LOCUS       sequence-228304-        5884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5884)
  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 5884)
  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: pMSP1
FEATURES             Location/Qualifiers
     source          1..5884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(41..58)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(684..695)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(696..713)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             726..748
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    763..787
                     /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(787..806)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(788..806)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(890..909)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1115..1192
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1193..2275
                     /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(1212..1231)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3084..3275
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3269..3291)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3377..3516
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3432..3451
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3531..3548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3702..4290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3782..3801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4401..4420)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4412..5227
                     /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             4412..5227
                     /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(4485..4504)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5319..5774)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5456..5477)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5668..5687
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(5811..5858)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     5854..5872
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 actcagcttc ctttcgggct ttgttagcag ccggatctca gtggtggtgg tggtggtgct
       61 cgagtgcggc cgcaagctta ttactgggta ttcagctttt tagtatattc ttccagagct
      121 gacagaaaac tgactttaaa gctctcaaga acaggcaata agccctggcg tagatcttcc
      181 agcgccggtt tcgctttttc gctcaaggtg ctcaggtgtt ctgtcgcttt cgcgtggtac
      241 tcagccaagc gggcaccccc gttttctttc agcgcctcga gacgtgcggc caaacgctgg
      301 cgaagttcat ccgaatacgg cgccaaatga gtccggagtg catcaacatg tgcacgggcg
      361 cgatcgcgca tttcttcgcc taatgggctg agcttctctt ggagctcatg gagtttttgg
      421 cgtgccccct cttgcagttc cgcacgcagc ggttccacct tttgacgata taattccatc
      481 tcttcctgcc attttttctg aaagtcatct agatatggct gaaccttggc cttcacctct
      541 tctaaatctt tggacatttc ctgacgcagt ccctcggttt ctttttccag gttgtcccag
      601 aattcctgcg tcacggggcc cagttgttcg cgaagtttac tgaaggtaga cgtaacagag
      661 tcccaattgt ccaacagctt cagacgtccc tcaatgtgat gatgatgatg atgacccatg
      721 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
      781 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
      841 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
      901 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
      961 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     1021 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     1081 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
     1141 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     1201 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     1261 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     1321 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     1381 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     1441 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     1501 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     1561 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     1621 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     1681 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     1741 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     1801 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     1861 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     1921 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     1981 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     2041 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     2101 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     2161 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2221 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2281 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
     2341 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     2401 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     2461 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     2521 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     2581 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
     2641 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
     2701 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
     2761 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
     2821 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
     2881 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
     2941 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
     3001 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
     3061 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
     3121 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
     3181 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
     3241 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3301 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3361 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     3421 gcttaactat gcggcatcag agcagattgt actgagagtg caccattgcg gtgtgaaata
     3481 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     3541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4141 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4261 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     4321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc
     4381 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     4441 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     4501 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     4561 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     4621 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     4681 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     4741 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     4801 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     4861 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     4921 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     4981 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     5041 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     5101 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     5161 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     5221 tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5281 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5341 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5401 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5461 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5521 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5581 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5641 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5701 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5761 gctacagggc gcgtcccatt cgccaatccg gatatagttc ctcctttcag caaaaaaccc
     5821 ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc ggtggcagca
     5881 gcca
//
