LOCUS       Exported                6218 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses the mature metQ gene from Neisseria meningitides, without 
            the signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    NmMetQ WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   Nguyen PT, Lai JY, Kaiser JT, Rees DC
  TITLE     Structures of the Neisseria meningitides methionine-binding protein 
            MetQ in substrate-free form and bound to l- and d-methionine 
            isomers.
  JOURNAL   Protein Sci. 2019 Jul 26. doi: 10.1002/pro.3694.
  PUBMED    31348565
REFERENCE   2  (bases 1 to 6218)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6218
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(16..35)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     176..198
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(192..383)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1192..2274)
                     /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     2236..2255
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2275..2352)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2558..2577
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2661..2680
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2661..2679
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2680..2704
                     /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             2719..2741
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     2749..2814
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
     CDS             2821..2850
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2866..2880
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(3680..3698)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3694..3741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3778..4233
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3865..3884)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4075..4096
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4259..4363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4364..5224
                     /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(4582..4601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5395..5983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5884..5903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6137..6154
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6169..90
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
       61 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
      121 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
      181 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
      241 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
      301 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
      361 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
      421 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
      481 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
      541 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
      601 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
      661 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
      721 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
      781 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
      841 cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg
      901 gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac
      961 aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct
     1021 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     1081 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     1141 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     1201 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1261 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     1321 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     1381 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     1441 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     1501 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     1561 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     1621 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     1681 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     1741 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     1801 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     1861 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     1921 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     1981 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     2041 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     2101 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     2161 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     2221 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     2281 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     2341 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     2401 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     2461 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     2521 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     2581 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     2641 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     2701 ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat gaaatacctg
     2761 ctgccgaccg ctgctgctgg tctgctgctc ctcgctgccc agccggcgat ggccatgggc
     2821 catcatcatc atcatcatca tcatcatcac agcagcggcc atatcgacga cgacgacaag
     2881 catatgaaag aaatcgtctt cggcacgacc gtcggcgact tcggcgatat ggtcaaagaa
     2941 caaatccaag ccgagctgga gaaaaagggc tacaccgtca aactggtcga gtttaccgac
     3001 tatgtacgcc cgaatctggc attggctgag ggcgagttgg acatcaacgt cttccaacac
     3061 aaaccctatc ttgacgactt caaaaaagaa cacaatctgg acatcaccga agtcttccaa
     3121 gtgccgaccg cgcctttggg actgtacccg ggcaagctga aatcgctgga agaagtcaaa
     3181 gacggcagca ccgtatccgc gcccaacgac ccgtccaact tcgcccgcgt cttggtgatg
     3241 ctcgacgaac tgggttggat caaactcaaa gacggcatca atccgttgac cgcatccaaa
     3301 gcggacatcg ccgagaacct gaaaaacatc aaaatcgtcg agcttgaagc cgcgcaactg
     3361 ccgcgtagcc gcgccgacgt ggattttgcc gtcgtcaacg gcaactacgc cataagcagc
     3421 ggcatgaagc tgaccgaagc cctgttccaa gaaccgagct ttgcctatgt caactggtct
     3481 gccgtcaaaa ccgccgacaa agacagccaa tggcttaaag acgtaaccga ggcctataac
     3541 tccgacgcgt tcaaagccta cgcgcacaaa cgcttcgagg gctacaaatc ccctgccgca
     3601 tggaatgaag gcgcagccaa ataactcgag gatccggctg ctaacaaagc ccgaaaggaa
     3661 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     3721 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     3781 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3841 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3901 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3961 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4021 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4081 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4141 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4201 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     4261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4321 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4381 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4441 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4501 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4561 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4621 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4681 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4741 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4801 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4861 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     4921 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4981 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5041 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5101 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5161 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5221 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5521 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     5581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6061 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     6121 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     6181 ttttctcctt acgcatctgt gcggtatttc acaccgca
//
