LOCUS       sequence-219934-        8959 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 8959)
  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 8959)
  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..8959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1377..1886
                     /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             2400..2420
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2425..2441)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2425..2441)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3550..3569)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3551..3569)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3576..3593)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3576..3592)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3585..3607)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3733..4188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3820..3839)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4030..4051
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4544..4563)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4663..4685
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4723..4741)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4855..4959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4960..5028,5029..5820)
                     /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(5178..5197)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5991..6579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6480..6499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6733..6750
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6867..6888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6903..6908,6909..6926,6927..6933)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6941..6957
                     /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     6946..6968
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6965..6981
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6965..6981
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7002..7020
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7346..7533
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 aaggtgagac gcgcataacc gctagagtac tttgaagagg aaacagcaat agggttgcta
       61 ccagtataaa tagacaggta catacaacac tggaaatggt tgtctgtttg agtacgcttt
      121 caattcattt gggtgtgcac tttattatgt tacaatatgg aagggaactt tacacttctc
      181 ctatgcacat atattaatta aagtccaatg ctagtagaga aggggggtaa cacccctccg
      241 cgctcttttc cgattttttt ctaaaccgtg gaatatttcg gatatccttt tgttgtttcc
      301 gggtgtacaa tatggacttc ctcttttctg gcaaccaaac ccatacatcg ggattcctat
      361 aataccttcg ttggtctccc taacatgtag gtggcggagg ggagatatac aatagaacag
      421 ataccagaca agacataatg ggctaaacaa gactacacca attacactgc ctcattgatg
      481 gtggtacata acgaactaat actgtagccc tagacttgat agccatcatc atatcgaagt
      541 ttcactaccc tttttccatt tgccatctat tgaagtaata ataggcgcat gcaacttctt
      601 ttcttttttt ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa
      661 aaaatgatgg aagacactaa aggaaaaaat taacgacaaa gacagcacca acagatgtcg
      721 ttgttccaga gctgatgagg ggtatctcga agcacacgaa actttttcct tccttcattc
      781 acgcacacta ctctctaatg agcaacggta tacggccttc cttccagtta cttgaatttg
      841 aaataaaaaa aagtttgctg tcttgctatc aagtataaat agacctgcaa ttattaatct
      901 tttgtttcct cgtcattgtt ctcgttccct ttcttccttg tttctttttc tgcacaatat
      961 ttcaagctat accaagcata caactcgaga ataaaatgtt atctcctttt accaataacc
     1021 accatgccag ttcggggcaa aataatagcg agaatagtga ggaaagaatt atagatatta
     1081 gatctgtccc aaatgctttg atgaatgaac aaaatagtaa ggctgaattg cacaattggg
     1141 agccaccgtt atatgattcg gtcggtaacc acaatcatgc tggcactatg gagagtcatc
     1201 cacatacaaa cattcatgaa gaattaaagg agttcttatc aggcgatttg attgaagccg
     1261 aagataatgc aaaattaatg tttggagatg acaactctaa tccagctgac tatcttttag
     1321 aatttggttc ggggggttct ggatctagtt caggatctgg ttcagaatct ggttcaatga
     1381 atggagctat aggaggtgac cttttgctca attttcctga catgtcggtc ctagagcgcc
     1441 aaagggctca cctcaagtac ctcaatccca cctttgattc tcctctcgcc ggcttctttg
     1501 ccgattcttc aatgattacc ggcggcgaga tggacagcta tctttcgact gccggtttga
     1561 atcttccgat gatgtacggt gagacgacgg tggaaggtga ttcaagactc tcaatttcgc
     1621 cggaaacgac gcttgggact ggaaatttca agaaacggaa gtttgataca gagactaagg
     1681 attgtaatga gaagaagaag aagatgacga tgaacagaga tgacctagta gaagaaggag
     1741 aagaagagaa gtcgaaaata acagagcaaa acaatgggag cacaaaaagc atcaagaaga
     1801 tgaaacacaa agccaagaaa gaagagaaca atttctctaa tgattcatct aaagtgacga
     1861 aggaattgga gaaaacggat tatattcatg ttcgtgcacg acgaggccaa gccactgata
     1921 gtcacagcat agcagaacga gttagaagag aaaagatcag tgagagaatg aagtttctac
     1981 aagatttggt tcctggatgc gacaagatca caggcaaagc agggatgctt gatgaaatca
     2041 ttaactatgt tcagtctctt cagagacaaa tcgagttctt atcgatgaaa ctagcaattg
     2101 tgaatccaag gccggatttt gatatggatg acatttttgc caaagaggtt gcctcaactc
     2161 caatgactgt ggtgccatct cctgaaatgg ttctttccgg ttattctcat gagatggttc
     2221 actctggtta ttctagtgag atggttaact ccggttacct tcatgtcaat ccaatgcagc
     2281 aagtgaatac cagttctgat ccattgtcat gcttcaacaa tggcgaagct ccttcgatgt
     2341 gggactctca tgtgcagaat ctctatggca atttaggagt tggttctggt tctggttctc
     2401 ccaagaagaa gcggaaggtc taaggatcca ctagttctag acacctgcaa catgcgtgag
     2461 taaataagcg gccgccaccg cggtggagct ctaagcaaat agctaaatta tatacgaatt
     2521 aatattatga ttaagtgttt acgtgagtgc gatattttta ttactatctt atacagttgt
     2581 atatactcta taaaatgagt tgtctattaa ttaacgcgat gaatgctttc tgggtttacc
     2641 tctccaacaa ctctagttta cttctcaata cattcaattg tatttgattt gtcaatactt
     2701 catcattaat caattctata gttttgtttt tctcgtttat ttccaaattt aatgcatcaa
     2761 ttttattatt caatttgtcg ttgattttgg ttaatgattt tatggtttga tctctggcat
     2821 tgattgtttg tgttagtttt tcattattga taattaaatt atttaagtta gttatcaact
     2881 cggtgttttc aagtttcaag ttttcaattt ctttagagtt tattagattt gtcaaagttt
     2941 ctgaattgct tgattggtcc tgtagaagag tatttgttgt tgtggataat tgattcaatt
     3001 tttgagacaa ttgctggaag gcgttgaaat atctagcatc aatctcatgg tttttttccc
     3061 gagagtctcg tagattcaat tgttttaata tatcttggga ccactcttga tttgaactca
     3121 tggaaattaa actgggtgtt gtgttgtggt gtaatgattg tacccccttt gcttataatt
     3181 gtgtgggaga tggaggaacg ggaaaaagtt agttgtggtg ataggtggca agtggtattc
     3241 cgtaagaaca acaagaaaag catttcatat tatggctgaa ctgagcgaac aagtgcaaaa
     3301 tttaagcatc aacgacaaca acgagaatgg ttatgttcct cctcacttaa gaggaaaacc
     3361 aagaagtgcc agaaataaca gtagcaacta caataacaac aacggcggct acaacggtgg
     3421 ccgtggcggt ggcagcttct ttagcaacaa ccgtcgtggt ggttacggca acggtggttt
     3481 cttcggtgga aacaacggtg gcagcggccg gccatttaaa tgtttaaacg gccacgctgg
     3541 cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
     3601 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     3661 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3721 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3781 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3841 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3901 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3961 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4021 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4081 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4141 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     4201 ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt
     4261 aaattgaagc tctaatttgt gagtttagta ttaggatctc tacccttggc gaaaagtcct
     4321 ctgccaacaa tgatgatatc tgatccacca cttacaactt cgtcgacggt tctgtactgc
     4381 tgacccaata tgcgtatata taccaatcta agtctgtgct ccttccttcg ttcttccttc
     4441 tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa aaaagaataa
     4501 aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct cagtacaatc tgctctgatg
     4561 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     4621 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     4681 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     4741 ttttataggt taatgtcatg ataataatgg tttcttagac gtcggcgcgc cctgatccgt
     4801 cctaagaaac caggcgcgcc gaggacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4861 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4921 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4981 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5041 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5101 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5161 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5221 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5281 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5341 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5401 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5461 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5521 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5581 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5641 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5701 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5761 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5821 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5881 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5941 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6001 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6061 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6121 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6181 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6241 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6301 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6361 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6421 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6481 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6541 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6601 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6661 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6721 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6781 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6841 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     6901 gctttacact ttatgcttcc ggctcctatg ttgtgtggaa ttgtgagcgg ataacaattt
     6961 cacacaggaa acagctatga ccatgattac gccaagctcg gaattaaccc tcactaaagg
     7021 gaacaaaagc tgggccggcc atttaaatgt ttaaacggcc acgctggccg ctgcacggtc
     7081 ctgttcccta gcatgtacgt gagcgtattt ccttttaaac cacgacgctt tgtcttcatt
     7141 caacgtttcc cattgttttt ttctactatt gctttgctgt gggaaaaact tatcgaaaga
     7201 tgacgacttt ttcttaattc tcgttttaag agcttggtga gcgctaggag tcactgccag
     7261 gtatcgtttg aacacggcat tagtcaggga agtcataaca cagtcctttc ccgcaatttt
     7321 ctttttctat tactcttggc ctcctctagt acactctata tttttttatg cctcggtaat
     7381 gattttcatt tttttttttc cacctagcgg atgactcttt ttttttctta gcgattggca
     7441 ttatcacata atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta
     7501 aaaaatgagc aggcaagata aacgaaggca aagaggctct atatgtatat aaagctgcgg
     7561 atgccttctg atttaatcta gttctagttt acttcaggtt tacttaaagt tgagataatt
     7621 gaagcaaaga gaagcatcga gcaaaggatg gcgtttgtta agagggttac gcaggagacg
     7681 aatatacagc tggcgctgga tcttgacggt gggtctgttt ctgtacggga gagcatactg
     7741 ggcaaggaat atgctagtgg tgatgggcag accatccatg tgcacactgg agttgggttt
     7801 ttggaccaca tgttgactgc gctggcgaag catggcgggt ggtctctgat cctggagtgt
     7861 ataggggact tgcacatcga tgaccaccac actgttgagg actgtgggat tgcgctgggc
     7921 caagcgttca aggaggcgct tggctccgtg cgtggtatca agaggttcgg gcatgggttt
     7981 gcaccactgg acgaggcgct gagccgcgct gtggttgact tctccaatag gcctttcgcc
     8041 gtggtggagc tgggcctgaa gcgagaacgc ataggccagc tatccacaga gatgatcccg
     8101 cacttcttgg agagtttcgc cactgaggcg cgtatcacca tgcatgtgga ctgtctgcgg
     8161 ggcaccaacg accaccaccg ctccgaatca gctttcaagg cgctcgccat cgccatcaga
     8221 gaggcaagaa cacctacggg tcgcgatgac gttccatcca ctaagggtgt cctagcatag
     8281 aacacagccc acagctacca ccatcaacaa tatttatata gataacgtac acatagaaat
     8341 cacacaaaca gagtatttat tcttaactac atgaactacc atcagaccgt ctgggccact
     8401 atataatgtg ctattcataa acgtgatcac tctacgtagc aggcgacccc aggtgaaaat
     8461 ttttcagcga gctgccagat tgtcaggtga aaaactgaaa aaaacttctg ggcgatgagc
     8521 ttgtggcggg aaattaagta tataaagcag ttagtttctt ctgcttcttg tggttctggg
     8581 tattcttgta atatttggaa taatagaaaa ccaagactac tgcctttctt ttcatattat
     8641 agaggaagaa atacgcacga acacgatata gaggtaaatt ttttgcaaaa agataccaaa
     8701 ctataaaaga aatagaagac taaactacaa aaaataaaga aagtaataca tatatccaat
     8761 atgactgagt tagcagagca agtttccatg cctgaagggc aacaaagacg tggtggttac
     8821 gttccacctc acatccgtaa cggtggtggc agaagaggcg attctgctcc tcgtggtggt
     8881 ttcaacaacg gtggcggccg tggtggtttc ttcagaaaca acggtgaccg tggtggattc
     8941 ggctttgggt accgggccc
//
