LOCUS       sequence-218857-        8929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  blue light inducible cryptochrome based optogenetic expression 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8929)
  AUTHORS   Harrigan P, Madhani HD, El-Samad H
  TITLE     Real-Time Genetic Compensation Defines the Dynamic Demands of 
            Feedback Control.
  JOURNAL   Cell. 2018 Oct 18;175(3):877-886.e10. doi: 
            10.1016/j.cell.2018.09.044.
  PUBMED    30340045
REFERENCE   2  (bases 1 to 8929)
  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"; doi: 
            "10.1016/j.cell.2018.09"; journalName: "Cell"; date: "2018-10-18- 
            18"; volume: "175"; issue: "3"; pages: "877-886.e10"
COMMENT     Sequence Label: pHES 946
FEATURES             Location/Qualifiers
     source          1..8929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1864..2373
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKKRKFDTETKDCN
                     EKKKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             2887..2907
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2912..2928)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2912..2928)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4030..4049)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4031..4049)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4056..4073)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4056..4072)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4065..4087)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4213..4668
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4300..4319)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4510..4531
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5024..5043)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5143..5165
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5203..5221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5306..5410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5411..5479,5480..6271)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5629..5648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6442..7030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6931..6950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7184..7201
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7318..7339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7354..7359,7360..7377,7378..7384)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7392..7408
                     /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)."
     primer_bind     7397..7419
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7416..7432
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7416..7432
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7453..7471
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7797..7985
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 gattgtcagg tgaaaaactg aaaaaaactt ctgggcgatg agcttgtggc gggaaattaa
       61 gtatataaag cagttagttt cttctgcttc ttgtggttct gggtattctt gtaatatttg
      121 gaataataga aaaccaagac tactgccttt cttttcatat tatagaggaa gaaatacgca
      181 cgaacacgat atagaggtaa attttttgca aaaagatacc aaactataaa agaaatagaa
      241 gactaaacta caaaaaataa agaaagtaat acatatatcc aatatgactg agttagcaga
      301 gcaagtttcc atgcctgaag ggcaacaaag acgtggtggt tacgttccac ctcacatccg
      361 taacggtggt ggcagaagag gcgattctgc tcctcgtggt ggtttcaaca acggtggcgg
      421 ccgtggtggt ttcttcagaa acaacggtga ccgtggtgga ttcggctttg ggtaccgggc
      481 ccaataaaaa ggtgagacgc gcataaccgc tagagtactt tgaagaggaa acagcaatag
      541 ggttgctacc agtataaata gacaggtaca tacaacactg gaaatggttg tctgtttgag
      601 tacgctttca attcatttgg gtgtgcactt tattatgtta caatatggaa gggaacttta
      661 cacttctcct atgcacatat attaattaaa gtccaatgct agtagagaag gggggtaaca
      721 cccctccgcg ctcttttccg atttttttct aaaccgtgga atatttcgga tatccttttg
      781 ttgtttccgg gtgtacaata tggacttcct cttttctggc aaccaaaccc atacatcggg
      841 attcctataa taccttcgtt ggtctcccta acatgtaggt ggcggagggg agatatacaa
      901 tagaacagat accagacaag acataatggg ctaaacaaga ctacaccaat tacactgcct
      961 cattgatggt ggtacataac gaactaatac tgtagcccta gacttgatag ccatcatcat
     1021 atcgaagttt cactaccctt tttccatttg ccatctattg aagtaataat aggcgcatgc
     1081 aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc atatccgcaa
     1141 tgacaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca
     1201 gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc
     1261 ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt
     1321 gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta
     1381 ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc
     1441 acaatatttc aagctatacc aagcatacaa ctcgagaata aaatgttatc tccttttacc
     1501 aataaccacc atgccagttc ggggcaaaat aatagcgaga atagtgagga aagaattata
     1561 gatattagat ctgtcccaaa tgctttgatg aatgaacaaa atagtaaggc tgaattgcac
     1621 aattgggagc caccgttata tgattcggtc ggtaaccaca atcatgctgg cactatggag
     1681 agtcatccac atacaaacat tcatgaagaa ttaaaggagt tcttatcagg cgatttgatt
     1741 gaagccgaag ataacgcaaa attaatgttt ggagatgaca actctaatcc agctgactat
     1801 cttttagaat ttggttcggg gggttctgga tctagttcag gatctggttc agaatctggt
     1861 tcaatgaatg gagctatagg aggtgacctt ttgctcaatt ttcctgacat gtcggtccta
     1921 gagcgccaaa gggctcacct caagtacctc aatcccacct ttgattctcc tctcgccggc
     1981 ttctttgccg attcttcaat gattaccggc ggcgagatgg acagctatct ttcgactgcc
     2041 ggtttgaatc ttccgatgat gtacggtgag acgacggtgg aaggtgattc aagactctca
     2101 atttcgccgg aaacgacgct tgggactgga aatttcaaga aacggaagtt tgatacagag
     2161 actaaggatt gtaatgagaa gaagaagaag atgacgatga acagagatga cctagtagaa
     2221 gaaggagaag aagagaagtc gaaaataaca gagcaaaaca atgggagcac aaaaagcatc
     2281 aagaagatga aacacaaagc caagaaagaa gagaacaatt tctctaatga ttcatctaaa
     2341 gtgacgaagg aattggagaa aacggattat attcatgttc gtgcacgacg aggccaagcc
     2401 actgatagtc acagcatagc agaacgagtt agaagagaaa agatcagtga gagaatgaag
     2461 tttctacaag atttggttcc tggatgcgac aagatcacag gcaaagcagg gatgcttgat
     2521 gaaatcatta actatgttca gtctcttcag agacaaatcg agttcttatc gatgaaacta
     2581 gcaattgtga atccaaggcc ggattttgat atggatgaca tttttgccaa agaggttgcc
     2641 tcaactccaa tgactgtggt gccatctcct gaaatggttc tttccggtta ttctcatgag
     2701 atggttcact ctggttattc tagtgagatg gttaactccg gttaccttca tgtcaatcca
     2761 atgcagcaag tgaataccag ttctgatcca ttgtcatgct tcaacaatgg cgaagctcct
     2821 tcgatgtggg actctcatgt gcagaatctc tatggcaatt taggagttgg ttctggttct
     2881 ggttctccca agaagaagcg gaaggtctaa ggatccacta gttctagaca cctgcaacat
     2941 gcgtgagtaa ataagcggcc gccaccgcgg tggagctcta agcaaatagc taaattatat
     3001 acgaattaat attatgatta agtgtttacg tgagtgcgat atttttatta ctatcttata
     3061 cagttgtata tactctataa aatgagttgt ctattaatta acgcgatgaa tgctttctgg
     3121 gtttacctct ccaacaactc tagtttactt ctcaatacat tcaattgtat ttgatttgtc
     3181 aatacttcat cattaatcaa ttctatagtt ttgtttttct cgtttatttc caaatttaat
     3241 gcatcaattt tattattcaa tttgtcgttg attttggtta atgattttat ggtttgatct
     3301 ctggcattga ttgtttgtgt tagtttttca ttattgataa ttaaattatt taagttagtt
     3361 atcaactcgg tgttttcaag tttcaagttt tcaatttctt tagagtttat tagatttgtc
     3421 aaagtttctg aattgcttga ttggtcctgt agaagagtat ttgttgttgt ggataattga
     3481 ttcaattttt gagacaattg ctggaaggcg ttgaaatatc tagcatcaat ctcatggttt
     3541 ttttcccgag agtctcgtag attcaattgt tttaatatat cttgggacca ctcttgattt
     3601 gaactcatgg aaattaaact gggtgttgtg ttgtggtgta atgattgtac cccctttgct
     3661 tataattgtg tgggagatgg aggaacggga aaaagttagt tgtggtgata ggtggcaagt
     3721 ggtattccgt aagaacaaca agaaaagcat ttcatattat ggctgaactg agcgaacaag
     3781 tgcaaaattt aagcatcaac gacaacaacg agaatggtta tgttcctcct cacttaagag
     3841 gaaaaccaag aagtgccaga aataacagta gcaactacaa taacaacaac ggcggctaca
     3901 acggtggccg tggcggtggc agcttcttta gcaacaaccg tcgtggtggt tacggcaacg
     3961 gtggtttctt cggtggaaac aacggtggca gcggccggcc atttaaatgt ttaaacggcc
     4021 acgctggccc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     4081 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     4141 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4201 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4261 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4321 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4381 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4441 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4501 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4561 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4621 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     4681 ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt
     4741 aaattgaagc tctaatttgt gagtttagta ttaggatctc tacccttggc gaaaagtcct
     4801 ctgccaacaa tgatgatatc tgatccacca cttacaactt cgtcgacggt tctgtactgc
     4861 tgacccaata tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc
     4921 tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa
     4981 aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct cagtacaatc tgctctgatg
     5041 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5101 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5161 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5221 ttttataggt taatgtcatg ataataatgg tttcttagac gtcggcgcgc cgaggacgtc
     5281 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5341 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5401 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5461 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5521 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5581 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5641 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5701 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5761 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5821 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5881 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5941 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6001 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6061 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6121 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6181 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6241 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6301 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6361 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6421 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6481 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6541 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6601 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6661 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6721 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6781 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6841 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6901 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6961 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7021 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7081 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7141 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7201 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7261 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7321 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7381 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     7441 cgccaagctc ggaattaacc ctcactaaag ggaacaaaag ctgggccggc catttaaatg
     7501 tttaaacggc cacgctggcc gctgcacggt cctgttccct agcatgtacg tgagcgtatt
     7561 tccttttaaa ccacgacgct ttgtcttcat tcaacgtttc ccattgtttt tttctactat
     7621 tgctttgctg tgggaaaaac ttatcgaaag atgacgactt tttcttaatt ctcgttttaa
     7681 gagcttggtg agcgctagga gtcactgcca ggtatcgttt gaacacggca ttagtcaggg
     7741 aagtcataac acagtccttt cccgcaattt tctttttcta ttactcttgg cctcctctag
     7801 tacactctat atttttttat gcctcggtaa tgattttcat tttttttttt tccacctagc
     7861 ggatgactct ttttttttct tagcgattgg cattatcaca taatgaatta tacattatat
     7921 aaagtaatgt gatttcttcg aagaatatac taaaaaatga gcaggcaaga taaacgaagg
     7981 caaagaggct ctatatgtat ataaagctgc ggatgccttc tgatttaatc tagttctagt
     8041 ttacttcagg tttacttaaa gttgagataa ttgaagcaaa gagaagcatc gagcaaagga
     8101 tggcgtttgt taagagggtt acgcaggaga cgaatataca gctggcgctg gatcttgacg
     8161 gtgggtctgt ttctgtacgg gagagcatac tgggcaagga atatgctagt ggtgatgggc
     8221 agaccatcca tgtgcacact ggagttgggt ttttggacca catgttgact gcgctggcga
     8281 agcatggcgg gtggtctctg atcctggagt gtatagggga cttgcacatc gatgaccacc
     8341 acactgttga ggactgtggg attgcgctgg gccaagcgtt caaggaggcg cttggctccg
     8401 tgcgtggtat caagaggttc gggcatgggt ttgcaccact ggacgaggcg ctgagccgcg
     8461 ctgtggttga cttctccaat aggcctttcg ccgtggtgga gctgggcctg aagcgagaac
     8521 gcataggcca gctatccaca gagatgatcc cgcacttctt ggagagtttc gccactgagg
     8581 cgcgtatcac catgcatgtg gactgtctgc ggggcaccaa cgaccaccac cgctccgaat
     8641 cagctttcaa ggcgctcgcc atcgccatca gagaggcaag aacacctacg ggtcgcgatg
     8701 acgttccatc cactaagggt gtcctagcat agaacacagc ccacagctac caccatcaac
     8761 aatatttata tagataacgt acacatagaa atcacacaaa cagagtattt attcttaact
     8821 acatgaacta ccatcagacc gtctgggcca ctatataatg tgctattcat aaacgtgatc
     8881 actctacgta gcaggcgacc ccaggtgaaa atttttcagc gagctgcca
//
