LOCUS       sequence-220206-        4463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of protein in E. coli as non-cleavable 
            C-terminal 8His-Avi tag fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4463)
  TITLE     Hyvonen Lab pOP plasmids
REFERENCE   2  (bases 1 to 4463)
  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: pOP2HA
FEATURES             Location/Qualifiers
     source          1..4463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..28
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9..27
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    27..51
                     /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             64..86
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             130..153
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             160..204
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     promoter        complement(221..239)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(222..239)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(297..315)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      311..358
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(594..615)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(765..1847)
                     /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     1809..1828
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1848..1925)
                     /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     2068..2087
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2241..2260)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2360..2382
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2420..2438)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2506..2610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2611..2679,2680..3471)
                     /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(2829..2848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3642..4230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4131..4150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4384..4401
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctgatta atacgactca ctatagggaa ttgtgagcgg ataacaattc ctctagaaat
       61 aattttgttt aactttaaga aggagatata tccatgggat ccgaattcgc ggccgcccta
      121 ggctcgagcc atcaccatca ccatcaccat cacactagtg gactgaatga cattttcgaa
      181 gcacagaaga tcgaatggca tgaagcctaa gcttgagtat tctatagtgt cacctaaatc
      241 ccagcttgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
      301 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      361 gaaaggagga actatatccg gataacctgg cgtaatagcg aagaggcccg caccgatcgc
      421 ccttcccaac agttgcgcag cctgaatggc gaatggaaat tgtaagcgtt aatattttgt
      481 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
      541 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
      601 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
      661 atcagggcga tggcccacag cttgcatgcc tgcaggtcgg aagcataaag tgtaaagcct
      721 ggggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
      781 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      841 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
      901 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
      961 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     1021 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     1081 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     1141 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     1201 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     1261 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     1321 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     1381 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     1441 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     1501 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     1561 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     1621 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     1681 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     1741 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     1801 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     1861 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca ccattcgatg
     1921 gtgtcgggta ccgagctcga attgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1981 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2041 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2101 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2161 gctacagggc gcgtcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2221 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     2281 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     2341 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     2401 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     2461 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2521 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2581 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2641 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2701 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2761 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2821 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2881 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2941 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3001 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3061 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3121 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3181 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3241 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3301 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3361 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3421 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3481 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3541 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3601 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3661 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3721 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3781 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3841 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3901 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3961 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4021 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4081 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4141 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4201 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4261 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4321 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4381 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4441 gcgcgttggc cgattcatta atg
//
