LOCUS       sequence-468087-        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For far-red light induced bacterial expression of gene of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7871)
  AUTHORS   Bohm C, Lehtinen K, Multamaki E, Vanhatalo R, Brander O, Meier SSM, 
            Rumfeldt J, Moglich A, Takala H
  TITLE     Traits of Bathy Phytochromes and Application to Bacterial 
            Optogenetics.
  JOURNAL   ACS Synth Biol. 2025 Jul 11. doi: 10.1021/acssynbio.5c00337.
  PUBMED    40645610
REFERENCE   2  (bases 1 to 7871)
  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. 2025 Jul 11. doi: 10.1021/acssynbio.5c00337."
COMMENT     Sequence Label: pFREDusk-KanR-MCS
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(205..237)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(241..258)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(268..285)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             304..326
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(655..674)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        880..957
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5170..5361
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5355..5377)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5463..5602
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5518..5537
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5617..5634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5788..6376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5868..5887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6487..6506)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6498..7313
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             6498..7313
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6571..6590)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(7405..7860)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7542..7563)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7754..7773
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgacccat ttgctgtcca ccagtcatgc tagccatatg
      241 gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt
      301 atatctcctt cttaaagtta aacaaaatta tttctagact ccgttgtgat gacgcattgg
      361 tacgcggtat cgggaggttc gaaaatttcg agcgatatct taaggggggt gccttacgta
      421 gaaccccgta ggtcatgccc gaggccggtc ctggatggcg cggcggatac gcttgagcag
      481 gttttcgtcg agaagcggct tcaaaaccac gtcttttacg ccggcctcgg cggcccgggt
      541 cgagatgttt tcgtccggat agccggtgat caggatcacg ggcgtagatc tcgatcctct
      601 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
      661 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt
      721 tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc
      781 atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
      841 taatgcagga gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggtgcaaa
      901 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattgagggt ggtgaatgtg
      961 ctccgactca tgatcatgaa gcttttacaa agggagaccc tggacctgca ccgcgaggtc
     1021 gaggccctga tgcccgtgat gagtgagacg ctgacccggc ggggctatct ggaggtgctg
     1081 cggcgcctgc aagcggtggt cgcgccgctg gaagcacaac tcgacgccct gccgctgccg
     1141 gacgagctgg cgtggcccga gcgccgcaag gcccatctgc tcgctctgga ccttgccgta
     1201 atgggggaag ggccgtcaaa tcagcaggcg ccactgtctg tgcccgagct gaccactctg
     1261 gcgcaggcct tcggagcgct ttacgtgctt gaaggcgcga cgctaggcgg acagctcatc
     1321 agccgccatc tcaggcgcac gctggggcta agccctgagc agggcagcgc ctatttctcc
     1381 ggctatggcc cccagaccgg cccgcgctgg cgctcgttcg gggaagtgct ggaagccagc
     1441 gtgccggcag aagacgcggc agaggtcgtg gcgggagcgc ggcagacctt cggggccttc
     1501 cgccgcgcct tgcaggggct gtcggaagct gaagcggtcc aagtgcctga ggtcgcccac
     1561 gcatgacgag catcaccccg gttaccctgg cgaactgcga ggacgaaccc atccacgtgc
     1621 ctggcgcgat ccagccgcac ggggcgctgg tcaccctgcg cgcggacggc atggtcctcg
     1681 cggcaagcga gaacatccag gcgctactgg ggttcgtcgc ctcgccgggt tcctacctga
     1741 cgcaggagca ggtggggccg gaggtcctgc gcatgctcga agagggactg accggcaacg
     1801 gcccatggtc gaacagcgtc gagacgcgga tcggcgaaca cctgttcgac gtgatcggcc
     1861 acagctacaa ggaagtgttc tacctggagt tcgagatccg caccgcggac acgctgtcga
     1921 tcacctcctt caccctcaac gcccagcgca tcatcgccca ggtccagttg cacaacgaca
     1981 ccgccagcct gctcagcaac gtcaccgacg aactgcgccg gatgaccggc tacgaccggg
     2041 tgatggccta ccgtttccgc catgacgact ccggcgaggt ggtcgccgag agccgtcgcg
     2101 aagacctcga aagctacctg gggcaacgct acccggcctc ggacatcccg gcccaggcgc
     2161 gtcgcctgta catccagaac ccgatcaggc tgatcgccga cgtggcctat acgccgatgc
     2221 gcgtgtttcc cgcgctgaac ccggagacca acgaatcgtt cgatctcagc tacagcgtgc
     2281 tgcgcagcgt ttcgccgatc cactgcgaat acctgaccaa catgggcgtg cgcgcctcga
     2341 tgagcatctc catcgtggtc ggcggcaagc tctggggact gttctcctgc catcacatgt
     2401 cgcccaaact gattccctac ccggtgcgca tgtcgttcca gatcttctcc caggtctgca
     2461 gcgccatcgt cgagcgcctg gagcaggggc gtatcgccga actgctgcgc gtgtccaccg
     2521 agcgcaggct ggccctggct cgccgcgcgc gggacgccga cgacctgttc ggcgcactgg
     2581 cccacccgga cgatggcatc gccgcactga ttccctgcga cggcgccctg gtcatgctcg
     2641 gcggccgcac cctgagcatc cgcggcgact tcgagcgcca ggccgggaat gtcctgcaac
     2701 gcctgcagcg cgacccggaa cgggacatct accacaccga caactggccg cagcccagcg
     2761 aggactctcc ggacggcggc gactgctgcg gtgtgctggc gatccgtttc caccgccagg
     2821 aatcgggctg gatcttctgg ttccgccacg aggaagtgca ccgcatccgc tggggcggca
     2881 agccggaaaa gctgttgacc atcggcccct ccggcccgcg gctgaccccg cgcggctcgt
     2941 tcgaggcctg ggaggaagtg gtccgcggcc actccacgcc ctggagcgaa accgacctgg
     3001 cgatcgccga aaagctgcgc ctggacctga tggaactgtg cctcaaccat gcggcggaag
     3061 ttgatcgtat gcgtatggcg tccgcgctcg cgcacgagct caaccagccg ctggcggcga
     3121 tcagcaacta catgaagggc tcgcggcggc tgcttgccgg cagcagtgat ccgaacacac
     3181 cgaaggtcga aagcgccctg gaccgcgccg ccgagcaggc gctgcgcgcc ggccagatca
     3241 tccggcgcct gcgcgacttc gttgcccgcg gcgaatcgga gaagcgggtc gagagtctct
     3301 ccaagctgat cgaggaggcc ggcgcgctcg ggcttgccgg cgcgcgcgag cagaacgtgc
     3361 agctccgctt cagtctcgat ccgggcgccg atctcgttct cgccgaccgg gtgcagatcc
     3421 agcaggtcct ggtcaacctg ttccgcaacg cgctggaagc gatggctcag tcgcagcgac
     3481 gcgagctcgt cgtcaccaac acccccgccg ccgacgacat gatcgaggtc gaagtgtccg
     3541 acaccggcag cggtttccag gacgacgtca ttccgaacct gtttcagact ttcttcacca
     3601 ccaaggacac cggcatgggc gtgggactgt ccatcagccg ctcgatcatc gaagctcacg
     3661 gcgggcgcat gtgggccgag agcaacgcat cgggcggggc gaccttccgc ttcaccctcc
     3721 cggcagccga cgagatgata ggaggtctag catgacgacc aagggacata tctacgtcat
     3781 cgacgacgac gcggcgatgc gggattcgct gaatttcctg ctggattctg ccggcttcgg
     3841 cgtcacgctg tttgacgacg cgcaagcctt tctcgacgcc ctgccgggtc tctccttcgg
     3901 ctgcgtcgtc tccgacgtgc gcatgccggg ccttgacggc atcgagctgt tgaagcggat
     3961 gaaggcgcag caaagcccct ttccgatcct catcatgacc ggtcacggcg acgtgccgct
     4021 cgcggtcgag gcgatgaagt taggggcggt cgactttctg gaaaagcctt tcgaggacga
     4081 ccgcctcacc gccatgatcg aatcggcgat ccgccaggcc gagccggccg ccaagagcga
     4141 ggccgtcgcg caggatatcg ccgcccgcgt cgcctcgttg agccccaggg agcgccaggt
     4201 catggaaggg ctgatcgccg gcctttccaa caagctgatc gcccgcgagt acgacatcag
     4261 cccgcgcacc atcgaggtgt atcgggccaa cgtcatgacc aagatgcagg ccaacagcct
     4321 ttcggagctg gttcgcctcg cgatgcgcgc cggcatgctc aacgattgac aattgatgta
     4381 agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca
     4441 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     4501 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     4561 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     4621 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     4681 attatcgccg gcatggcggc cccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
     4741 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
     4801 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
     4861 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
     4921 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     4981 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
     5041 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
     5101 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
     5161 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
     5221 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
     5281 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
     5341 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     5401 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     5461 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     5521 catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc acagatgcgt
     5581 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6421 cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct tacataaaca
     6481 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct aggccgcgat
     6541 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     6601 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     6661 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     6721 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     6781 ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg
     6841 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     6901 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     6961 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     7021 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     7081 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     7141 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     7201 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     7261 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taagaattaa
     7321 ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7381 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     7441 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     7501 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     7561 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     7621 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     7681 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     7741 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     7801 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     7861 cccattcgcc a
//
