LOCUS       sequence-200870-        5481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of CIB1 (1-170) with GFP and Sec61B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5481)
  AUTHORS   Duan L, Hope J, Ong Q, Lou HY, Kim N, McCarthy C, Acero V, Lin MZ, 
            Cui B
  TITLE     Understanding CRY2 interactions for optical control of intracellular
            signaling.
  JOURNAL   Nat Commun. 2017 Sep 15;8(1):547. doi: 10.1038/s41467-017-00648-8.
  PUBMED    28916751
REFERENCE   2  (bases 1 to 5481)
  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: "Nat 
            Commun."; date: "2017-09-15"; volume: "8(1):547. doi"; pages: " 
            10.1038/s41467-017-00648-8"
COMMENT     Sequence Label: CIBN-GFP-Sec61B
FEATURES             Location/Qualifiers
     source          1..5481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             598..1107
                     /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
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     regulatory      1120..1129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1126..1128,1129..1131,1132..1842)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1126..1128,1129..1131,1132..1842)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1171..1192)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1432..1451)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1779..1800
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1855..2142
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="PGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAGR
                     TTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     polyA_signal    2264..2385
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2301..2320)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2355..2374
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2392..2847)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2529..2550)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2741..2760
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2874..2978
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2980..3337
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2980..3171
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3003..3023)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3188..3323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3372..4166
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3372..4166
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3426..3445)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4036..4055
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4354..4373)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4398..4445
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4774..5362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5263..5282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgccaccatg
      601 aatggagcta taggaggtga ccttttgctc aattttcctg acatgtcggt cctagagcgc
      661 caaagggctc acctcaagta cctcaatccc acctttgatt ctcctctcgc cggcttcttt
      721 gccgattctt caatgattac cggcggcgag atggacagct atctttcgac tgccggtttg
      781 aatcttccga tgatgtacgg tgagacgacg gtggaaggtg attcaagact ctcaatttcg
      841 ccggaaacga cgcttgggac tggaaatttc aaggcagcga agtttgatac agagactaag
      901 gattgtaatg aggcggcgaa gaagatgacg atgaacagag atgacctagt agaagaagga
      961 gaagaagaga agtcgaaaat aacagagcaa aacaatggga gcacaaaaag catcaagaag
     1021 atgaaacaca aagccaagaa agaagagaac aatttctcta atgattcatc taaagtgacg
     1081 aaggaattgg agaaaacgga ttatattcat gtaccggtcg ccaccatggt gagcaagggc
     1141 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1201 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1261 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1321 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1381 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1441 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1501 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1561 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1621 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1681 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1741 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1801 accgccgccg ggatcactct cggcatggac gagctgtaca agaacagccg cgtgcctggt
     1861 ccgaccccca gtggcactaa cgtgggatcc tcagggcgct ctcccagcaa agcagtggcc
     1921 gcccgggcgg cgggatccac tgtccggcag aggaaaaatg ccagctgtgg gacaaggagt
     1981 gcaggccgca caacctcggc aggcaccggg gggatgtggc gattctacac agaagattca
     2041 cctgggctca aagttggccc tgttccagta ttggttatga gtcttctgtt catcgcttct
     2101 gtatttatgt tgcacatttg gggcaagtac actcgttcgt aggcggccgc gactctagat
     2161 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2221 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2281 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2341 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     2401 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2461 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2521 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2581 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     2641 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     2701 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     2761 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     2821 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2881 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2941 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3001 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3061 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3121 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3181 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3241 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3301 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3361 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3421 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3481 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3541 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3601 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3661 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3721 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3781 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3841 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     3901 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     3961 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4021 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4081 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4141 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4201 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4261 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4321 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4381 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4441 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4501 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4561 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4681 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4741 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4801 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4861 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4921 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4981 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5041 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5101 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5161 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5221 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5281 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5341 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5401 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5461 tattaccgcc atgcattagt t
//
