LOCUS       sequence-177080-        5957 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses NES-CRY2PHR-TevC of BLITz system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  AUTHORS   Lee D, Creed M, Jung K, Stefanelli T, Wendler DJ, Oh WC, Mignocchi 
            NL, Luscher C, Kwon HB
  TITLE     Temporally precise labeling and control of neuromodulatory circuits 
            in the mammalian brain.
  JOURNAL   Nat Methods. 2017 May;14(5):495-503. doi: 10.1038/nmeth.4234. Epub 
            2017 Apr 3.
  PUBMED    28369042
REFERENCE   2  (bases 1 to 5957)
  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: 
            "10.1038/nmeth.4234"; journalName: "Nat Methods"; date: "2017-05"; 
            volume: "14"; issue: "5"; pages: "495-503"
COMMENT     Sequence Label: pCMV-NES-CRY2PHR-TevC
FEATURES             Location/Qualifiers
     source          1..5957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1072..1098
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1108..2601
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     primer_bind     complement(2964..2983)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2965..2983)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2988..3122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3012..3031)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3066..3085
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3235..3255)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3235..3253)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3274..3290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3274..3290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3287..3309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3298..3314
                     /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)."
     promoter        complement(join(3322..3328,3329..3346,3347..3352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3367..3388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3505..3522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3675..4263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3755..3774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4434..5225,5226..5294))
                     /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     5057..5076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5295..5399)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5425..5880)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5562..5583)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5774..5793
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccatgcttca acttcctcct
     1081 cttgaacgtc ttactctcga ggccaccatg aagatggaca aaaagactat agtttggttt
     1141 agaagagacc taaggattga ggataatcct gcattagcag cagctgctca cgaaggatct
     1201 gtttttcctg tcttcatttg gtgtcctgaa gaagaaggac agttttatcc tggaagagct
     1261 tcaagatggt ggatgaaaca atcacttgct cacttatctc aatccttgaa ggctcttgga
     1321 tctgacctca ctttaatcaa aacccacaac acgatttcag cgatcttgga ttgtatccgc
     1381 gttaccggtg ctacaaaagt cgtctttaac cacctctatg atcctgtttc gttagttcgg
     1441 gaccataccg taaaggagaa gctggtggaa cgtgggatct ctgtgcaaag ctacaatgga
     1501 gatctattgt atgaaccgtg ggagatatac tgcgaaaagg gcaaaccttt tacgagtttc
     1561 aattcttact ggaagaaatg cttagatatg tcgattgaat ccgttatgct tcctcctcct
     1621 tggcggttga tgccaataac tgcagcggct gaagcgattt gggcgtgttc gattgaagaa
     1681 ctagggctgg agaatgaggc cgagaaaccg agcaatgcgt tgttaactag agcttggtct
     1741 ccaggatgga gcaatgctga taagttacta aatgagttca tcgagaagca gttgatagat
     1801 tatgcaaaga acagcaagaa agttgttggg aattctactt cactactttc tccgtatctc
     1861 catttcgggg aaataagcgt cagacacgtt ttccagtgtg cccggatgaa acaaattata
     1921 tgggcaagag ataagaacag tgaaggagaa gaaagtgcag atctttttct taggggaatc
     1981 ggtttaagag agtattctcg gtatatatgt ttcaacttcc cgtttactca cgagcaatcg
     2041 ttgttgagtc atcttcggtt tttcccttgg gatgctgatg ttgataagtt caaggcctgg
     2101 agacaaggca ggaccggtta tccgttggtg gatgccggaa tgagagagct ttgggctacc
     2161 ggatggatgc ataacagaat aagagtgatt gtttcaagct ttgctgtgaa gtttcttctc
     2221 cttccatgga aatggggaat gaagtatttc tgggatacac ttttggatgc tgatttggaa
     2281 tgtgacatcc ttggctggca gtatatctct gggagtatcc ccgatggcca cgagcttgat
     2341 cgcttggaca atcccgcgtt acaaggcgcc aaatatgacc cagaaggtga gtacataagg
     2401 caatggcttc ccgagcttgc gagattgcca actgaatgga tccatcatcc atgggacgct
     2461 cctttaaccg tactcaaagc ttctggtgtg gaactcggaa caaactatgc gaaacccatt
     2521 gtagacatcg acacagctcg tgagctacta gctaaagcta tttcaagaac ccgtgaagca
     2581 cagatcatga tcggagcagc actgcaggag ctcggtggag gcggccgctc cggtggcggt
     2641 ggctctacta agagcatgtc cagcatggtg agcgatacta gctgtacctt cccatcatct
     2701 gacggaatct tctggaagca ctggattcag actaaggacg gccagtgtgg cagcccactg
     2761 gtgagcacac gagacggatt catcgtgggg attcacagcg cctccaactt tacaaacacc
     2821 aataactatt tcacctcagt gccaaagaac tttatggagc tgctgaccaa ccaggaggcc
     2881 cagcagtggg tgagcgggtg gcgcctgaac gccgattccg tgctgtgggg cgggcacaag
     2941 gtgtttatgg tgtgagctag cctccctata gtgagtcgta ttacgtagat ccagacatga
     3001 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3061 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3121 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     3181 tttaattcgc gtccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     3241 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3301 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     3361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3481 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3541 gcgggagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3601 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3661 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3721 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3781 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3841 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3901 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3961 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4021 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4081 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4141 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4201 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4261 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4321 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4381 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4441 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4501 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4561 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4621 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4681 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4741 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4801 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4861 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4921 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4981 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5041 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5101 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5161 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5221 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5281 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5341 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5401 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     5461 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5521 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     5581 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     5641 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     5701 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     5761 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     5821 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     5881 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     5941 tattacgcca gtcgacc
//
