LOCUS       sequence-258490-        6831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces holo-Cph1* in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6831)
  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 6831)
  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: "ACS Synth 
            Biol. 2019 Sep 26. doi: 10.1021/acssynbio.9b00267."
COMMENT     Sequence Label: pMH48
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             1665..1682
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1754..1772
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1773..1797
                     /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             2578..2589
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3368..3412
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3450..3468)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3464..3511
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3682..4473)
                     /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(4474..4565)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(5626..6708)
                     /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     6670..6689
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6709..6786)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taatcagaag gaatacatgg ccaccaccgt acaactcagc gaccaatccc
      121 tccgtcagct agaaaccctc gccatccaca ccgcccacct gattcagccc cacggtttag
      181 tggtggtcct gcaggaacca gacctcacca tcagccaaat tagcgccaac tgcaccggca
      241 ttttagggcg atcgccagag gatttgttgg gcagaaccct aggggaagtg tttgatagct
      301 ttcagattga tcccatccag agtcgcctaa cggccggaca aatcagcagc ctcaacccca
      361 gtaaactttg ggcgcgggtc atgggggacg actttgtcat ttttgacggg gtttttcatc
      421 gcaacagtga cggtttattg gtatgtgaac tagagccagc ctacacttcc gataatctgc
      481 ccttcctcgg tttttatcac atggccaacg ctgccctgaa tcggttgcgc caacaagcta
      541 atctacggga tttctacgat gttattgtcg aagaagtccg ccgtatgact ggctttgacc
      601 gggtgatgct ataccgcttt gatgaaaata accacggtga tgtcattgcc gaagataaac
      661 gggatgatat ggaaccctat ttgggcctgc actatcccga atcggatatt ccccaacccg
      721 cccgtcggct atttatccac aaccccattc gagtaattcc cgatgtttat ggtgtggcgg
      781 tgcccctgac cccagcggtt aaccccagca ccaaccgagc ggtggattta acagaatcca
      841 ttctgcgcag tgcgtaccat tgccacttga cctttctgaa aaatatgggg gtaggagcgt
      901 ctttaaccat ttccctaatt aaggacggcc atctctgggg gctcattgcc tgccaccatc
      961 aaacccccaa agtaattccc tttgaactgc gtaaagcctg cgaatttttt ggtcgggtgg
     1021 tgtttagcaa catttccgcc caggaagata cggaaacctt cgattaccgg gtgcagttgg
     1081 cggagcatga agcggtttta ttggacaaaa tgaccacggc ggcggatttt gtcgaaggat
     1141 taactaatca tcccgatcgc ctgttgggat taacgggctc ccagggggcg gccatttgct
     1201 ttggggaaaa attgatttta gtaggggaaa ccccggacga gaaagcagtg caatatttac
     1261 tgcaatggtt ggagaatcgg gaagtgcaag acgttttctt cacctcttcc ctctcacaaa
     1321 tttatcctga tgcagtgaat tttaaatccg tggccagtgg cttattggcc attcccattg
     1381 cccgtcacaa ctttttgctc tggtttcgcc ctgaagtgtt gcaaacggtt aattggggcg
     1441 gtgacccaaa tcatgcttac gaagctaccc aggaagacgg taaaatcgag ctccatcccc
     1501 gccaatcctt tgacctctgg aaagaaattg tccgactcca atctttgccc tggcaatcgg
     1561 tggaaatcca aagtgccctg gccctgaaaa aggcgatcgt caacctcatt ttgcgccagg
     1621 cagaagaagg ccgtggcgat agcccaggtc gtggtgactc tccgcatcat caccatcacc
     1681 attgctgagc ggccgcataa tgcttaagtc gaacagaaag taatcgtatt gtacacggcc
     1741 gcataatcga aattaatacg actcactata ggggaattgt gagcggataa caattcccca
     1801 tcttagtata ttagttaagt ataagaagga gatatacata tgagtgtcaa cttagcttcc
     1861 cagttgcggg aagggacgaa aaaatcccac tccatggcgg agaacgtcgg ctttgtcaaa
     1921 tgcttcctca agggcgttgt cgagaaaaat tcctaccgta agctggttgg caatctctac
     1981 tttgtctaca gtgccatgga agaggaaatg gcaaaattta aggaccatcc catcctcagc
     2041 cacatttact tccccgaact caaccgcaaa caaagcctag agcaagacct gcaattctat
     2101 tacggctcca actggcggca agaagtgaaa atttctgccg ctggccaagc ctatgtggac
     2161 cgagtccggc aagtggccgc tacggcccct gaattgttgg tggcccattc ctacacccgt
     2221 tacctggggg atctttccgg cggtcaaatt ctcaagaaaa ttgcccaaaa tgccatgaat
     2281 ctccacgatg gtggcacagc tttctatgaa tttgccgaca ttgatgacga aaaggctttt
     2341 aaaaatacct accgtcaagc tatgaatgat ctgcccattg accaagccac cgccgaacgg
     2401 attgtggatg aagccaatga cgcctttgcc atgaacatga aaatgttcaa cgaacttgaa
     2461 ggcaacctga tcaaggcgat cggcattatg gtgttcaaca gcctcacccg tcgccgcagt
     2521 caaggcagca ccgaagttgg cctcgccacc tccgaaggct agaagcttgg atccattaaa
     2581 gaggagaaac ccgggcatgg ccgtcactga tttaagtttg accaattctt ccctgatgcc
     2641 tacgttgaac ccgatgattc aacagttggc cctggcgatc gccgctagtt ggcaaagttt
     2701 acccctcaag ccctatcaat tgccggagga tttgggctac gtagaaggcc gcctggaagg
     2761 ggaaaagtta gtgattgaaa atcggtgcta ccaaacgccc cagtttcgca aaatgcattt
     2821 ggagttggcc aaggtgggca aagggttgga tattctccac tgtgtaatgt ttcctgagcc
     2881 tttatacggt ctacctttgt ttggctgtga cattgtggcc ggccccggtg gagtaagtgc
     2941 ggctattgcg gatctatccc ccacccaaag cgatcgccaa ttgcccgcag cgtaccaaaa
     3001 atcattggca gagctaggcc agccagaatt tgagcaacaa cgggaattgc ccccctgggg
     3061 agaaatattt tctgaatatt gtttattcat ccgtcccagc aatgtcactg aagaagaaag
     3121 atttgtacaa agggtagtgg actttttgca aattcattgt caccaatcca tcgttgccga
     3181 acccttgtct gaagctcaaa ctttggagca ccgtcagggg caaattcatt actgccaaca
     3241 acaacagaaa aatgataaaa cccgtcgggt actggaaaaa gcttttgggg aagcttgggc
     3301 ggaacggtat atgagccaag tcttatttga tgttatccaa taagacgtcg gtaccctcga
     3361 gtctggtaaa gaaaccgctg ctgcgaaatt tgaacgccag cacatggact cgtctactag
     3421 cgcagcttaa ttaacctagg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     3481 ggcctctaaa cgggtcttga ggggtttttt gctgaaacct caggcatttg agaagcacac
     3541 ggtcacactg cttccggtag tcaataaacc ggtaaaccag caatagacat aagcggctat
     3601 ttaacgaccc tgccctgaac cgacgaccgg gtcatcgtgg ccggatcttg cggcccctcg
     3661 gcttgaacga attgttagac attatttgcc gactaccttg gtgatctcgc ctttcacgta
     3721 gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag
     3781 ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc
     3841 ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg
     3901 ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag
     3961 cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc
     4021 ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc
     4081 cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca
     4141 ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac
     4201 aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc
     4261 caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac
     4321 cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg
     4381 tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg
     4441 agtcgatact tcggcgatca ccgcttccct catactcttc ctttttcaat attattgaag
     4501 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4561 acaaatagct agctcactcg gtcgctacgc tccgggcgtg agactgcggc gggcgctgcg
     4621 gacacataca aagttaccca cagattccgt ggataagcag gggactaaca tgtgaggcaa
     4681 aacagcaggg ccgcgccggt ggcgtttttc cataggctcc gccctcctgc cagagttcac
     4741 ataaacagac gcttttccgg tgcatctgtg ggagccgtga ggctcaacca tgaatctgac
     4801 agtacgggcg aaacccgaca ggacttaaag atccccaccg tttccggcgg gtcgctccct
     4861 cttgcgctct cctgttccga ccctgccgtt taccggatac ctgttccgcc tttctccctt
     4921 acgggaagtg tggcgctttc tcatagctca cacactggta tctcggctcg gtgtaggtcg
     4981 ttcgctccaa gctgggctgt aagcaagaac tccccgttca gcccgactgc tgcgccttat
     5041 ccggtaactg ttcacttgag tccaacccgg aaaagcacgg taaaacgcca ctggcagcag
     5101 ccattggtaa ctgggagttc gcagaggatt tgtttagcta aacacgcggt tgctcttgaa
     5161 gtgtgcgcca aagtccggct acactggaag gacagatttg gttgctgtgc tctgcgaaag
     5221 ccagttacca cggttaagca gttccccaac tgacttaacc ttcgatcaaa ccacctcccc
     5281 aggtggtttt ttcgaaggtt aagtcagttg gggaactgct taaccttcga tcaaaccacc
     5341 tccccaggtg gttttttcgt ttacagggca aaagattacg cgcagaaaaa aaggatctca
     5401 agaagatcct ttgatctttt ctactgaacc gctctagatt tcagtgcaat ttatctcttc
     5461 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gttagtcatg
     5521 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     5581 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     5641 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5701 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     5761 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5821 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     5881 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     5941 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     6001 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     6061 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     6121 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     6181 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     6241 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     6301 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     6361 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     6421 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     6481 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     6541 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     6601 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     6661 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     6721 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     6781 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaaa t
//
