LOCUS       Exported                6433 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Plasmid encoding the bacterial expression of a SNAP-fusion of p24 
            HIV Capsid protein.
ACCESSION   .
VERSION     .
KEYWORDS    pET51b(+)_SNAP-p24
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6433)
  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 6433)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6433
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(7..26)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     167..189
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(183..374)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(946..2028)
                     /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     1990..2009
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2029..2106)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2312..2331
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2419..2438
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2419..2437
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2438..2462
                     /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             2477..2499
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2515..2538
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2545..2559
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             3844..3873
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(3896..3914)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3910..3957
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3994..4449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4081..4100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4291..4312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4564..5424)
                     /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     5187..5206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5425..5517)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5598..6186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6087..6106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6340..6357
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6372..81
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
       61 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
      121 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
      181 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
      241 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
      301 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
      361 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
      421 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
      481 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
      541 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
      601 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
      661 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
      721 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
      781 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gctagtcatg
      841 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
      901 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
      961 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1021 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     1081 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     1141 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     1201 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     1261 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     1321 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     1381 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     1441 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     1501 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     1561 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     1621 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     1681 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     1741 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     1801 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     1861 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     1921 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     1981 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     2041 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     2101 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     2161 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     2221 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     2281 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     2341 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg
     2401 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
     2461 cccctctaga aataattttg tttaacttta agaaggagat ataccatggc aagctggagc
     2521 cacccgcagt tcgaaaaggg tgcagatgac gacgacaagg taccgcatat ggataaagac
     2581 tgcgaaatga aacgcactac cctggacagc ccgctgggta aactggaact gagcggttgc
     2641 gagcagggcc tgcacgaaat cattttcctg ggcaaaggta cgagcgctgc ggatgcggtc
     2701 gaagtaccgg ctccggctgc ggttctgggt ggcccggagc cgctgatgca ggctaccgca
     2761 tggctgaacg catattttca ccagccagag gcgattgaag agttccctgt accggccctg
     2821 catcacccgg tatttcagca ggaatccttt acccgtcagg tactgtggaa actgctgaaa
     2881 gtagtgaaat tcggcgaggt gatctcctat tcccatctgg cggcgctggc tggcaaccca
     2941 gcggcaactg cggcggtgaa aaccgctctg tctggtaatc cggtgccaat tctgatcccg
     3001 tgccaccgtg tggttcaggg cgacctggac gtaggtggct acgaaggcgg cctggctgtg
     3061 aaagaatggc tgctggcgca cgaaggtcac cgtctgggta aaccaggcct gggcagcggc
     3121 cctatagtgc agaacctcca ggggcaaatg gtacatcagg ccatatcacc tagaacttta
     3181 aatgcatggg taaaagtagt agaagagaag gctttcagcc cagaagtaat acccatgttt
     3241 tcagcattat cagaaggagc caccccacaa gatttaaata ccatgctaaa cacagtgggg
     3301 ggacatcaag cagccatgca aatgttaaaa gagaccatca atgaggaagc tgcagaatgg
     3361 gatagattgc atccagtgca tgcagggcct attgcaccag gccagatgag agaaccaagg
     3421 ggaagtgaca tagcaggaac tactagtacc cttcaggaac aaataggatg gatgacacat
     3481 aatccaccta tcccagtagg agaaatctat aaaagatgga taatcctggg attaaataaa
     3541 atagtaagaa tgtatagccc taccagcatt ctggacataa gacaaggacc aaaggaaccc
     3601 tttagagact atgtagaccg attctataaa actctaagag ccgagcaagc ttcacaagag
     3661 gtaaaaaatt ggatgacaga aaccttgttg gtccaaaatg cgaacccaga ttgtaagact
     3721 attttaaaag cattgggacc aggagcgaca ctagaagaaa tgatgacagc atgtcaggga
     3781 gtggggggac ccggccataa agcaagagtt ttgggagctc cgggcttctc ctcaatttcc
     3841 gctcatcacc accatcatca ccatcaccac cactaattaa cctaggctgc tgccaccgct
     3901 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     3961 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     4021 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4081 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4141 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4201 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4261 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4321 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4381 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     4441 ttacaatttc tggcggcacg atggcatgag attatcaaaa aggatcttca cctagatcct
     4501 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4561 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4621 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4681 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4741 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4801 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4861 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4921 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4981 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5041 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5101 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5161 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5221 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5281 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5341 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5401 gacacggaaa tgttgaatac tcatactctt cctttttcaa tcatgattga agcatttatc
     5461 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5521 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5581 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5641 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5701 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     5761 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5821 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5881 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5941 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6001 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6061 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6121 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6181 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6241 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     6301 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     6361 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     6421 atatatggtg cac
//
