LOCUS       sequence-333773-        6246 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 6246)
  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 6246)
  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..6246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      486..1074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     975..994
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1228..1245
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1260..1402
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1328..1347)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1488..1510
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1504..1695)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2504..3586)
                     /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"
     primer_bind     3548..3567
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3587..3664)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3870..3889
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3973..3992
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3973..3991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3992..4016
                     /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             4031..4053
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4928..4957
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(5016..5034)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5030..5077
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5114..5569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5201..5220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5411..5432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5596..5700
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5701..5769,5770..6246,1..315)
                     /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(5919..5938)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
       61 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
      121 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
      181 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
      241 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
      301 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
      361 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
      421 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
      481 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
      541 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
      601 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
      661 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      721 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      781 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      841 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      901 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      961 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     1021 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     1081 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     1141 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     1201 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     1261 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca
     1321 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     1381 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     1441 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     1501 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     1561 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     1621 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     1681 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     1741 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     1801 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     1861 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     1921 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     1981 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     2041 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     2101 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     2161 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     2221 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     2281 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     2341 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     2401 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     2461 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     2521 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2581 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     2641 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     2701 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     2761 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     2821 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     2881 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     2941 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     3001 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     3061 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     3121 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     3181 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     3241 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     3301 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     3361 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     3421 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     3481 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     3541 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     3601 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     3661 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     3721 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     3781 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     3841 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     3901 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     3961 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
     4021 tagaaataat tttgtttaac tttaagaagg agatatacat atgaaaacct tcttcaaaac
     4081 cctttccgcc gccgcactcg cgctcatcct cgccgcctgc ggcggtcaaa aagatagcgc
     4141 gcccgccgca tccgcttctg ccgccgccga caacggcgcg gcgaaaaaag aaatcgtctt
     4201 cggcacgacc gtcggcgact tcggcgatat ggtcaaagaa caaatccaag ccgagctgga
     4261 gaaaaaaggc tacaccgtca aactggtcga gtttaccgac tatgtgcgcc cgaatctggc
     4321 attggctgag ggcgagttgg acatcaacgt cttccaacac aaaccctatc ttgacgactt
     4381 caaaaaagaa cacaatctgg acatcaccga agtcttccaa gtgccgaccg cgcctttggg
     4441 actgtacccg ggcaagctga aatcgctgga agaagtcaaa gacggcagca ccgtatccgc
     4501 gcccaacgac ccgtccaact tcgcccgcgt cttggtgatg ctcgacgaac tgggttggat
     4561 caaactcaaa gacggcatca atccgctgac cgcatccaaa gcggacattg ccgaaaacct
     4621 gaaaaacatc aaaatcgtcg agcttgaagc cgcgcaactg ccgcgtagcc gcgccgacgt
     4681 ggattttgcc gtcgtcaacg gcaactacgc cataagcagc ggcatgaagc tgaccgaagc
     4741 cctgttccaa gaaccgagct ttgcctatgt caactggtct gccgtcaaaa ccgccgacaa
     4801 agacagccaa tggcttaaag acgtaaccga ggcctataac tccgacgcgt tcaaagccta
     4861 cgcgcacaaa cgcttcgagg gctacaaatc ccctgccgca tggaatgaag gcgcagccaa
     4921 actcgagcat catcatcatc atcatcatca tcatcactga ctcgaggatc cggctgctaa
     4981 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     5041 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     5101 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5161 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5221 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5281 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5341 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5401 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5461 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5521 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact
     5581 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5641 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5701 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5761 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5821 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5881 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5941 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6001 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6061 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6121 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6181 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6241 cctgca
//
