LOCUS       sequence-456519-        5374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of CIBN domain fused to GFP tagged mouse Syntaxin1 TMD 
            domain (transmembrane domain and polybasic region).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5374)
  AUTHORS   Idevall-Hagren O, Dickson EJ, Hille B, Toomre DK, De Camilli P
  TITLE     Optogenetic control of phosphoinositide metabolism.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):E2316-23. doi: 
            10.1073/pnas.1211305109. Epub 2012 Jul 30.
  PUBMED    22847441
REFERENCE   2  (bases 1 to 5374)
  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: 
            "10.1073/pnas.1211305109"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2012-08-28- 28"; volume: "109"; issue: "35"; pages: 
            "E2316-23"
COMMENT     Sequence Label: CIBN-GFP-TMD
FEATURES             Location/Qualifiers
     source          1..5374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(27..48)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(join(183..695,696..698,699..701))
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     376..395
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     635..656
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(717..1226)
                     /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"
     promoter        complement(1261..1464)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1290..1310)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1465..1768
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(1943..2531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2023..2042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2860..2907
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2932..2951
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3139..3933)
                     /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             complement(3139..3933)
                     /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(3250..3269)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3860..3879
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3968..4325)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3982..4117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4134..4325
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4282..4302
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(4327..4431)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4458..4913
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4545..4564)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4755..4776
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4920..5041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4931..4950)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4985..5004
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    5165..5230
                     /label=MCS
                     /note="multiple cloning site"
ORIGIN
        1 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
       61 tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc
      121 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
      181 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
      241 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
      301 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
      361 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
      421 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
      481 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
      541 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
      601 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
      661 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggcgaccgg tacatgaata
      721 taatccgttt tctccaattc cttcgtcact ttagatgaat cattagagaa attgttctct
      781 tctttcttgg ctttgtgttt catcttcttg atgctttttg tgctcccatt gttttgctct
      841 gttattttcg acttctcttc ttctccttct tctactaggt catctctgtt catcgtcatc
      901 ttcttcgccg cctcattaca atccttagtc tctgtatcaa acttcgctgc cttgaaattt
      961 ccagtcccaa gcgtcgtttc cggcgaaatt gagagtcttg aatcaccttc caccgtcgtc
     1021 tcaccgtaca tcatcggaag attcaaaccg gcagtcgaaa gatagctgtc catctcgccg
     1081 ccggtaatca ttgaagaatc ggcaaagaag ccggcgagag gagaatcaaa ggtgggattg
     1141 aggtacttga ggtgagccct ttggcgctct aggaccgaca tgtcaggaaa attgagcaaa
     1201 aggtcacctc ctatagctcc attcatggtg gcgctagcgg atctgacggt tcactaaacc
     1261 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     1321 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     1381 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     1441 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     1501 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     1561 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     1621 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     1681 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     1741 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     1801 ccccgtaatt gattactatt aataactaat gcatggcggt aatacggtta tccacagaat
     1861 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1921 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1981 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2041 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2101 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2161 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2221 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2281 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2341 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2401 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2461 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2521 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2581 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2641 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg
     2701 cagggcctgc cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt
     2761 ggggtgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     2821 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt
     2881 gcgttttatt ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct
     2941 tccgtgtttc agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc
     3001 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     3061 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     3121 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     3181 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     3241 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     3301 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     3361 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc
     3421 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     3481 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     3541 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     3601 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     3661 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     3721 ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac cggacaggtc
     3781 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     3841 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     3901 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     3961 atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     4021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     4081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     4141 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     4201 actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     4261 gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg ttctttccgc
     4321 ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4441 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4741 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4801 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4861 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttacgcgtt
     4921 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     4981 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5041 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     5101 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt tatctagatc
     5161 cggtggatcc cgggcccgcg gtaccgtcga ctgcagaatt cgaagcttga gctcgagatc
     5221 tgagtccgga cttgtacact aggcgaagat gcccccaaca gtggaggcga tgacgatgcc
     5281 caggatcaca cagcagatga tgatcatgat tttcttccgg cgcgccttgc tctggtactt
     5341 gacggccttc ttggtgtcag acgatcgctc gtcc
//
