LOCUS       Exported                6006 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of syntaxin-4 N-terminally conjugated to FKBP and 
            C-terminally conjugated to mCitrine..
ACCESSION   .
VERSION     .
KEYWORDS    FKBP-Stx4-mCitrine
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6006)
  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 6006)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6006
                     /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"
     regulatory      1947..1956
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1953..2672
                     /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(2259..2278)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2794..2915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2831..2850)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2885..2904
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2922..3377)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3059..3080)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3271..3290
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3404..3508
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3510..3867
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3533..3553)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3718..3853
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3780..3799
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3902..4696
                     /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(3956..3975)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4566..4585
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4884..4903)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4928..4975
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5304..5892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5793..5812
                     /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 ccatgcgcga caggacccat gagttgaggc agggggataa catctcagac gatgaagatg
     1081 aggttcgagt cgcgttggtg gtgcactcag gtgccgcccg gctgagcagc ccggacgacg
     1141 agttcttcca gaaggtgcag acaattcggc agactatggc caaactggag agtaaagtcc
     1201 gggagttgga gaaacagcag gtcaccattc tggccacgcc tcttcccgag gagagcatga
     1261 agcagggcct gcagaacctg cgagaggaga tcaaacagct ggggagagaa gtccgggcac
     1321 agctaaaagc catagagccc cagaaggaag aagctgatga gaattataat tcagtcaaca
     1381 caagaatgaa gaaaacccag catggggtcc tgtcccagca gtttgtggag ctcatcaaca
     1441 agtgtaactc aatgcagtcc gagtaccgag agaagaacgt agagcgcatt cggcggcagc
     1501 tgaagatcac caatgctgga atggtgtctg acgaggaact ggagcagatg ctggacagtg
     1561 ggcagagtga ggtgtttgtg tctaatatac tgaaggacac acaggtgacc cggcaggccc
     1621 tgaatgagat ctctgcgcga cacagtgaga tccagcagtt ggagcgcagc atccgtgaac
     1681 tccatgagat cttcactttt ctagctaccg aggtggagat gcagggagag atgatcaatc
     1741 gtatcgaaaa gaacattctg agctcagcag actatgtgga acgtgggcaa gaacatgtca
     1801 agatagcgct agagaatcag aagaaggcga ggaagaaaaa ggtcatgatt gccatctgtg
     1861 tttctgtcac tgttctcatc ttggctgtca tcattggcat caccataacc gttggattgg
     1921 ttccgcgggc ccgggatcca cccgtggcca ccatggtgag caagggcgag gagctgttca
     1981 ccggcgtggt gcccatcctg gtggagctgg acggcgacgt gaacggccac aagttcagcg
     2041 tgagcggcga gggcgagggc gacgccacct acggcaagct gaccctgaag ttcatctgca
     2101 ccaccggcaa gctgcccgtg ccctggccca ccctggtgac caccttcggc tacggcctga
     2161 tgtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag agcgccatgc
     2221 ccgagggcta cgtgcaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2281 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2341 acttcaagga ggacggcaac atcctgggcc acaagctgga gtacaactac aacagccaca
     2401 acgtgtacat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     2461 acaacatcga ggacggcagc gtgcagctgg ccgaccacta ccagcagaac acccccatcg
     2521 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagagc aagctgagca
     2581 aggaccccaa cgagaagcgc gaccacatgg tgctgctgga gttcgtgacc gccgccggca
     2641 tcaccctggg catggacgag ctgtacaagt aagcggccgc gactctagat cataatcagc
     2701 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2761 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2821 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2881 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     2941 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3001 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3061 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3121 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3181 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3241 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3301 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3361 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3421 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3481 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3541 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3601 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3661 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3721 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3781 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3841 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3901 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3961 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4021 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4081 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4141 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4201 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4261 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4321 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4381 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4441 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4501 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4561 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4621 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4681 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4741 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4801 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4861 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4921 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4981 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5041 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5101 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5161 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5221 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5281 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5341 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5401 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5461 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5521 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5581 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5641 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5701 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5761 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5821 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5881 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5941 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6001 atgcat
//
