LOCUS       sequence-354445-        5881 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of membrane scaffold protein (MSP1D1) for 
            Nanodisc formation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5881)
  AUTHORS   Denisov IG, Grinkova YV, Lazarides AA, Sligar SG
  TITLE     Directed self-assembly of monodisperse phospholipid bilayer 
            Nanodiscs with controlled size.
  JOURNAL   J Am Chem Soc. 2004 Mar 24. 126(11):3477-87.
  PUBMED    15025475
REFERENCE   2  (bases 1 to 5881)
  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: "J Am Chem 
            Soc. 2004 Mar 24. 126(11):3477-87."
COMMENT     Sequence Label: pMSP1D1
FEATURES             Location/Qualifiers
     source          1..5881
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1366..1557
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1551..1573)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1659..1798
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1714..1733
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1813..1830)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1984..2572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2064..2083)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2683..2702)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2694..3509
                     /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             2694..3509
                     /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(2767..2786)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3601..4056)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3738..3759)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3950..3969
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4093..4140)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4136..4154
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4207..4224)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4814..4834)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(4856..4876)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     RBS             4889..4911
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4926..4950
                     /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(4950..4969)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4951..4969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5053..5072)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5278..5355
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5356..5881,1..557)
                     /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(5375..5394)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
       61 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
      121 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
      181 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
      241 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
      301 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
      361 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
      421 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
      481 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      541 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg
      601 ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
      661 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
      721 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
      781 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
      841 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggcccca
      901 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
      961 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     1021 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     1081 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     1141 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     1201 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     1261 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     1321 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     1381 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     1441 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     1501 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     1561 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     1621 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     1681 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     1741 tgcaccattg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     1801 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1861 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1921 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1981 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2041 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2101 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2161 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2221 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2281 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2341 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2401 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     2461 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2521 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     2581 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2641 ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     2701 atattcaacg ggaaacgtct tgctctaggc cgcgattaaa ttccaacatg gatgctgatt
     2761 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
     2821 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     2881 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     2941 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     3001 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     3061 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     3121 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     3181 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3241 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3301 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3361 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     3421 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     3481 ttcatttgat gctcgatgag tttttctaag aattaattca tgagcggata catatttgaa
     3541 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3601 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3661 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3721 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3781 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3841 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3901 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3961 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4021 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccaatc cggatatagt
     4081 tcctcctttc agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag
     4141 ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt tagcagccgg
     4201 atctcagtgg tggtggtggt ggtgctcgag tgcggccgca agcttactgg gtattcagct
     4261 ttttagtata ttcttccaga gctgacagaa aactgacttt aaagctctca agaacaggca
     4321 ataagccctg gcgtagatct tccagcgccg gtttcgcttt ttcgctcaag gtgctcaggt
     4381 gttctgtcgc tttcgcgtgg tactcagcca agcgggcacc cccgttttct ttcagcgcct
     4441 cgagacgtgc ggccaaacgc tggcgaagtt catccgaata cggcgccaaa tgagtccgga
     4501 gtgcatcaac atgtgcacgg gcgcgatcgc gcatttcttc gcctaatggg ctgagcttct
     4561 cttggagctc atggagtttt tggcgtgccc cctcttgcag ttccgcacgc agcggttcca
     4621 ccttttgacg atataattcc atctcttcct gccatttttt ctgaaagtca tcgagatatg
     4681 gctgaacctt ggccttcacc tcttctaaat ctttggacat ttcctgacgc agtccctcgg
     4741 tttctttttc caggttgtcc cagaattcct gcgtcacggg gcccagttgt tcgcgaagtt
     4801 tactgaaggt agaaccctga aaatacaaat tctcagtagt aggaatatca taatcgtgat
     4861 gatgatgatg atgatgaccc atggtatatc tccttcttaa agttaaacaa aattatttct
     4921 agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga
     4981 tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     5041 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     5101 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     5161 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
     5221 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac
     5281 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     5341 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     5401 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     5461 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     5521 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     5581 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     5641 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     5701 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     5761 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     5821 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     5881 g
//
