LOCUS       Exported                5982 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of syntaxin-3 N-terminally conjugated to FKBP and 
            C-terminally conjugated to mCitrine..
ACCESSION   .
VERSION     .
KEYWORDS    FKBP-Stx3-mCitrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5982)
  AUTHORS   Verboogen DRJ, Gonzalez Mancha N, Ter Beest M, van den Bogaart G
  TITLE     Fluorescence lifetime imaging microscopy reveals rerouting of SNARE 
            trafficking driving dendritic cell activation.
  JOURNAL   Elife. 2017 May 19;6. pii: e23525. doi: 10.7554/eLife.23525.
  PUBMED    28524818
REFERENCE   2  (bases 1 to 5982)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5982
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             624..947
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             1602..1613
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      1923..1932
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1929..2648
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2235..2254)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2770..2891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2807..2826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2861..2880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2898..3353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3035..3056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3247..3266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3380..3484
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3486..3843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3509..3529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3694..3829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3756..3775
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3878..4672
                     /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(3932..3951)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4542..4561
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4860..4879)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4904..4951
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5280..5868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5769..5788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc accatgggcg tgcaggtgga gaccatcagc cccggcgacg
      661 gccgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggc atgctggagg
      721 acggcaagaa gttcgacagc agccgcgacc gcaacaagcc cttcaagttc atgctgggca
      781 agcaggaggt gatccgcggc tgggaggagg gcgtggccca gatgagcgtg ggccagcgcg
      841 ccaagctgac catcagcccc gactacgcct acggcgccac cggccacccc ggcatcatcc
      901 ccccccacgc caccctggtg ttcgacgtgg agctgctgaa gctggagtct ggtggcggtg
      961 gctcgggcgg aggtgggtcg ggtggcggcg gaagaattct gcagtcgacg gtacccgcca
     1021 ccatgaagga ccgactggag cagctgaagg ccaagcagct gacgcaggat gatgacacgg
     1081 acgaggttga gattgctatt gataatacag cgttcatgga tgagttcttt tctgagattg
     1141 aggaaacgag gctcaacatc gacaagatct cagagcatgt ggaggaagct aagaaactct
     1201 acagtatcat tctctctgca ccgattccag agccaaaaac caaagacgac cttgaacagc
     1261 tcacaactga gatcaagaaa agggccaaca acgtccggaa caaactgaag agcatggaga
     1321 agcatattga ggaagacgag gtccggtcat cagcagacct tcggatacga aagtcccagc
     1381 actccgtcct ctcccggaag tttgtggagg tgatgacaaa gtacaatgag gcacaggtgg
     1441 acttccgtga acgcagcaaa gggcgcatcc aacggcagct tgaaattact ggcaagaaaa
     1501 caacagatga ggagctggaa gaaatgttgg agagtggcaa cccagccatc ttcacttctg
     1561 ggatcattga ctcccagatt tccaagcaag ccctcagcga gattgagggt cggcacaagg
     1621 acatcgtgag gctggagagc agcatcaagg agctccatga catgtttatg gacatcgcca
     1681 tgctggtgga gaatcagggt gagatgttag ataacataga gttgaatgtc atgcacacgg
     1741 tggaccatgt ggagaaggca cgggatgaaa ctaaaagagc catgaagtat cagggtcagg
     1801 ctcgaaagaa attgataatt atcattgtga tagtagttgt gttgctgggc attttagcgt
     1861 tgattattgg attgtccgtt gggctgaagt cgacggtacc gcgggcccgg gatccacccg
     1921 tggccaccat ggtgagcaag ggcgaggagc tgttcaccgg cgtggtgccc atcctggtgg
     1981 agctggacgg cgacgtgaac ggccacaagt tcagcgtgag cggcgagggc gagggcgacg
     2041 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2101 ggcccaccct ggtgaccacc ttcggctacg gcctgatgtg cttcgcccgc taccccgacc
     2161 acatgaagca gcacgacttc ttcaagagcg ccatgcccga gggctacgtg caggagcgca
     2221 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2281 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2341 tgggccacaa gctggagtac aactacaaca gccacaacgt gtacatcatg gccgacaagc
     2401 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2461 agctggccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2521 acaaccacta cctgagctac cagagcaagc tgagcaagga ccccaacgag aagcgcgacc
     2581 acatggtgct gctggagttc gtgaccgccg ccggcatcac cctgggcatg gacgagctgt
     2641 acaagtaagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     2701 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2761 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2821 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2881 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2941 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3001 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3061 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3121 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3181 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3241 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3301 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3361 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3421 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3481 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3661 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3721 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3781 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3841 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3901 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3961 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4021 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4081 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4141 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4201 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4261 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4321 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4381 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4441 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4501 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4561 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4621 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4681 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4741 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4801 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4861 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4921 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4981 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5041 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5101 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5161 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5221 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5281 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5341 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5401 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5461 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5521 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5581 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5641 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5701 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5761 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5821 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5881 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5941 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
