LOCUS       sequence-376575-        9077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For 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 9077)
  AUTHORS   Multamaki E, Garcia de Fuentes A, Sieryi O, Bykov A, Gerken U, 
            Ranzani AT, Kohler J, Meglinski I, Moglich A, Takala H
  TITLE     Optogenetic Control of Bacterial Expression by Red Light.
  JOURNAL   ACS Synth Biol. 2022 Aug 23. doi: 10.1021/acssynbio.2c00259.
  PUBMED    35998606
REFERENCE   2  (bases 1 to 9077)
  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. 2022 Aug 23. doi: 10.1021/acssynbio.2c00259."
COMMENT     Sequence Label: pREDawn-AmpR-MCS
FEATURES             Location/Qualifiers
     source          1..9077
                     /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)"
     terminator      complement(402..429)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(445..516)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(564..596)
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     CDS             complement(597..1307)
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             1314..1325
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1648..1667)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1873..1950
                     /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             6226..6417
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6411..6433)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6519..6658
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6574..6593
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6673..6690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6844..7432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6924..6943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7554..7658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7659..7727,7728..8519)
                     /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(7877..7896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(8611..9066)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8748..8769)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8960..8979
                     /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 tttctagagc aaccattatc accgccagag
      361 gtaaaatagt caacacgcac ggtgttactc tagtatataa acgcagaaag gcccacccga
      421 aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa
      481 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggctctag tagcgatcta
      541 cactagcact atcagcgtta ttaagctact aaagcgtagt tttcgtcgtt tgcagcgcca
      601 aacgtctctt caggccactg actagcgata actttcccca caacggaaca actctcattg
      661 catgggatca ttgggtactg tgggtttagt ggttgtaaaa acacctgacc gctatccctg
      721 atcagtttct tgaaggtaaa ctcatcaccc ccaagtctgg ctatgcagaa atcacctggc
      781 tcaacagcct gctcagggtc aacgagaatt aacattccgt caggaaagct tggcttggag
      841 cctgttggtg cggtcatgga attaccttca acctcaagcc agaatgcaga atcactggct
      901 tttttggttg tgcttaccca tctctccgca tcacctttgg taaaggttct aagctcaggt
      961 gagaacatcc ctgcctgaac atgagaaaaa acagggtact catactcact tctaagtgac
     1021 ggctgcatac taaccgcttc atacatctcg tagatttctc tggcgattga agggctaaat
     1081 tcttcaacgc taactttgag aatttttgca agcaatgcgg cgttataagc atttaatgca
     1141 ttgatgccat taaataaagc accaacgcct gactgcccca tccccatctt gtctgcgaca
     1201 gattcctggg ataagccaag ttcatttttc tttttttcat aaattgcttt aaggcgacgt
     1261 gcgtcctcaa gctgctcttg tgttaatggt ttcttttttg tgctcatcta gtatttctcc
     1321 tcttttctag actccgttgt gatgacgcat tggtacgcgg tatcgggagg ttcgaaaatt
     1381 tcgagcgata tcttaagggg ggtgccttac gtagaacccc gtaggtcatg cccgaggccg
     1441 gtcctggatg gcgcggcgga tacgcttgag caggttttcg tcgagaagcg gcttcaaaac
     1501 cacgtctttt acgccggcct cggcggcccg ggtcgagatg ttttcgtccg gatagccggt
     1561 gatcaggatc acgggcgtag atctcgatcc tctacgccgg acgcatcgtg gccggcatca
     1621 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     1681 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
     1741 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
     1801 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
     1861 gtcgagatcc cggacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc
     1921 ccggaagaga gtcaattgag ggtggtgaat gtgctccgac tcatgatcat gaagctttta
     1981 caaagggaga ccctggacct gcaccgcgag gtcgaggccc tgatgcccgt gatgagtgag
     2041 acgctgaccc ggcggggcta tctggaggtg ctgcggcgcc tgcaagcggt ggtcgcgccg
     2101 ctggaagcac aactcgacgc cctgccgctg ccggacgagc tggcgtggcc cgagcgccgc
     2161 aaggcccatc tgctcgctct ggaccttgcc gtaatggggg aagggccgtc aaatcagcag
     2221 gcgccactgt ctgtgcccga gctgaccact ctggcgcagg ccttcggagc gctttacgtg
     2281 cttgaaggcg cgacgctagg cggacagctc atcagccgcc atctcaggcg cacgctgggg
     2341 ctaagccctg agcagggcag cgcctatttc tccggctatg gcccccagac cggcccgcgc
     2401 tggcgctcgt tcggggaagt gctggaagcc agcgtgccgg cagaagacgc ggcagaggtc
     2461 gtggcgggag cgcggcagac cttcggggcc ttccgccgcg ccttgcaggg gctgtcggaa
     2521 gctgaagcgg tccaagtgcc tgaggtcgcc cacgcatgag ccgggacccg ttgccctttt
     2581 ttccaccgct ttaccttggt ggcccggaaa ttaccaccga gaactgcgag cgcgagccga
     2641 ttcatattcc cggcagcatc cagccgcacg gcgccctgct cactgccgac gggcacagcg
     2701 gcgaggtgct ccagatgagc ctcaacgcgg ccacttttct gggacaggaa cccacagtgc
     2761 tgcgcggaca gaccctcgcc gcactgctgc ccgagcagtg gcccgcgctg caagcggccc
     2821 tgccccccgg ctgccccgac gccctgcaat accgcgcaac gctggactgg cctgccgccg
     2881 ggcacctttc gctgacggtg caccgggtcg gcgagttgct gattctggaa ttcgagccga
     2941 cggaggcctg ggacagcacc gggccgcacg cgctgcgcaa cgcgatgttc gcgctcgaaa
     3001 gtgcccccaa cctgcgggcg ctggccgagg tggcgaccca gacggtccgc gagctgacgg
     3061 gctttgaccg ggtgatgctc tacaaatttg cccccgacgc caccggcgaa gtgattgccg
     3121 aggcccgccg tgaggggctg cacgcctttc tgggccaccg ttttcccgcg tcggacattc
     3181 cggcgcaggc ccgcgcgctc tacacccggc acctgctgcg cctgaccgcc gacacccgcg
     3241 ccgccgccgt gccgctcgat cccgtcctca acccgcagac gaatgcgccc accccgctgg
     3301 gcggcgccgt gctgcgcgcc acctcgccca tgcacatgca gtacctgcgg aacatgggcg
     3361 tcgggtcgag cctgtcggtg tcggtggtgg tcggcggcca gctctggggc ctgatcgcct
     3421 gccaccacca gacgccctac gtgttgccgc ccgacctgcg aaccacgctc gaatacctgg
     3481 gccgcttgct gagcctgcaa gttcaggtca aggaagcggc ggacgtggcg gcctttcgcc
     3541 agagcctgcg ggagcaccac gcgcgggtgg ccctcgcggc ggcgcactcg ctctcgccgc
     3601 acgacaccct cagtgacccg gcgcttgacc tgctgggcct gatgcgggcc gggggcctga
     3661 ttctgcgttt cgagggccgc tggcagacgt tgggtgaagt gccgcctgcc ccggcggtgg
     3721 acgcgctgct ggcgtggctc gaaacccagc cgggcgccct ggtccagacc gacgcgctag
     3781 gccaactgtg gcccgccggc gccgatctcg cccccagcgc agcgggcctg ctcgccatca
     3841 gcgtgggcga gggctggtcg gagtgcctcg tctggctgcg gcccgaactg cggctggagg
     3901 tcgcctgggg cggggccact cctgaccagg cgaaagacga cctcgggccg cgccactcat
     3961 tcgacaccta cctcgaagaa aaacgcggct acgccgagcc ctggcatccc ggcgaaatcg
     4021 aggaggcgca ggatctacgt gacacattga ccggggcgct gggcgagcgc ctgctccagg
     4081 aactgcaatc cgagctcgtc cacgtctcca ggctgagcgc catgggcgaa atggcgtccg
     4141 cgctcgcgca cgagctcaac cagccgctgg cggcgatcag caactacatg aagggctcgc
     4201 ggcggctgct tgccggcagc agtgatccga acacaccgaa ggtcgaaagc gccctggacc
     4261 gcgccgccga gcaggcgctg cgcgccggcc agatcatccg gcgcctgcgc gacttcgttg
     4321 cccgcggcga atcggagaag cgggtcgaga gtctctccaa gctgatcgag gaggccggcg
     4381 cgctcgggct tgccggcgcg cgcgagcaga acgtgcagct ccgcttcagt ctcgatccgg
     4441 gcgccgatct cgttctcgcc gaccgggtgc agatccagca ggtcctggtc aacctgttcc
     4501 gcaacgcgct ggaagcgatg gctcagtcgc agcgacgcga gctcgtcgtc accaacaccc
     4561 ccgccgccga cgacatgatc gaggtcgaag tgtccgacac cggcagcggt ttccaggacg
     4621 acgtcattcc gaacctgttt cagactttct tcaccaccaa ggacaccggc atgggcgtgg
     4681 gactgtccat cagccgctcg atcatcgaag ctcacggcgg gcgcatgtgg gccgagagca
     4741 acgcatcggg cggggcgacc ttccgcttca ccctcccggc agccgacgag atgataggag
     4801 gtctagcatg acgaccaagg gacatatcta cgtcatcgac gacgacgcgg cgatgcggga
     4861 ttcgctgaat ttcctgctgg attctgccgg cttcggcgtc acgctgtttg acgacgcgca
     4921 agcctttctc gacgccctgc cgggtctctc cttcggctgc gtcgtctccg acgtgcgcat
     4981 gccgggcctt gacggcatcg agctgttgaa gcggatgaag gcgcagcaaa gcccctttcc
     5041 gatcctcatc atgaccggtc acggcgacgt gccgctcgcg gtcgaggcga tgaagttagg
     5101 ggcggtcgac tttctggaaa agcctttcga ggacgaccgc ctcaccgcca tgatcgaatc
     5161 ggcgatccgc caggccgagc cggccgccaa gagcgaggcc gtcgcgcagg atatcgccgc
     5221 ccgcgtcgcc tcgttgagcc ccagggagcg ccaggtcatg gaagggctga tcgccggcct
     5281 ttccaacaag ctgatcgccc gcgagtacga catcagcccg cgcaccatcg aggtgtatcg
     5341 ggccaacgtc atgaccaaga tgcaggccaa cagcctttcg gagctggttc gcctcgcgat
     5401 gcgcgccggc atgctcaacg attgacaatt gatgtaagtt agctcactca ttaggcaccg
     5461 ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     5521 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     5581 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     5641 atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     5701 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggcccca
     5761 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     5821 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     5881 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     5941 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     6001 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     6061 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     6121 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     6181 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     6241 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     6301 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     6361 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6421 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6481 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     6541 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     6601 tgcaccattg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     6661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7261 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7501 ggtcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttcgcggaa
     7561 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     7621 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     7681 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     7741 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     7801 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     7861 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     7921 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     7981 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     8041 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     8101 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     8161 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     8221 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     8281 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     8341 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     8401 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     8461 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaag
     8521 aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8581 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     8641 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     8701 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     8761 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     8821 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     8881 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     8941 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     9001 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     9061 gcgcgtccca ttcgcca
//
