LOCUS       sequence-468089-        7466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For far-red light-induced bacterial expression of gene of interest 
            (AmpR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7466)
  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 7466)
  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-AmpR-MCS
FEATURES             Location/Qualifiers
     source          1..7466
                     /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."
     rep_origin      5170..5908
                     /direction=RIGHT
                     /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."
     promoter        5956..6047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6048..6116,6117..6908)
                     /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(6266..6285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7000..7455)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7137..7158)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7349..7368
                     /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 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tcttttgccc
     5221 tgtaaacgaa aaaaccacct ggggaggtgg tttgatcgaa ggttaagtca gttggggaac
     5281 tgcttaaccg tggtaactgg ctttcgcaga gcacagcaac caaatctgtc cttccagtgt
     5341 agccggactt tggcgcacac ttcaagagca accgcgtgtt tagctaaaca aatcctctgc
     5401 gaactcccag ttaccaatgg ctgctgccag tggcgtttta ccgtgctttt ccgggttgga
     5461 ctcaagtgaa cagttaccgg ataaggcgca gcagtcgggc tgaacgggga gttcttgctt
     5521 acagcccagc ttggagcgaa cgacctacac cgagccgaga taccagtgtg tgagctatga
     5581 gaaagcgcca cacttcccgt aagggagaaa ggcggaacag gtatccggta aacggcaggg
     5641 tcggaacagg agagcgcaag agggagcgac ccgccggaaa cggtggggat ctttaagtcc
     5701 tgtcgggttt cgcccgtact gtcagattca tggttgagcc tcacggctcc cacagatgca
     5761 ccggaaaagc gtctgtttat gtgaactctg gcaggagggc ggagcctatg gaaaaacgcc
     5821 accggcgcgg ccctgctgtt ttgcctcaca tgttagtccc ctgcttatcc acggaatctg
     5881 tgggtaactt tgtatgtgtc cgcagcgccc gccgcagtct cacgcccgga gcgtagcgac
     5941 cgagtgagct agctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6001 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6061 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6121 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6181 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6241 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6301 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6361 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6421 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6481 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6541 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     6601 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6661 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6721 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6781 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6841 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6901 attggtaaga attaattcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6961 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     7021 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     7081 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     7141 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     7201 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     7261 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     7321 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     7381 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     7441 cgctacaggg cgcgtcccat tcgcca
//
