LOCUS       sequence-257343-        6436 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression vector for the Escherichia coli MinD with an N-terminal 
            His-tag for purification. MinE is encoded in the ORF to avoid 
            toxicity when overexpressing MinD, but MinE is not tagged..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6436)
  AUTHORS   Loose M, Fischer-Friedrich E, Ries J, Kruse K, Schwille P
  TITLE     Spatial regulators for bacterial cell division self-organize into 
            surface waves in vitro.
  JOURNAL   Science. 2008 May 9;320(5877):789-92. doi: 10.1126/science.1154413.
  PUBMED    18467587
REFERENCE   2  (bases 1 to 6436)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2008-05-9- 9"; volume: "320"; issue: 
            "5877"; pages: "789-92"
COMMENT     Sequence Label: pET28a-His-MinD_MinE
FEATURES             Location/Qualifiers
     source          1..6436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1274..1306)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(1310..1327)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1337..1354)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1373..1395
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1410..1434
                     /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)."
     primer_bind     complement(1434..1453)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1435..1453)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1537..1556)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1762..1839
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1840..2922
                     /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     complement(1859..1878)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3731..3922
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3916..3938)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4024..4166
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4079..4098
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4181..4198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4352..4940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4432..4451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5051..5070)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5062..5877
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5062..5877
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5135..5154)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5970..6425)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6107..6128)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6319..6338
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagctttt
      181 atttcagctc ttctgcttcc ggtaaggtca cgttcagctc aagaatagaa atatcgccat
      241 ctttttgctc aagctgtacg gttaccatct caggatcaat ttgtacgtat ttacaaatga
      301 cctcaagaat atctttacgc aactgcggca gataatgcgg ttctgcatcg ctgcgacggc
      361 gttcagcaac aataatctgc agccgttctt ttgcaatgtt ggctgtgttt ttcttccgcg
      421 agagaaagaa atcgagtaat gccataactt atcctccgaa caagcgtttg aggaagcctt
      481 tcttctcttc ttcaatgaag cggaaaggac gttcttctcc caacagacgt tctacggtat
      541 ctgcgtaggc tttacccgca tcggcgttaa tgtcgagaat gaccggttca ccctggttag
      601 aggcgcgcaa tactgattga tcctctggga tcacgccgac gagtttgatg cgcaggatct
      661 ccagcacatc ttccatgctc agcatgtcac ctctgcttac gcggcctggg ttatagcgcg
      721 ttaacagcag gtgctcttta ataggctctt cgccattttc tgcgcggcgt gatttcgacg
      781 ccagaatgcc taaaatacgg tcagagtcgc gtactgagga gacttccggg ttggtggtaa
      841 taatggcttc gtctgcaaaa tagagtgcca ttaacgcacc ggtttcaatc cctgccgggg
      901 agtcacaaac gataaattca aaatccatcg ctttcagatc atcaagaact ttggcgaccc
      961 cttcacgggt gagggcatct ttatcgcgtg tttgcgatgc cggcagaata tagagatttt
     1021 cagtacgctt atctttaatt aacgcctgat ttagcgttgc atcgccctga atgacgttga
     1081 cgaaatcgta aacgacccgg cgttcacaac ccataatcag gtcgagatta cgcaggccga
     1141 tatcaaaatc tatcacgaca gttttctttc ccttctgggc caaaccagtg gcgatggccg
     1201 cgctggaggt tgtcttacca acaccccctt tgcccgaagt aacaacaata atgcgtgcga
     1261 attcggatcc gcgacccatt tgctgtccac cagtcatgct agccatatgg ctgccgcgcg
     1321 gcaccaggcc gctgctgtga tgatgatgat gatggctgct gcccatggta tatctccttc
     1381 ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata
     1441 gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg
     1501 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
     1561 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
     1621 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg
     1681 cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata
     1741 agggagagcg tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca
     1801 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
     1861 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
     1921 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
     1981 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
     2041 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
     2101 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
     2161 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
     2221 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
     2281 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
     2341 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
     2401 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
     2461 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
     2521 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
     2581 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
     2641 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
     2701 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
     2761 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
     2821 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2881 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2941 aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc
     3001 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     3061 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     3121 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
     3181 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     3241 cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt cgttgaggac
     3301 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg
     3361 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt
     3421 cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt
     3481 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
     3541 gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag
     3601 ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt
     3661 gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac
     3721 ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag
     3781 ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat
     3841 ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg
     3901 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3961 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4021 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     4081 gcatcagagc agattgtact gagagtgcac catatatgcg gtgtgaaata ccgcacagat
     4141 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     4201 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4981 ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata
     5041 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg
     5101 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     5161 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
     5221 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     5281 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     5341 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
     5401 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     5461 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     5521 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     5581 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     5641 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     5701 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     5761 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     5821 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagaa
     5881 ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5941 gcgcacattt ccccgaaaag tgccacctga aattgtaaac gttaatattt tgttaaaatt
     6001 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6061 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6121 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6181 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6241 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6301 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6361 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6421 cgcgtcccat tcgcca
//
