LOCUS       His-lsMSP1D1_pET        5945 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 5945)
  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..5945
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     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             157..162
                     /note="ribosome binding site"
     CDS             170..868
                     /codon_start=1
                     /label=His-lsMSP1D1
                     /translation="MGHHHHHHHDYDIPTTENLYFQGSSFSKLREELGPVSEEFWDDLE
                     KESEGLREEMSKDLEEVKAKVEPYLDDFEKKWEEEMELYREKVEPLRAELEEGAREKLH
                     ELEEKLSPLGEEMRDRARAHVDALRSHLAPYSDELRERLAARLEALKEDGGARLAEYHA
                     KASEHLSSLSEKAKPALEDLREGLLPVLESFKVSFLSALEEYSKKLDSEGGRGGSLPET
                     GGASHHHHHH"
     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"
     misc_feature    872..887
                     /label=MCS
                     /note="multiple cloning site"
     CDS             888..905
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     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"
     CDS             complement(1604..2419)
                     /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      2541..3129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3559..3750)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4559..5641)
                     /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(5642..5719)
                     /gene="lacI"
                     /label=lacI promoter
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 gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1561 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     1621 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1681 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1741 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1801 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     1861 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     1921 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     1981 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     2041 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     2101 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     2161 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     2221 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     2281 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     2341 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     2401 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     2461 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2521 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2581 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2641 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2701 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2761 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2821 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2881 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2941 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3001 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3061 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3121 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3181 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3241 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3301 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3361 cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     3421 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     3481 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3541 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     3601 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     3661 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     3721 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     3781 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     3841 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     3901 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     3961 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     4021 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     4081 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     4141 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     4201 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     4261 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     4321 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     4381 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     4441 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4501 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4561 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4621 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4681 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4741 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4801 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     4861 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     4921 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     4981 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     5041 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     5101 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5161 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5221 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5281 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5341 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5401 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5461 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5521 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5581 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5641 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5701 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     5761 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     5821 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     5881 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     5941 taggc
//
