LOCUS       sequence-237561-        5576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of P22 virus like particle in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5576)
  TITLE     Trevor Douglas lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5576)
  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: 
            "Unpublished"
COMMENT     Sequence Label: P22_wtCP_SP141his
FEATURES             Location/Qualifiers
     source          1..5576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             51..68
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             80..97
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        687..705
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    706..730
                     /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)."
     CDS             2081..2125
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2163..2181)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2177..2224
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2454..3269)
                     /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(2454..3269)
                     /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     3177..3196
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3270..3361)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3377..4126)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3387..3404)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3627..3646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4322..5404)
                     /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     5366..5385
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5405..5482)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        5528..5546
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5547..5571
                     /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)."
ORIGIN
        1 agaaataatt ttgtttaact ttaataagga gatataccat gggcagcagc catcaccatc
       61 atcaccacag ccaggatccc tggtgccgcg cggcagcatg tcgcagcaat gccgtagcag
      121 aacagggccg caagactcag gagtttaccc agcaatcagc gcaatacgtc gaagctgccc
      181 gcaaacacta tgacgcggcg gaaaagctca acatccctga ctatcaggag aaagaagacg
      241 catttatgca actggttccg cctgcggttg gggccgacat tatgcgcctg ttcccggaaa
      301 agtccgccgc gctcatgtat cacctggggg caaacccgga gaaagcccgc cagttactgg
      361 cgatggatgg gcagtccgcg ctgattgaac tcactcgact atccgaacgc ttaactctca
      421 agcctcgcgg taaacaaatc tcttccgctc cccatgctga ccagcctatt accggtgatg
      481 tcagcgcagc aaataaagat gccattcgta aacaaatgga tgctgctgcg agcaagggag
      541 atgtggaaac ctaccgcaag ctaaaggcaa aacttaaagg aatccgataa gagctcggcg
      601 cgcctgcagg tcgacaagct tgcggccgca taatgcttaa gtcgaacaga aagtaatcgt
      661 attgtacacg gccgcataat cgaaattaat acgactcact ataggggaat tgtgagcgga
      721 taacaattcc ccatcttagt atattagtta agtataagaa ggagatatac atatggcttt
      781 gaacgaaggt caaattgtta cactggcggt agatgaaatc atcgaaacca tctccgcaat
      841 cactccaatg gcgcagaaag ccaagaaata caccccgcct gctgcttcta tgcagcgctc
      901 cagcaatacc atctggatgc ctgtagagca agagtcaccc actcaggagg gctgggattt
      961 aactgataaa gcgacagggt tactggaact taacgtcgcg gtaaacatgg gagagccgga
     1021 taacgacttc ttccagttgc gtgctgatga cttgcgagac gaaactgcgt atcgtcgccg
     1081 catccagtct gccgctcgca agctggcgaa caacgttgag ttgaaagtcg caaacatggc
     1141 cgccgagatg ggttcgctgg ttatcacctc ccctgatgcc atcggcacta ataccgcaga
     1201 cgcctggaac tttgtggccg acgcagaaga aatcatgttc tcccgcgaac ttaaccgcga
     1261 catggggaca tcgtacttct tcaaccctca ggactacaaa aaagcgggtt acgacctgac
     1321 caagcgtgac atcttcgggc gtattcctga agaagcatac cgagatggca ccattcagcg
     1381 tcaggtcgct ggcttcgatg atgtcctgcg ctctccgaaa cttcctgtgc tgaccaaatc
     1441 caccgcaact ggcatcactg tatccggtgc gcagtccttc aagcctgtcg catggcaact
     1501 ggataacgat ggcaacaaag ttaacgttga taaccgtttt gctaccgtca ccctgtctgc
     1561 aactaccggc atgaaacgcg gcgacaaaat ttcgtttgct ggcgttaagt tccttggtca
     1621 gatggctaag aacgtactgg ctcaggatgc gactttctcc gtagtccgcg ttgttgacgg
     1681 tactcatgtt gaaatcacgc cgaagccggt agcgctggat gatgtttccc tgtctccgga
     1741 gcagcgtgcc tacgccaacg ttaacacctc gctggctgat gcaatggcag tgaacattct
     1801 gaacgttaaa gacgctcgca ctaatgtgtt ctgggctgac gatgctattc gtatcgtgtc
     1861 tcagccgatt ccggctaacc atgaactttt tgcaggtatg aaaactacct cattcagcat
     1921 ccctgatgtt ggcctgaacg gtatcttcgc tacgcagggt gatatttcca ccctgtccgg
     1981 cctgtgccgt attgcgctgt ggtacggcgt aaacgcgaca cgaccggagg caatcggtgt
     2041 tggcctgcct ggtcagactg cgtaatagct cgagtctggt aaagaaaccg ctgctgcgaa
     2101 atttgaacgc cagcacatgg actcgtctac tagcgcagct taattaacct aggctgctgc
     2161 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     2221 tttgctgaaa cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa
     2281 accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac
     2341 aagctgacga ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta
     2401 tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa
     2461 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     2521 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2581 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2641 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2701 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     2761 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2821 gcgagacgaa atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2881 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     2941 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     3001 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     3061 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     3121 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     3181 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag
     3241 caagacgttt cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt
     3301 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3361 ataggcatgc agcgctcttc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga
     3421 ctgcggcgag cggtgtcagc tcactcaaaa gcggtaatac ggttatccac agaatcaggg
     3481 gataaagccg gaaagaacat gtgagcaaaa agcaaagcac cggaagaagc caacgccgca
     3541 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagcca
     3601 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3661 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3721 gggaagcgtg gcgctttctc atagctcacg ctgttggtat ctcagttcgg tgtaggtcgt
     3781 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3841 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3901 cattggtaac tgatttagag gactttgtct tgaagttatg cacctgttaa ggctaaactg
     3961 aaagaacaga ttttggtgag tgcggtcctc caacccactt accttggttc aaagagttgg
     4021 tagctcagcg aaccttgaga aaaccaccgt tggtagcggt ggtttttctt tatttatgag
     4081 atgatgaatc aatcggtcta tcaagtcaac gaacagctat tccgttactc tagatttcag
     4141 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     4201 tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     4261 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     4321 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4381 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     4441 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     4501 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     4561 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     4621 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     4681 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     4741 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     4801 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     4861 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     4921 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     4981 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     5041 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     5101 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     5161 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     5221 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     5281 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     5341 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     5401 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     5461 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
     5521 aggaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctgt
//
