LOCUS       sequence-258491-        7261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces holo-PhyB-AviTag in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7261)
  AUTHORS   Horner M, Yousefi OS, Schamel WWA, Weber W
  TITLE     Production, Purification and Characterization of Recombinant 
            Biotinylated Phytochrome B for Extracellular Optogenetics.
  JOURNAL   Bio Protoc. 2020 Mar 5;10(5):e3541. doi: 10.21769/BioProtoc.3541. 
            eCollection 2020 Mar 5.
  PUBMED    33659515
REFERENCE   2  (bases 1 to 7261)
  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: "Bio 
            Protoc."; date: "2020-03-5"; volume: "10(5):e3541. doi"; pages: " 
            10.21769/BioProtoc.3541. eCollection 2020 Mar 5"
COMMENT     Sequence Label: pMH1105
FEATURES             Location/Qualifiers
     source          1..7261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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)."
     CDS             2034..2078
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2079..2096
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2165..2183
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2184..2208
                     /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             2989..3000
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3779..3823
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3861..3879)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3875..3922
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4093..4884)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4885..4976)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(6037..7119)
                     /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     7081..7100
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7120..7197)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        7243..7261
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctgt agaaataatt ttgtttaact ttaataagga
       61 gatataccat ggttagcggt gttggtggta gcggtggtgg tcgtggtggc ggtcgcggag
      121 gtgaagaaga accgagcagc agccataccc cgaataatcg ccgtggtggt gaacaggcac
      181 agagcagcgg caccaaaagc ctgcgtccgc gtagcaatac cgaaagcatg agcaaagcaa
      241 ttcagcagta taccgttgat gcacgtctgc atgccgtttt cgaacagagc ggtgaaagcg
      301 gtaaaagctt tgattatagc cagagcctga aaaccaccac ctatggtagc agcgtgccgg
      361 aacagcagat taccgcatat ctgagccgta ttcagcgtgg tggctacatt cagccgtttg
      421 gctgcatgat cgcagttgat gaaagcagct ttcgcattat tggctacagc gaaaatgcac
      481 gtgaaatgct gggcattatg ccgcagagcg ttccgaccct ggaaaaaccg gaaattctgg
      541 caatgggcac cgatgttcgt agcctgttta ccagcagcag ctccattctg ctggaacgtg
      601 cctttgttgc ccgtgaaatt accctgctga atccggtttg gattcatagc aaaaacaccg
      661 gcaaaccgtt ttatgcaatt ctgcatcgta ttgatgttgg cgtggttatt gatctggaac
      721 cggcacgtac cgaagatccg gcactgagca ttgccggtgc agttcagagc cagaaactgg
      781 cagttcgtgc aattagccag ctgcaggcac tgcctggtgg tgatatcaaa ctgctgtgtg
      841 ataccgttgt tgaaagcgtt cgtgatctga ccggctacga ccgtgttatg gtgtataaat
      901 tccacgaaga tgaacatggt gaagttgttg cagaaagcaa acgtgatgac ctggaaccgt
      961 atattggtct gcattatcca gcaaccgata ttccgcaggc aagccgtttc ctgttcaaac
     1021 agaatcgtgt gcgcatgatt gttgattgta atgcaacacc ggttctggtt gttcaggatg
     1081 atcgtctgac ccagagcatg tgtctggttg gtagcaccct gcgtgcaccg catggttgtc
     1141 atagccagta tatggcaaat atgggtagca tcgcaagcct ggccatggcg gtgatcatca
     1201 atggtaatga agatgatggt agcaatgttg caagcggtcg tagcagcatg cgtctgtggg
     1261 gtctggttgt gtgtcatcat accagcagtc gctgcattcc gtttccgctg cgttatgcat
     1321 gtgaatttct gatgcaggca tttggactgc agctgaatat ggaactgcaa ctggcactgc
     1381 agatgagcga aaaacgtgtt ctgcgtaccc agaccctgct gtgcgatatg ctgctgcgtg
     1441 atagtccggc aggcattgtt acccagagcc cgagcattat ggatctggtg aaatgcgatg
     1501 gtgcagcctt tctgtatcac ggtaaatact atccgctggg tgttgcaccg agcgaagttc
     1561 agattaaaga tgttgttgag tggctgctgg caaatcatgc agatagcacc ggtctgagca
     1621 ccgatagcct gggtgatgca ggttatccgg gtgcagcagc actgggagat gcagtttgtg
     1681 gtatggcagt tgcatacatt accaaacgcg attttctgtt ttggtttcgt agccataccg
     1741 ccaaagaaat caaatggggt ggtgcaaaac atcacccgga agataaagat gacggtcagc
     1801 gtatgcatcc gcgtagtagc tttcaggcat ttctggaagt ggtgaaaagc cgtagccagc
     1861 cgtgggaaac cgcagaaatg gatgcaattc atagcctgca actgattctg cgcgatagct
     1921 tcaaagaaag cgaagcagca atgaatagca aagttgttga tggtgttgtt cagccgtgtc
     1981 gtgatatggc aggcgaacag ggtattgatg aactgggtgc aggttctggt agcggtctga
     2041 acgacatctt cgaagctcag aaaatcgaat ggcacgaaca tcatcaccat caccattaag
     2101 cggccgcata atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc cgcataatcg
     2161 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc atcttagtat
     2221 attagttaag tataagaagg agatatacat atgagtgtca acttagcttc ccagttgcgg
     2281 gaagggacga aaaaatccca ctccatggcg gagaacgtcg gctttgtcaa atgcttcctc
     2341 aagggcgttg tcgagaaaaa ttcctaccgt aagctggttg gcaatctcta ctttgtctac
     2401 agtgccatgg aagaggaaat ggcaaaattt aaggaccatc ccatcctcag ccacatttac
     2461 ttccccgaac tcaaccgcaa acaaagccta gagcaagacc tgcaattcta ttacggctcc
     2521 aactggcggc aagaagtgaa aatttctgcc gctggccaag cctatgtgga ccgagtccgg
     2581 caagtggccg ctacggcccc tgaattgttg gtggcccatt cctacacccg ttacctgggg
     2641 gatctttccg gcggtcaaat tctcaagaaa attgcccaaa atgccatgaa tctccacgat
     2701 ggtggcacag ctttctatga atttgccgac attgatgacg aaaaggcttt taaaaatacc
     2761 taccgtcaag ctatgaatga tctgcccatt gaccaagcca ccgccgaacg gattgtggat
     2821 gaagccaatg acgcctttgc catgaacatg aaaatgttca acgaacttga aggcaacctg
     2881 atcaaggcga tcggcattat ggtgttcaac agcctcaccc gtcgccgcag tcaaggcagc
     2941 accgaagttg gcctcgccac ctccgaaggc tagaagcttg gatccattaa agaggagaaa
     3001 cccgggcatg gccgtcactg atttaagttt gaccaattct tccctgatgc ctacgttgaa
     3061 cccgatgatt caacagttgg ccctggcgat cgccgctagt tggcaaagtt tacccctcaa
     3121 gccctatcaa ttgccggagg atttgggcta cgtagaaggc cgcctggaag gggaaaagtt
     3181 agtgattgaa aatcggtgct accaaacgcc ccagtttcgc aaaatgcatt tggagttggc
     3241 caaggtgggc aaagggttgg atattctcca ctgtgtaatg tttcctgagc ctttatacgg
     3301 tctacctttg tttggctgtg acattgtggc cggccccggt ggagtaagtg cggctattgc
     3361 ggatctatcc cccacccaaa gcgatcgcca attgcccgca gcgtaccaaa aatcattggc
     3421 agagctaggc cagccagaat ttgagcaaca acgggaattg cccccctggg gagaaatatt
     3481 ttctgaatat tgtttattca tccgtcccag caatgtcact gaagaagaaa gatttgtaca
     3541 aagggtagtg gactttttgc aaattcattg tcaccaatcc atcgttgccg aacccttgtc
     3601 tgaagctcaa actttggagc accgtcaggg gcaaattcat tactgccaac aacaacagaa
     3661 aaatgataaa acccgtcggg tactggaaaa agcttttggg gaagcttggg cggaacggta
     3721 tatgagccaa gtcttatttg atgttatcca ataagacgtc ggtaccctcg agtctggtaa
     3781 agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta
     3841 attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     3901 acgggtcttg aggggttttt tgctgaaacc tcaggcattt gagaagcaca cggtcacact
     3961 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     4021 ctgccctgaa ccgacgaccg ggtcatcgtg gccggatctt gcggcccctc ggcttgaacg
     4081 aattgttaga cattatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
     4141 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
     4201 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
     4261 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
     4321 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     4381 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     4441 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     4501 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     4561 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     4621 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     4681 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     4741 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     4801 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     4861 ttcggcgatc accgcttccc tcatactctt cctttttcaa tattattgaa gcatttatca
     4921 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagc
     4981 tagctcactc ggtcgctacg ctccgggcgt gagactgcgg cgggcgctgc ggacacatac
     5041 aaagttaccc acagattccg tggataagca ggggactaac atgtgaggca aaacagcagg
     5101 gccgcgccgg tggcgttttt ccataggctc cgccctcctg ccagagttca cataaacaga
     5161 cgcttttccg gtgcatctgt gggagccgtg aggctcaacc atgaatctga cagtacgggc
     5221 gaaacccgac aggacttaaa gatccccacc gtttccggcg ggtcgctccc tcttgcgctc
     5281 tcctgttccg accctgccgt ttaccggata cctgttccgc ctttctccct tacgggaagt
     5341 gtggcgcttt ctcatagctc acacactggt atctcggctc ggtgtaggtc gttcgctcca
     5401 agctgggctg taagcaagaa ctccccgttc agcccgactg ctgcgcctta tccggtaact
     5461 gttcacttga gtccaacccg gaaaagcacg gtaaaacgcc actggcagca gccattggta
     5521 actgggagtt cgcagaggat ttgtttagct aaacacgcgg ttgctcttga agtgtgcgcc
     5581 aaagtccggc tacactggaa ggacagattt ggttgctgtg ctctgcgaaa gccagttacc
     5641 acggttaagc agttccccaa ctgacttaac cttcgatcaa accacctccc caggtggttt
     5701 tttcgaaggt taagtcagtt ggggaactgc ttaaccttcg atcaaaccac ctccccaggt
     5761 ggttttttcg tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc
     5821 tttgatcttt tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc
     5881 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc
     5941 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     6001 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6061 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6121 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     6181 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     6241 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     6301 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     6361 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     6421 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     6481 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     6541 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     6601 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     6661 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     6721 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     6781 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     6841 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     6901 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6961 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     7021 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     7081 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     7141 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     7201 gatctcgacg ctctccctta tgcgactcct gcattaggaa attaatacga ctcactatag
     7261 g
//
