LOCUS       sequence-270822-        6126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for mCherry fused to a tandem repeat of the E. 
            coli MinD membrane targeting sequence (MTS) (C-terminal fusion) and 
            an N-terminal His-tag for purification..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6126)
  AUTHORS   Ramm B, Glock P, Mucksch J, Blumhardt P, Garcia-Soriano DA, Heymann 
            M, Schwille P
  TITLE     The MinDE system is a generic spatial cue for membrane protein 
            distribution in vitro.
  JOURNAL   Nat Commun. 2018 Sep 26;9(1):3942. doi: 10.1038/s41467-018-06310-1.
  PUBMED    30258191
REFERENCE   2  (bases 1 to 6126)
  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: "Nat 
            Commun."; date: "2018-09-26"; volume: "9(1):3942. doi"; pages: " 
            10.1038/s41467-018-06310-1"
COMMENT     Sequence Label: pET28a-mCherry-2xEcMTS
FEATURES             Location/Qualifiers
     source          1..6126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     26..45
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    45..69
                     /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             84..106
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             125..142
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             176..883
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(321..339)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     589..608
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1016..1033
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1086..1104)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1100..1147
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1184..1639
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1481..1502
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1731..2546)
                     /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(1731..2546)
                     /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     2454..2473
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2538..2557
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2668..3256
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3157..3176
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3410..3427
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3442..3581
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3507..3526)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3667..3689
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3683..3874)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4683..5765)
                     /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     5727..5746
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5766..5843)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6049..6068
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
       61 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata ccatgggcag
      121 cagccatcat catcatcatc acggtggatc tggaggttca ggtggaagtc ctaggatggt
      181 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
      241 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
      301 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
      361 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
      421 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
      481 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
      541 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
      601 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
      661 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
      721 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
      781 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
      841 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagggatctg gcattgaaga
      901 agagaagaaa ggcttcctca aacgcttgtt cggaggaggc ggatctattg aagaagagaa
      961 gaaaggcttc ctcaaacgct tgttcggagg ataaaagctt gcggccgcac tcgagcacca
     1021 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1081 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1141 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     1201 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     1261 ctagcgcccg ctcctttcga tttcttccct tcctttctcg ccacgttcgc cggctttccc
     1321 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     1381 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     1441 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     1501 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     1561 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     1621 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1681 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     1741 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     1801 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     1861 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     1921 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     1981 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     2041 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     2101 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     2161 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     2221 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     2281 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     2341 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     2401 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     2461 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     2521 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     2581 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3241 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3301 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3361 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3421 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3481 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3541 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3601 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3661 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3721 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3781 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3841 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     3901 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     3961 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     4021 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     4081 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     4141 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     4201 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     4261 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     4321 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     4381 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     4441 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     4501 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     4561 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     4621 agggcatcgg tcgagatcct ggtgcctaat gagtgagcta acttacatta attgcgttgc
     4681 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4741 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     4801 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     4861 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     4921 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     4981 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     5041 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     5101 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     5161 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     5221 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     5281 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     5341 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     5401 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     5461 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     5521 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     5581 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     5641 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     5701 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     5761 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     5821 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     5881 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     5941 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     6001 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     6061 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     6121 gtagag
//
