LOCUS       sequence-257344-        5509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for the Escherichia coli MinE with an C-terminal 
            fusion of a His-tag for purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5509)
  AUTHORS   Glock P, Ramm B, Heermann T, Kretschmer S, Schweizer J, Mucksch J, 
            Alagoz G, Schwille P
  TITLE     Stationary patterns in a two-protein reaction-diffusion system.
  JOURNAL   ACS Synth Biol. 2018 Dec 20. doi: 10.1021/acssynbio.8b00415.
  PUBMED    30571913
REFERENCE   2  (bases 1 to 5509)
  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: "ACS Synth 
            Biol. 2018 Dec 20. doi: 10.1021/acssynbio.8b00415."
COMMENT     Sequence Label: pET28a-MinE-His
FEATURES             Location/Qualifiers
     source          1..5509
                     /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"
     RBS             446..468
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    483..507
                     /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(507..526)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(508..526)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(610..629)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        835..912
                     /gene="lacI"
                     /label=lacI promoter
     CDS             913..1995
                     /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(932..951)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2804..2995
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2989..3011)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3097..3239
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3152..3171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3254..3271)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3425..4013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3505..3524)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4124..4143)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4135..4950
                     /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             4135..4950
                     /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(4208..4227)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5043..5498)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5180..5201)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5392..5411
                     /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 gactccagat ccacctttca
      181 gctcttctgc ttccggtaag gtcacgttca gctcaagaat agaaatatcg ccatcttttt
      241 gctcaagctg tacggttacc atctcaggat caatttgtac gtatttacaa atgacctcaa
      301 gaatatcttt acgcaactgc ggcagataat gcggttctgc atcgctgcga cggcgttcag
      361 caacaataat ctgcagccgt tcttttgcaa tgttggctgt gtttttcttc cgcgagagaa
      421 agaaatcgag taatgccatg gtatatctcc ttcttaaagt taaacaaaat tatttctaga
      481 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
      541 agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
      601 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
      661 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
      721 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
      781 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
      841 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
      901 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
      961 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     1021 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     1081 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     1141 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     1201 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     1261 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     1321 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     1381 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     1441 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     1501 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     1561 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     1621 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     1681 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     1741 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     1801 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     1861 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     1921 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     1981 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
     2041 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     2101 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     2161 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     2221 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     2281 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg
     2341 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     2401 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     2461 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     2521 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     2581 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     2641 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     2701 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     2761 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     2821 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     2881 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     2941 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     3001 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     3061 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     3121 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     3181 caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     3241 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3301 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3361 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3421 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3481 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3541 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3601 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3661 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3721 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3781 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3841 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     3901 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3961 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4021 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4081 tggtcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     4141 catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat
     4201 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
     4261 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     4321 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     4381 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     4441 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     4501 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     4561 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     4621 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     4681 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     4741 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     4801 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     4861 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     4921 tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga
     4981 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5041 tgaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5101 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5161 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5221 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5281 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5341 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5401 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5461 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca
//
