LOCUS       Exported                5786 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Fluorophore-friendly streptavidin that can improve fluorescent 
            brightness and has unimpaired biotin binding after dye labeling.
ACCESSION   .
VERSION     .
KEYWORDS    pET21-Flavidin
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"
     primer_bind     114..133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     367..384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    399..538
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(464..483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     624..646
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(640..831)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1640..2722)
                     /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
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLVGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2684..2703
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2723..2800)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3006..3025
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3109..3128
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3109..3127
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3128..3152
                     /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             3167..3189
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3626..3643
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3696..3714)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3710..3757
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3794..4249
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3881..3900)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4091..4112
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4275..4379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4380..5240
                     /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(4598..4617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5411..213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
       61 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
      421 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
      481 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
      541 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
      601 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
      661 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
      721 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
      781 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
      841 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
      901 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
      961 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     1021 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     1081 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     1141 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     1201 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     1261 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     1321 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     1381 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     1441 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     1501 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     1561 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     1621 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1681 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     1741 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     1801 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     1861 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     1921 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     1981 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     2041 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     2101 tgcgagtgag atatttatgc cagccaacca gacgcagacg cgccgagaca gaacttaatg
     2161 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     2221 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     2281 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     2341 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     2401 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     2461 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     2521 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     2581 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     2641 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     2701 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     2761 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     2821 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     2881 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     2941 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     3001 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     3061 gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca
     3121 ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta
     3181 agaaggagat atacatatgg ctgaagctgg tatcaccggc acctggtaca accagctggg
     3241 atccaccttc atcgttaccg ctggtgctga cggtgctctg accggtacct acgaatccgc
     3301 tgttggtaac gctgaatcta gatacgttct gaccggtcgt tacgactccg ctccggctac
     3361 cgacggttcc ggaaccgctc tgggttggac cgttgcttgg cgcaacaact accgtaacgc
     3421 tcactccgct accacctggt ctggccagta cgttggtggt gctgaagctc gtatcaacac
     3481 ccagtggttg ttgacctccg gcaccaccga agctaacgcg tggcgatcca ccctggttgg
     3541 tcacgacacc ttcacccgtg ttcgtccgtc cgctgcttcc gaagaggaag aggaagagta
     3601 ataaaagctt gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa
     3661 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     3721 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     3781 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3841 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3901 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3961 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4021 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4081 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4141 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4201 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     4261 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4321 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4381 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4441 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4501 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4561 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4621 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4681 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4741 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4801 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4861 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4921 ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4981 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5041 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5101 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5161 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5221 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5281 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5341 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5401 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5461 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5521 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5581 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5641 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5701 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5761 agcgaacgac ctacaccgaa ctgaga
//
