4881
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4896..5035
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4961..4980)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5121..5143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5137..5328)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6137..6242,1..977))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
       61 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
      121 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc
      181 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
      241 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
      301 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
      361 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
      421 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
      481 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
      541 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
      601 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
      661 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
      721 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
      781 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
      841 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
      901 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
      961 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
     1021 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
     1081 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     1141 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     1201 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     1261 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     1321 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
     1381 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
     1441 gagatataca tatgaaaacc ttcttcaaaa ccctttccgc cgccgcactc gcgctcatcc
     1501 tcgccgcctg cggcggtcaa aaagatagcg cgcccgccgc atccgcttct gccgccgccg
     1561 acaacggcgc ggcgaaaaaa gaaatcgtct tcggcacgac cgtcggcgac ttcggcgata
     1621 tggtcaaaga acaaatccaa cccgagctgg agaaaaaagg ctacaccgtc aaactggtcg
     1681 agtttaccga ctatgtgcgc ccgaatctgg cattggctga gggcgagttg gacatcaacg
     1741 tcttccaaca caaaccctat cttgacgact tcaaaaaaga acacaatctg gacatcaccg
     1801 aagtcttcca agtgccgacc gcgcctttgg gactgtaccc gggcaagctg aaatcgctgg
     1861 aagaagtcaa agacggcagc accgtatccg cgcccaacga cccgtccaac ttcgcccgcg
     1921 tcttggtgat gctcgacgaa ctgggttgga tcaaactcaa agacggcatc aatccgctga
     1981 ccgcatccaa agcggacatt gccgaaaacc tgaaaaacat caaaatcgtc gagcttgaag
     2041 ccgcgcaact gccgcgtagc cgcgccgacg tggattttgc cgtcgtcaac ggcaactacg
     2101 ccataagcag cggcatgaag ctgaccgaag ccctgttcca agaaccgagc tttgcctatg
     2161 tcaactggtc tgccgtcaaa accgccgaca aagacagcca atggcttaaa gacgtaaccg
     2221 aggcctataa ctccgacgcg ttcaaagcct acgcgcacaa acgcttcgag ggctacaaat
     2281 cccctgccgc atggaatgaa ggcgcagcca aactcgagca tcatcatcat catcatcatc
     2341 atcatcactg actcgaggat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     2401 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2461 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     2521 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2581 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2641 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2701 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2761 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2821 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2881 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2941 aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3001 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3061 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3121 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3181 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3241 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3301 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3361 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3421 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3481 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3541 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3601 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     3661 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3721 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3781 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3841 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3901 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3961 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4021 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4081 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4141 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4201 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4261 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4321 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4381 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4441 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4501 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4561 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4621 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4681 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4741 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4801 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4861 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4921 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     4981 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     5041 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5101 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5161 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     5221 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     5281 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     5341 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     5401 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     5461 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     5521 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     5581 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     5641 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     5701 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     5761 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     5821 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     5881 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     5941 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     6001 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     6061 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     6121 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6181 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     6241 tt
//
