LOCUS       sequence-212061-        4468 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 4468)
  TITLE     Hyvonen Lab pOP plasmids
REFERENCE   2  (bases 1 to 4468)
  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..4468
                     /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             122..145
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             152..196
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     promoter        complement(213..231)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(214..231)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(289..307)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      303..350
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(586..607)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(757..1839)
                     /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     1801..1820
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1840..1917)
                     /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     2063..2082
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(2238..2257)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2357..2379
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2417..2435)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2503..2607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2608..2676,2677..3468)
                     /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      3639..4227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4128..4147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4381..4398
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg aattgtgagc ggataacaat tcctctagaa ataattttgt
       61 ttaactttaa gaaggagata tatccatggg atccgaattc gcggccgccc taggctcgag
      121 ccatcaccat caccatcacc atcacactag tggactgaat gacattttcg aagcacagaa
      181 gatcgaatgg catgaagcct aagcttgagt attctatagt gtcacctaaa tcccagcttg
      241 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      301 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      361 gaactatatc cggataacct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      421 acagttgcgc agcctgaatg gcgaatggaa attgtaagcg ttaatatttt gttaaaattc
      481 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
      541 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
      601 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
      661 gatggcccac agcttgcatg cctgcaggtc ggaagcataa agtgtaaagc ctggggtgcc
      721 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      781 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
      841 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
      901 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
      961 aaaatcctgt ttgatggtgg ttgacggcgg gatataacat gagctgtctt cggtatcgtc
     1021 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     1081 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     1141 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     1201 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     1261 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     1321 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     1381 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     1441 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     1501 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     1561 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     1621 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     1681 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     1741 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     1801 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     1861 ctcttccggg cgctatcatg ccataccgcg aaaggttttg caccattcga tggtgtcggg
     1921 taccgagctc gaatttacgt gaaccatcac ccaaatcaag ttttttgcgg tcgaggtgcc
     1981 gtaaagctct aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2041 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2101 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2161 agggcgcgtc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     2221 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     2281 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2341 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2401 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     2461 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2521 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2581 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2641 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2701 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2761 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2821 gttctgctat gtcatacact attatcccgt attgacgccg ggcaagagca actcggtcgc
     2881 cgggcgcggt attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2941 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3001 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3061 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3121 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3181 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3241 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3301 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3361 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3421 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3481 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3541 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3601 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3661 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3721 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3781 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3841 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3901 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3961 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4021 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4081 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4141 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4201 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4261 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4321 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4381 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4441 cgttggccga ttcattaatg cagctgat
//
