LOCUS       His-lsMSP1E3D1_p        6143 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 6143)
  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..6143
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        223..241
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    242..266
                     /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             297..302
                     /note="ribosome binding site"
     CDS             310..1206
                     /codon_start=1
                     /label=His-lsMSP1E3D1
                     /translation="MGHHHHHHHDYDIPTTENLYFQGSSFSKLREELGPVSEEFWDDLE
                     KESEGLREEMSKDLEEVKAKVEPYLDDFEKKWEEEMELYREKVEPLRAELEEGAREKLH
                     ELEEKLSPLGEEMRDRARAHVDALRSHLAPYLDDFEKKWEEEMELYREKVEPLRAELEE
                     GAREKLHELEEKLSPLGEEMRDRARAHVDALRSHLAPYSDELRERLAARLEALKEDGGA
                     RLAEYHAKASEHLSSLSEKAKPALEDLREGLLPVLESFKVSFLSALEEYSKKLDSEGGR
                     GGSLPETGGASHHHHHH"
     CDS             316..336
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             358..378
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1186..1203
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1213..1225
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1226..1243
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1310..1357
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1394..1849
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1942..2757)
                     /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      2879..3467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3897..4088)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4897..5979)
                     /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(5980..6057)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
       61 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
      121 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
      181 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
      241 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
      301 agatatacca tgggccacca ccatcatcat catcatgact acgatattcc gaccacggaa
      361 aacttatatt tccagggatc atcctttagc aagttgcgtg aggagctggg gccagtgtct
      421 gaagaatttt gggatgatct ggagaaggag agcgagggct tacgcgagga gatgtccaaa
      481 gatcttgagg aagtgaaggc taaagtggaa ccatatctgg acgactttga aaaaaaatgg
      541 gaagaagaga tggagttata tcgcgaaaaa gtggaaccct tgcgcgcgga acttgaggaa
      601 ggtgcgcgtg agaaacttca tgagttggaa gaaaagttat ctccgcttgg ggaagaaatg
      661 cgtgaccgtg ctcgtgcaca cgtcgatgca ctgcgctcgc acttggcccc atatttagat
      721 gatttcgaga aaaagtggga ggaggaaatg gaactttacc gcgagaaggt tgagcccttg
      781 cgcgcagaac ttgaggaggg tgcacgcgag aaacttcatg aacttgagga aaaactgtca
      841 cccttgggcg aggagatgcg tgatcgtgct cgcgctcatg tagacgcctt acgttcacac
      901 ttagcgccat actccgatga gttgcgcgag cgtcttgcag cacgtttgga agccttaaag
      961 gaagatggag gtgcacgttt ggcggaatat catgccaagg cgagcgagca cttgtccagc
     1021 ttgtcggaga aagccaaacc ggcactggaa gacctgcgcg aggggctgtt acctgtatta
     1081 gagagtttta aggtcagctt tttgtcagca cttgaggaat atagcaagaa attggatagt
     1141 gagggcggcc gcggtggcag cctgccggaa accggtggcg ctagccacca tcatcatcat
     1201 cattaagctt gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa
     1261 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1321 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1381 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1441 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1501 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1561 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1621 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1681 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     1741 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1801 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact
     1861 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1921 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1981 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     2041 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     2101 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     2161 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     2221 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     2281 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     2341 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     2401 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     2461 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     2521 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     2581 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     2641 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     2701 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     2761 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     2821 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2881 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2941 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3001 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3061 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3121 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3181 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3241 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3301 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3361 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3421 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3481 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3541 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3601 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     3661 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag
     3721 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3781 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3841 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3901 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3961 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     4021 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4081 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4141 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4201 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     4261 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     4321 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     4381 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4441 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4501 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     4561 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4621 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4681 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4741 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     4801 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     4861 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4921 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4981 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5041 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5101 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5161 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5221 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5281 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5341 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5401 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5461 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5521 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5581 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5641 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5701 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     5761 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     5821 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     5881 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     5941 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6001 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6061 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6121 ggccgttgag caccgccgcc gca
//
