LOCUS       sequence-212062-        4771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as N-terminal Sumo tag 
            fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4771)
  TITLE     Hyvonen Lab pOP plasmids
REFERENCE   2  (bases 1 to 4771)
  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: pOP3SU
FEATURES             Location/Qualifiers
     source          1..4771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    19..43
                     /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             56..78
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..136
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             146..169
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             176..469
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     promoter        complement(516..534)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(517..534)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(592..610)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      606..653
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(889..910)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1060..2142)
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2104..2123
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2143..2220)
                     /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     2366..2385
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2541..2560)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2660..2682
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2720..2738)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2806..2910
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2911..2979,2980..3771)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3942..4530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4431..4450
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4684..4701
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg aattgtgagc ggataacaat tcctctagaa ataattttgt
       61 ttaactttaa gaaggagata tatccatgaa tggactgaat gatatctttg aagcgcagaa
      121 aattgaatgg catgaatccg gatctcatca ccatcaccat caccatcaca ctagtatgtc
      181 ggactcagaa gtcaatcaag aagctaagcc agaggtcaag ccagaagtca agcctgagac
      241 tcacatcaat ttaaaggtgt ccgatggatc ttcagagatc ttcttcaaga tcaaaaagac
      301 cactccttta agaaggctga tggaagcgtt cgctaaaaga cagggtaagg aaatggactc
      361 cttaagattc ttgtacgacg gtattagaat tcaagctgat cagacccctg aagatttgga
      421 catggaggat aacgatatta ttgaggctca cagagaacag attggtggat ccatggaatt
      481 cgcggccgcc ctaggctcga gctaagcttg agtattctat agtgtcacct aaatcccagc
      541 ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      601 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
      661 gaggaactat atccggataa cctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
      721 ccaacagttg cgcagcctga atggcgaatg gaaattgtaa gcgttaatat tttgttaaaa
      781 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
      841 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
      901 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
      961 ggcgatggcc cacagcttgc atgcctgcag gtcggaagca taaagtgtaa agcctggggt
     1021 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1081 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1141 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     1201 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     1261 gcgaaaatcc tgtttgatgg tggttgacgg cgggatataa catgagctgt cttcggtatc
     1321 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     1381 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     1441 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     1501 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     1561 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     1621 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     1681 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     1741 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     1801 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     1861 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     1921 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     1981 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     2041 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     2101 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     2161 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt cgatggtgtc
     2221 gggtaccgag ctcgaattta cgtgaaccat cacccaaatc aagttttttg cggtcgaggt
     2281 gccgtaaagc tctaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2341 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2401 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2461 tacagggcgc gtcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2521 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     2581 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     2641 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     2701 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     2761 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2821 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2881 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2941 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3001 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3061 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3121 aaagttctgc tatgtcatac actattatcc cgtattgacg ccgggcaaga gcaactcggt
     3181 cgccgggcgc ggtattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3241 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3301 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3361 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3421 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3481 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3541 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3601 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3661 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3721 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3781 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3841 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3901 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3961 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4021 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4081 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4141 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4201 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4261 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4321 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4381 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4441 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4501 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4561 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4621 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4681 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4741 gcgcgttggc cgattcatta atgcagctga t
//
