LOCUS       sequence-489681-        5941 bp    DNA     circular SYN 17-OCT-2025
DEFINITION  Expression of the MSP 'His-lsMSP1D1'.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5941)
  AUTHORS   Tidemand FG, Blemmer S, Johansen NT, Arleth L, Pedersen MC
  TITLE     Non-ionic detergent assists formation of supercharged nanodiscs and 
            insertion of membrane proteins.
  JOURNAL   Biochim Biophys Acta Biomembr. 2022 Jun 1;1864(6):183884. doi: 
            10.1016/j.bbamem.2022.183884. Epub 2022 Feb 16.
  PUBMED    35182589
REFERENCE   2  (bases 1 to 5941)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biochim 
            Biophys Acta Biomembr."; date: "2022-06-1"; volume: "1864(6):183884.
            doi"; pages: " 10.1016/j.bbamem.2022.183884. Epub 2022 Feb 16"
COMMENT     Sequence Label: His-lsMSP1D1
FEATURES             Location/Qualifiers
     source          1..5941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     83..102
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        83..101
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    102..126
                     /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             141..163
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             176..196
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             218..238
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             848..865
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             888..905
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(958..976)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      972..1019
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1056..1511
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1143..1162)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1353..1374
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1603..2418)
                     /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(1603..2418)
                     /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     2326..2345
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2410..2429
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2540..3128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3029..3048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3282..3299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3314..3453
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3379..3398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3539..3561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3555..3746)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4555..5637)
                     /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     5599..5618
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5638..5715)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5921..5940
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
       61 cgagatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
      121 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca tgggccacca
      181 ccatcatcat catcatgact acgatattcc gaccacggaa aacttatatt tccagggatc
      241 atccttttca aaactgcgtg aagagcttgg ccctgtgagc gaggaattct gggacgatct
      301 ggagaaggag tctgaagggc tgcgcgaaga gatgagcaaa gacctggaag aagtgaaggc
      361 taaagtagag ccttatttgg acgatttcga gaaaaagtgg gaggaggaga tggagttgta
      421 tcgtgagaag gttgagcctc ttcgcgcaga gcttgaagaa ggcgctcgtg agaagttaca
      481 cgaacttgaa gaaaaattaa gcccgcttgg agaggaaatg cgtgatcgcg cgcgcgccca
      541 tgtagatgca cttcgtagcc atcttgcgcc ctacagcgat gagcttcgtg aacgcctggc
      601 agcccgttta gaagcgttga aggaagacgg tggtgcgcgc ctggcggagt atcatgcaaa
      661 ggcttcggaa catctttcct ctctttctga gaaagcgaaa cccgctctgg aagaccttcg
      721 cgaagggctt cttcctgtcc ttgaatcttt taaggtgagt tttttgtccg cattagaaga
      781 atactccaag aaattagatt ccgaaggcgg ccgcggtggc agcctgccgg aaaccggtgg
      841 cgctagccac catcatcatc atcattaagc ttgcggccgc actcgagcac caccaccacc
      901 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      961 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1021 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     1081 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1141 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1201 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1261 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1321 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1381 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1441 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1501 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1561 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     1621 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     1681 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     1741 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     1801 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     1861 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     1921 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     1981 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     2041 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     2101 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     2161 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     2221 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     2281 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     2341 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     2401 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     2461 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2521 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2581 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2641 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     2701 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2761 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2821 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2881 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2941 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3001 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3061 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3121 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3181 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3241 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3301 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3361 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3421 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     3481 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3541 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     3601 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     3661 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     3721 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     3781 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     3841 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     3901 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     3961 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     4021 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     4081 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     4141 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     4201 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     4261 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     4321 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     4381 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     4441 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     4501 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     4561 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4621 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     4681 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     4741 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     4801 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     4861 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4921 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     4981 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     5041 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     5101 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     5161 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     5221 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     5281 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     5341 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     5401 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     5461 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     5521 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     5581 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     5641 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     5701 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     5761 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     5821 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     5881 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     5941 c
//
