LOCUS       sequence-212060-        4390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as non-fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4390)
  TITLE     Hyvonen Lab pOP plasmids
REFERENCE   2  (bases 1 to 4390)
  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: pOP1
FEATURES             Location/Qualifiers
     source          1..4390
                     /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)"
     promoter        complement(135..153)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(136..153)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(211..229)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      225..272
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(508..529)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(679..1761)
                     /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     1723..1742
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1762..1839)
                     /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     1985..2004
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2160..2179)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2279..2301
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2339..2357)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2425..2529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2530..2598,2599..3390)
                     /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      3561..4149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4050..4069
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4303..4320
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg aattgtgagc ggataacaat tcctctagaa ataattttgt
       61 ttaactttaa gaaggagata tatccatggg atccgaattc gcggccgccc taggctcgag
      121 ctaagcttga gtattctata gtgtcaccta aatcccagct tgatccggct gctaacaaag
      181 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      241 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataac
      301 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
      361 tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      421 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      481 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      541 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc acagcttgca
      601 tgcctgcagg tcggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactt
      661 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      721 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
      781 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
      841 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
      901 ggttgacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
      961 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     1021 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     1081 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     1141 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     1201 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     1261 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     1321 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     1381 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     1441 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     1501 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     1561 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     1621 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     1681 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     1741 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     1801 tgccataccg cgaaaggttt tgcaccattc gatggtgtcg ggtaccgagc tcgaatttac
     1861 gtgaaccatc acccaaatca agttttttgc ggtcgaggtg ccgtaaagct ctaaatcgga
     1921 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     1981 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2041 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcgcctgatg
     2101 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     2161 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     2221 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2281 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     2341 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     2401 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2461 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2521 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2581 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2641 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2701 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtcataca
     2761 ctattatccc gtattgacgc cgggcaagag caactcggtc gccgggcgcg gtattctcag
     2821 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2881 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2941 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3001 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3061 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3121 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3181 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3241 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3301 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3361 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3421 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3481 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3541 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3601 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3661 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3721 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3781 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3841 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3901 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3961 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4021 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4081 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4141 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4201 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4261 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4321 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4381 tgcagctgat
//
