LOCUS       sequence-220203-        4412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as non-cleavable 
            C-terminal Strep Tag fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4412)
  TITLE     Hyvonen Lab pOP plasmids
REFERENCE   2  (bases 1 to 4412)
  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"
COMMENT     Sequence Label: pOP2S
FEATURES             Location/Qualifiers
     source          1..4412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /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             76..98
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             145..168
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(182..200)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(183..200)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(258..276)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      272..319
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(555..576)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(726..1808)
                     /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     1770..1789
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1809..1886)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     2029..2048
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2202..2221)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2321..2343
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2381..2399)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2467..2571
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2572..2640,2641..3432)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2790..2809)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3603..4191
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4092..4111
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4345..4362
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gattcattaa tgcagctgat taatacgact cactataggg aattgtgagc ggataacaat
       61 tcctctagaa ataattttgt ttaactttaa gaaggagata tatccatggg atccgaattc
      121 gcggccgccc taggctcgag cgcatggagc cacccgcagt tcgaaaaata agcttgagta
      181 ttctatagtg tcacctaaat cccagcttga tccggctgct aacaaagccc gaaaggaagc
      241 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
      301 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggataacctg gcgtaatagc
      361 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggaaa
      421 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
      481 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
      541 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
      601 tcaaagggcg aaaaaccgtc tatcagggcg atggcccaca gcttgcatgc ctgcaggtcg
      661 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaacttaca ttaattgcgt
      721 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
      781 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
      841 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
      901 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
      961 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     1021 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     1081 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     1141 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     1201 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     1261 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     1321 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     1381 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     1441 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     1501 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     1561 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     1621 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     1681 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     1741 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     1801 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     1861 aaggttttgc accattcgat ggtgtcgggt accgagctcg aattgtgaac catcacccta
     1921 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     1981 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2041 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2101 acccgccgcg cttaatgcgc cgctacaggg cgcgtcctga tgcggtattt tctccttacg
     2161 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     2221 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2281 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2341 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     2401 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     2461 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2521 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     2581 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2641 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2701 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2761 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2821 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2881 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2941 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3001 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3061 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3121 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3181 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3241 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3301 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3361 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3421 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3481 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     3541 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3601 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3661 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3721 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3781 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3841 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3901 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3961 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4021 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4081 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4141 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4201 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4261 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     4321 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     4381 atacgcaaac cgcctctccc cgcgcgttgg cc
//
