LOCUS       sequence-271463-        6432 bp    DNA     circular SYN 15-DEC-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 6432)
  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 6432)
  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"; 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..6432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             148..180
                     /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             1297..1314
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1367..1385)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1381..1428
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1465..1920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1552..1571)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1762..1783
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2012..2827)
                     /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             complement(2012..2827)
                     /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     2735..2754
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2819..2838
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2949..3537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3438..3457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3691..3708
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3723..3862
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3788..3807)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3948..3970
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3964..4155)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4964..6046)
                     /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     6008..6027
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6047..6124)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6330..6349
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg gctagcatga ctggtggaca gcaaatgggt
      181 cgcggatccg aattcgcacg cattattgtt gttacttcgg gcaaaggggg tgttggtaag
      241 acaacctcca gcgcggccat cgccactggt ttggcccaga agggaaagaa aactgtcgtg
      301 atagattttg atatcggcct gcgtaatctc gacctgatta tgggttgtga acgccgggtc
      361 gtttacgatt tcgtcaacgt cattcagggc gatgcaacgc taaatcaggc gttaattaaa
      421 gataagcgta ctgaaaatct ctatattctg ccggcatcgc aaacacgcga taaagatgcc
      481 ctcacccgtg aaggggtcgc caaagttctt gatgatctga aagcgatgga ttttgaattt
      541 atcgtttgtg actccccggc agggattgaa accggtgcgt taatggcact ctattttgca
      601 gacgaagcca ttattaccac caacccggaa gtctcctcag tacgcgactc tgaccgtatt
      661 ttaggcattc tggcgtcgaa atcacgccgc gcagaaaatg gcgaagagcc tattaaagag
      721 cacctgctgt taacgcgcta taacccaggc cgcgtaagca gaggtgacat gctgagcatg
      781 gaagatgtgc tggagatcct gcgcatcaaa ctcgtcggcg tgatcccaga ggatcaatca
      841 gtattgcgcg cctctaacca gggtgaaccg gtcattctcg acattaacgc cgatgcgggt
      901 aaagcctacg cagataccgt agaacgtctg ttgggagaag aacgtccttt ccgcttcatt
      961 gaagaagaga agaaaggctt cctcaaacgc ttgttcggag gataagttat ggcattactc
     1021 gatttctttc tctcgcggaa gaaaaacaca gccaacattg caaaagaacg gctgcagatt
     1081 attgttgctg aacgccgtcg cagcgatgca gaaccgcatt atctgccgca gttgcgtaaa
     1141 gatattcttg aggtcatttg taaatacgta caaattgatc ctgagatggt aaccgtacag
     1201 cttgagcaaa aagatggcga tatttctatt cttgagctga acgtgacctt accggaagca
     1261 gaagagctga aataaaagct tgcggccgca ctcgagcacc accaccacca ccactgagat
     1321 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     1381 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     1441 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1501 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1561 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1621 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1681 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1741 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1801 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1861 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     1921 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1981 ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa
     2041 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     2101 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     2161 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     2221 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     2281 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     2341 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     2401 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     2461 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     2521 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     2581 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     2641 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     2701 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     2761 aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat
     2821 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc
     2881 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2941 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3001 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3061 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3121 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3181 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3241 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3301 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3361 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3421 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3481 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3541 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3601 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3661 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3721 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     3781 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     3841 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     3901 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3961 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     4021 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     4081 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     4141 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     4201 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     4261 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     4321 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     4381 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     4441 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     4501 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     4561 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     4621 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
     4681 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
     4741 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
     4801 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
     4861 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
     4921 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
     4981 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5041 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
     5101 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
     5161 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
     5221 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
     5281 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
     5341 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
     5401 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
     5461 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
     5521 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
     5581 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
     5641 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
     5701 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
     5761 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
     5821 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
     5881 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
     5941 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
     6001 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
     6061 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
     6121 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     6181 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
     6241 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
     6301 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     6361 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     6421 atcccgcgaa at
//
