LOCUS       sequence-254948-        6430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encoding the bacterial expression of a SNAP-fusion of p24 
            HIV Capsid protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6430)
  AUTHORS   Farrants H, Tarnawski M, Muller TG, Otsuka S, Hiblot J, Koch B, 
            Kueblbeck M, Krausslich HG, Ellenberg J, Johnsson K
  TITLE     Chemogenetic Control of Nanobodies.
  JOURNAL   Nat Methods. 2020 Feb 17. pii: 10.1038/s41592-020-0746-7. doi: 
            10.1038/s41592-020-0746-7.
  PUBMED    32066961
REFERENCE   2  (bases 1 to 6430)
  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"; journalName: "Nat 
            Methods. 2020 Feb 17. pii: 10.1038/s41592-020-0746-7. doi: 
            10.1038/s41592-020-0746-7."
COMMENT     Sequence Label: pET51b(+)_SNAP-p24
FEATURES             Location/Qualifiers
     source          1..6430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        23..41
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    42..66
                     /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             81..103
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             119..142
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             149..163
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             1448..1477
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1500..1518)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1514..1561
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1598..2053
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1685..1704)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1895..1916
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(2168..2959,2960..3028))
                     /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     2791..2810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3029..3121)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3202..3790
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3691..3710
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3944..3961
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3976..4115
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4041..4060)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4201..4223
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4217..4408)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4980..6062)
                     /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     6024..6043
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6063..6140)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6346..6365
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac
       61 aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacca tggcaagctg
      121 gagccacccg cagttcgaaa agggtgcaga tgacgacgac aaggtaccgc atatggataa
      181 agactgcgaa atgaaacgca ctaccctgga cagcccgctg ggtaaactgg aactgagcgg
      241 ttgcgagcag ggcctgcacg aaatcatttt cctgggcaaa ggtacgagcg ctgcggatgc
      301 ggtcgaagta ccggctccgg ctgcggttct gggtggcccg gagccgctga tgcaggctac
      361 cgcatggctg aacgcatatt ttcaccagcc agaggcgatt gaagagttcc ctgtaccggc
      421 cctgcatcac ccggtatttc agcaggaatc ctttacccgt caggtactgt ggaaactgct
      481 gaaagtagtg aaattcggcg aggtgatctc ctattcccat ctggcggcgc tggctggcaa
      541 cccagcggca actgcggcgg tgaaaaccgc tctgtctggt aatccggtgc caattctgat
      601 cccgtgccac cgtgtggttc agggcgacct ggacgtaggt ggctacgaag gcggcctggc
      661 tgtgaaagaa tggctgctgg cgcacgaagg tcaccgtctg ggtaaaccag gcctgggcag
      721 cggccctata gtgcagaacc tccaggggca aatggtacat caggccatat cacctagaac
      781 tttaaatgca tgggtaaaag tagtagaaga gaaggctttc agcccagaag taatacccat
      841 gttttcagca ttatcagaag gagccacccc acaagattta aataccatgc taaacacagt
      901 ggggggacat caagcagcca tgcaaatgtt aaaagagacc atcaatgagg aagctgcaga
      961 atgggataga ttgcatccag tgcatgcagg gcctattgca ccaggccaga tgagagaacc
     1021 aaggggaagt gacatagcag gaactactag tacccttcag gaacaaatag gatggatgac
     1081 acataatcca cctatcccag taggagaaat ctataaaaga tggataatcc tgggattaaa
     1141 taaaatagta agaatgtata gccctaccag cattctggac ataagacaag gaccaaagga
     1201 accctttaga gactatgtag accgattcta taaaactcta agagccgagc aagcttcaca
     1261 agaggtaaaa aattggatga cagaaacctt gttggtccaa aatgcgaacc cagattgtaa
     1321 gactatttta aaagcattgg gaccaggagc gacactagaa gaaatgatga cagcatgtca
     1381 gggagtgggg ggacccggcc ataaagcaag agttttggga gctccgggct tctcctcaat
     1441 ttccgctcat caccaccatc atcaccatca ccaccactaa ttaacctagg ctgctgccac
     1501 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1561 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     1621 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1681 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1741 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1801 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     1861 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     1921 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     1981 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2041 acgtttacaa tttctggcgg cacgatggca tgagattatc aaaaaggatc ttcacctaga
     2101 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2221 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2281 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2341 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2401 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2461 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2521 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2581 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2641 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2701 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2761 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2821 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2881 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2941 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3001 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatcatga ttgaagcatt
     3061 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3121 ataggtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3181 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3241 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3301 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3361 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3421 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3481 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3541 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3601 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3661 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3721 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3781 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3841 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3901 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3961 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4021 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     4081 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     4141 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4201 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     4261 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4321 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4381 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     4441 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     4501 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     4561 accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc
     4621 cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg
     4681 acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc
     4741 aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat
     4801 tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga
     4861 tcatgctagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4921 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4981 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5041 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     5101 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     5161 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     5221 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     5281 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5341 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5401 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5461 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5521 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5581 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5641 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5701 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5761 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5821 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5881 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5941 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     6001 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     6061 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     6121 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     6181 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     6241 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     6301 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     6361 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     6421 gaggatcgag
//
