LOCUS       His-lsMSP1D1dH5_        5885 bp    DNA     circular SYN 16-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5885)
  AUTHORS   SnapGene License
  TITLE     Direct Submission
  JOURNAL   Exported Jun 16, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5885
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        100..118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    119..143
                     /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             174..179
                     /note="ribosome binding site"
     CDS             187..825
                     /codon_start=1
                     /label=His-lsMSP1D1dH5
                     /translation="MGHHHHHHHDYDIPTTENLYFQGSSSSFSKLREELGPVSEEFWDD
                     LEKESEGLREEMSKDLEEVKAKVEPYLDDFEKKWEEEMELYREKVEPLGEEMRDRARAH
                     VDALRSHLAPYSDELRERLAARLEALKEDGGARLAEYHAKASEHLSSLSEKAKPALEDL
                     REGLLPVLESFKVSFLSALEEYSKKLDSEGGRGGSLPETGGASHHHHHH"
     CDS             193..213
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             235..255
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             805..822
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    829..844
                     /label=MCS
                     /note="multiple cloning site"
     CDS             845..862
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      929..976
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1013..1468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1561..2376)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2498..3086
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3516..3707)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4516..5598)
                     /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"
     promoter        complement(5599..5676)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
       61 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg
      121 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
      181 tataccatgg gccaccacca tcatcatcat catgactacg atattccgac cacggaaaac
      241 ttatatttcc agggatcatc ctcgtccttc agcaaacttc gtgaggaact tggtccggtg
      301 agcgaggagt tctgggacga tctggaaaaa gaatccgaag gactgcgcga ggaaatgagt
      361 aaggatcttg aagaagtcaa agcaaaggtg gaaccatatc tggacgattt cgagaagaaa
      421 tgggaggagg agatggaact ttatcgtgag aaagttgaac ctttaggaga ggaaatgcgc
      481 gaccgtgccc gcgcacacgt cgatgcgctg cgctcccacc ttgctcccta ctccgatgaa
      541 cttcgtgagc gcttggctgc ccgtttggag gcgttaaaag aagatggcgg tgcccgcctg
      601 gccgaatatc acgccaaagc atcggagcat cttagctcgt tatcagaaaa ggcgaagcca
      661 gcccttgagg atctgcgtga gggtttactt cctgtgttgg aaagttttaa ggttagcttt
      721 ttatcggcgt tagaggagta tagcaagaaa ctggattctg agggcggccg cggtggcagc
      781 ctgccggaaa ccggtggcgc tagccaccat catcatcatc attaagcttg cggccgcact
      841 cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
      901 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
      961 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
     1021 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1081 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1141 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1201 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1261 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1321 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1381 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1441 tttaacaaaa tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga
     1501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
     1561 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
     1621 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     1681 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     1741 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     1801 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
     1861 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     1921 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     1981 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     2041 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     2101 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     2161 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     2221 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     2281 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     2341 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     2401 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     2461 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2521 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2581 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2641 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2701 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2761 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2821 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2881 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2941 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3001 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3061 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3121 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3181 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3241 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3301 tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc
     3361 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     3421 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3481 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3541 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     3601 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     3661 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     3721 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     3781 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     3841 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     3901 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     3961 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4021 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4081 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4141 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     4201 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     4261 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     4321 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     4381 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     4441 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     4501 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4561 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     4621 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     4681 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     4741 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     4801 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     4861 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     4921 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     4981 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5041 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5101 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5161 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     5221 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     5281 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     5341 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     5401 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     5461 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     5521 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     5581 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     5641 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     5701 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     5761 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     5821 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     5881 cggtg
//
