LOCUS       Exported                5786 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Streptavidin that has unimpaired biotin binding after dye labeling.
ACCESSION   .
VERSION     .
KEYWORDS    pET21-Streptavidin-K121R
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5786)
  AUTHORS   Jacobsen MT, Fairhead M, Fogelstrand P, Howarth M
  TITLE     Amine Landscaping to Maximize Protein-Dye Fluorescence and 
            Ultrastable Protein-Ligand Interaction.
  JOURNAL   Cell Chem Biol. 2017 Jul 17. pii: S2451-9456(17)30229-5. doi: 
            10.1016/j.chembiol.2017.06.015.
  PUBMED    28757182
REFERENCE   2  (bases 1 to 5786)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5786
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      116..704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     605..624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     858..875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    890..1029
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(955..974)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1115..1137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1131..1322)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2131..3213)
                     /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     3175..3194
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3214..3291)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3497..3516
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3600..3619
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3600..3618
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3619..3643
                     /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             3658..3680
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4117..4134
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4187..4205)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4201..4248
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4285..4740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4372..4391)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4582..4603
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4766..4870
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4871..5731
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5089..5108)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
       61 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      241 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
      301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
      841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
      901 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
      961 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     1021 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     1081 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     1141 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     1201 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     1261 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     1321 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     1381 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     1441 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     1501 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     1561 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     1621 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     1681 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     1741 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     1801 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     1861 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     1921 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     1981 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     2041 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     2101 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2161 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2221 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     2281 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     2341 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     2401 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     2461 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     2521 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     2581 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     2641 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     2701 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     2761 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     2821 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     2881 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     2941 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     3001 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     3061 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     3121 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     3181 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     3241 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     3301 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     3361 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     3421 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     3481 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     3541 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
     3601 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     3661 gtttaacttt aagaaggaga tatacatatg gctgaagctg gtatcaccgg cacctggtac
     3721 aaccagctgg gatccacctt catcgttacc gctggtgctg acggtgctct gaccggtacc
     3781 tacgaatccg ctgttggtaa cgctgaatct agatacgttc tgaccggtcg ttacgactcc
     3841 gctccggcta ccgacggttc cggaaccgct ctgggttgga ccgttgcttg gaaaaacaac
     3901 taccgtaacg ctcactccgc taccacctgg tctggccagt acgttggtgg tgctgaagct
     3961 cgtatcaaca cccagtggtt gttgacctcc ggcaccaccg aagctaacgc gtggcgatcc
     4021 accctggttg gtcacgacac cttcaccaaa gttaaaccgt ccgctgcttc cgaagaggaa
     4081 gaggaagagt aataaaagct tgcggccgca ctcgagcacc accaccacca ccactgagat
     4141 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     4201 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     4261 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4321 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4381 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4441 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4501 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4561 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4621 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     4681 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     4741 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4801 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4861 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4921 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4981 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5041 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5101 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5161 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5221 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5281 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5341 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5401 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5461 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5521 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5581 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5641 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5701 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5761 ttagattgat ttaaaacttc attttt
//
