LOCUS       Exported                7199 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Produces holo-PhyB* in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    pMH610
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7199)
  AUTHORS   Horner M, Gerhardt K, Salavei P, Hoess P, Harrer D, Kaiser J, Tabor 
            JJ, Weber W
  TITLE     Production of Phytochromes by High-Cell-Density E. coli 
            Fermentation.
  JOURNAL   ACS Synth Biol. 2019 Sep 26. doi: 10.1021/acssynbio.9b00267.
  PUBMED    31526004
REFERENCE   2  (bases 1 to 7199)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7199
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /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             2072..2089
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        2170..2188
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2189..2213
                     /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             2994..3005
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3784..3828
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3866..3884)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3880..3927
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4098..4889)
                     /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(4890..4981)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5029..5767)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(5977..7059)
                     /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     7021..7040
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7060..7137)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        7183..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggttagcg gtgttggtgg tagcggtggt ggtcgtggtg gcggtcgcgg
      121 aggtgaagaa gaaccgagca gcagccatac cccgaataat cgccgtggtg gtgaacaggc
      181 acagagcagc ggcaccaaaa gcctgcgtcc gcgtagcaat accgaaagca tgagcaaagc
      241 aattcagcag tataccgttg atgcacgtct gcatgccgtt ttcgaacaga gcggtgaaag
      301 cggtaaaagc tttgattata gccagagcct gaaaaccacc acctatggta gcagcgtgcc
      361 ggaacagcag attaccgcat atctgagccg tattcagcgt ggtggctaca ttcagccgtt
      421 tggctgcatg atcgcagttg atgaaagcag ctttcgcatt attggctaca gcgaaaatgc
      481 acgtgaaatg ctgggcatta tgccgcagag cgttccgacc ctggaaaaac cggaaattct
      541 ggcaatgggc accgatgttc gtagcctgtt taccagcagc agctccattc tgctggaacg
      601 tgcctttgtt gcccgtgaaa ttaccctgct gaatccggtt tggattcata gcaaaaacac
      661 cggcaaaccg ttttatgcaa ttctgcatcg tattgatgtt ggcgtggtta ttgatctgga
      721 accggcacgt accgaagatc cggcactgag cattgccggt gcagttcaga gccagaaact
      781 ggcagttcgt gcaattagcc agctgcaggc actgcctggt ggtgatatca aactgctgtg
      841 tgataccgtt gttgaaagcg ttcgtgatct gaccggctac gaccgtgtta tggtgtataa
      901 attccacgaa gatgaacatg gtgaagttgt tgcagaaagc aaacgtgatg acctggaacc
      961 gtatattggt ctgcattatc cagcaaccga tattccgcag gcaagccgtt tcctgttcaa
     1021 acagaatcgt gtgcgcatga ttgttgattg taatgcaaca ccggttctgg ttgttcagga
     1081 tgatcgtctg acccagagca tgtgtctggt tggtagcacc ctgcgtgcac cgcatggttg
     1141 tcatagccag tatatggcaa atatgggtag catcgcaagc ctggccatgg cggtgatcat
     1201 caatggtaat gaagatgatg gtagcaatgt tgcaagcggt cgtagcagca tgcgtctgtg
     1261 gggtctggtt gtgtgtcatc ataccagcag tcgctgcatt ccgtttccgc tgcgttatgc
     1321 atgtgaattt ctgatgcagg catttggact gcagctgaat atggaactgc aactggcact
     1381 gcagatgagc gaaaaacgtg ttctgcgtac ccagaccctg ctgtgcgata tgctgctgcg
     1441 tgatagtccg gcaggcattg ttacccagag cccgagcatt atggatctgg tgaaatgcga
     1501 tggtgcagcc tttctgtatc acggtaaata ctatccgctg ggtgttgcac cgagcgaagt
     1561 tcagattaaa gatgttgttg agtggctgct ggcaaatcat gcagatagca ccggtctgag
     1621 caccgatagc ctgggtgatg caggttatcc gggtgcagca gcactgggag atgcagtttg
     1681 tggtatggca gttgcataca ttaccaaacg cgattttctg ttttggtttc gtagccatac
     1741 cgccaaagaa atcaaatggg gtggtgcaaa acatcacccg gaagataaag atgacggtca
     1801 gcgtatgcat ccgcgtagta gctttcaggc atttctggaa gtggtgaaaa gccgtagcca
     1861 gccgtgggaa accgcagaaa tggatgcaat tcatagcctg caactgattc tgcgcgatag
     1921 cttcaaagaa agcgaagcag caatgaatag caaagttgtt gatggtgttg ttcagccgtg
     1981 tcgtgatatg gcaggcgaac agggtattga tgaactgggt gcaggttctg gtagcggtcg
     2041 tggcgatagc ccaggtcgtg gtgactctcc gcatcatcac catcaccatg atcgctgtgg
     2101 ttaagcggcc gcataatgct taagtcgaac agaaagtaat cgtattgtac acggccgcat
     2161 aatcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tccccatctt
     2221 agtatattag ttaagtataa gaaggagata tacatatgag tgtcaactta gcttcccagt
     2281 tgcgggaagg gacgaaaaaa tcccactcca tggcggagaa cgtcggcttt gtcaaatgct
     2341 tcctcaaggg cgttgtcgag aaaaattcct accgtaagct ggttggcaat ctctactttg
     2401 tctacagtgc catggaagag gaaatggcaa aatttaagga ccatcccatc ctcagccaca
     2461 tttacttccc cgaactcaac cgcaaacaaa gcctagagca agacctgcaa ttctattacg
     2521 gctccaactg gcggcaagaa gtgaaaattt ctgccgctgg ccaagcctat gtggaccgag
     2581 tccggcaagt ggccgctacg gcccctgaat tgttggtggc ccattcctac acccgttacc
     2641 tgggggatct ttccggcggt caaattctca agaaaattgc ccaaaatgcc atgaatctcc
     2701 acgatggtgg cacagctttc tatgaatttg ccgacattga tgacgaaaag gcttttaaaa
     2761 atacctaccg tcaagctatg aatgatctgc ccattgacca agccaccgcc gaacggattg
     2821 tggatgaagc caatgacgcc tttgccatga acatgaaaat gttcaacgaa cttgaaggca
     2881 acctgatcaa ggcgatcggc attatggtgt tcaacagcct cacccgtcgc cgcagtcaag
     2941 gcagcaccga agttggcctc gccacctccg aaggctagaa gcttggatcc attaaagagg
     3001 agaaacccgg gcatggccgt cactgattta agtttgacca attcttccct gatgcctacg
     3061 ttgaacccga tgattcaaca gttggccctg gcgatcgccg ctagttggca aagtttaccc
     3121 ctcaagccct atcaattgcc ggaggatttg ggctacgtag aaggccgcct ggaaggggaa
     3181 aagttagtga ttgaaaatcg gtgctaccaa acgccccagt ttcgcaaaat gcatttggag
     3241 ttggccaagg tgggcaaagg gttggatatt ctccactgtg taatgtttcc tgagccttta
     3301 tacggtctac ctttgtttgg ctgtgacatt gtggccggcc ccggtggagt aagtgcggct
     3361 attgcggatc tatcccccac ccaaagcgat cgccaattgc ccgcagcgta ccaaaaatca
     3421 ttggcagagc taggccagcc agaatttgag caacaacggg aattgccccc ctggggagaa
     3481 atattttctg aatattgttt attcatccgt cccagcaatg tcactgaaga agaaagattt
     3541 gtacaaaggg tagtggactt tttgcaaatt cattgtcacc aatccatcgt tgccgaaccc
     3601 ttgtctgaag ctcaaacttt ggagcaccgt caggggcaaa ttcattactg ccaacaacaa
     3661 cagaaaaatg ataaaacccg tcgggtactg gaaaaagctt ttggggaagc ttgggcggaa
     3721 cggtatatga gccaagtctt atttgatgtt atccaataag acgtcggtac cctcgagtct
     3781 ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca
     3841 gcttaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     3901 tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc
     3961 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
     4021 cgaccctgcc ctgaaccgac gaccgggtca tcgtggccgg atcttgcggc ccctcggctt
     4081 gaacgaattg ttagacatta tttgccgact accttggtga tctcgccttt cacgtagtgg
     4141 acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa
     4201 gcctgtctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
     4261 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
     4321 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
     4381 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
     4441 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
     4501 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
     4561 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
     4621 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
     4681 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
     4741 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
     4801 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
     4861 gatacttcgg cgatcaccgc ttccctcata ctcttccttt ttcaatatta ttgaagcatt
     4921 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4981 atagctagct cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca
     5041 catacaaagt tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca
     5101 gcagggccgc gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa
     5161 acagacgctt ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta
     5221 cgggcgaaac ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg
     5281 cgctctcctg ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg
     5341 gaagtgtggc gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg
     5401 ctccaagctg ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg
     5461 taactgttca cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat
     5521 tggtaactgg gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt
     5581 gcgccaaagt ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag
     5641 ttaccacggt taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt
     5701 ggttttttcg tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc
     5761 tttgatcttt tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc
     5821 acctgaagtc agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc
     5881 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     5941 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6001 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6061 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     6121 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     6181 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     6241 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     6301 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     6361 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     6421 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     6481 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     6541 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     6601 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     6661 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     6721 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     6781 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     6841 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6901 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6961 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     7021 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     7081 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     7141 gatctcgacg ctctccctta tgcgactcct gcattaggaa attaatacga ctcactata
//
