LOCUS       sequence-182522-        5641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of syntaxin-4 C-terminally conjugated to mCitrine..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5641)
  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 5641)
  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: "Elife."; 
            date: "2017-05-19"; volume: "6. pii: e23525. doi"; pages: " 
            10.7554/eLife.23525"
COMMENT     Sequence Label: Stx4-pmCitrine
FEATURES             Location/Qualifiers
     source          1..5641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     regulatory      1582..1591
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1588..1590,1591..1593,1594..2307)
                     /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(1894..1913)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2429..2550
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2466..2485)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2520..2539
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2557..3012)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2694..2715)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2906..2925
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3039..3143
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3145..3502
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3145..3336
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3168..3188)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3353..3488
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3537..4331
                     /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             3537..4331
                     /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(3591..3610)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4201..4220
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4519..4538)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4563..4610
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4939..5527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5428..5447
                     /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 atctcgagct caagcttcga attctgcagt cgacggtacc cgccaccatg
      661 cgcgacagga cccatgagtt gaggcagggg gataacatct cagacgatga agatgaggtt
      721 cgagtcgcgt tggtggtgca ctcaggtgcc gcccggctga gcagcccgga cgacgagttc
      781 ttccagaagg tgcagacaat tcggcagact atggccaaac tggagagtaa agtccgggag
      841 ttggagaaac agcaggtcac cattctggcc acgcctcttc ccgaggagag catgaagcag
      901 ggcctgcaga acctgcgaga ggagatcaaa cagctgggga gagaagtccg ggcacagcta
      961 aaagccatag agccccagaa ggaagaagct gatgagaatt ataattcagt caacacaaga
     1021 atgaagaaaa cccagcatgg ggtcctgtcc cagcagtttg tggagctcat caacaagtgt
     1081 aactcaatgc agtccgagta ccgagagaag aacgtagagc gcattcggcg gcagctgaag
     1141 atcaccaatg ctggaatggt gtctgacgag gaactggagc agatgctgga cagtgggcag
     1201 agtgaggtgt ttgtgtctaa tatactgaag gacacacagg tgacccggca ggccctgaat
     1261 gagatctctg cgcgacacag tgagatccag cagttggagc gcagcatccg tgaactccat
     1321 gagatcttca cttttctagc taccgaggtg gagatgcagg gagagatgat caatcgtatc
     1381 gaaaagaaca ttctgagctc agcagactat gtggaacgtg ggcaagaaca tgtcaagata
     1441 gcgctagaga atcagaagaa ggcgaggaag aaaaaggtca tgattgccat ctgtgtttct
     1501 gtcactgttc tcatcttggc tgtcatcatt ggcatcacca taaccgttgg attggttccg
     1561 cgggcccggg atccacccgt ggccaccatg gtgagcaagg gcgaggagct gttcaccggc
     1621 gtggtgccca tcctggtgga gctggacggc gacgtgaacg gccacaagtt cagcgtgagc
     1681 ggcgagggcg agggcgacgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     1741 ggcaagctgc ccgtgccctg gcccaccctg gtgaccacct tcggctacgg cctgatgtgc
     1801 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagagcgc catgcccgag
     1861 ggctacgtgc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     1921 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     1981 aaggaggacg gcaacatcct gggccacaag ctggagtaca actacaacag ccacaacgtg
     2041 tacatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     2101 atcgaggacg gcagcgtgca gctggccgac cactaccagc agaacacccc catcggcgac
     2161 ggccccgtgc tgctgcccga caaccactac ctgagctacc agagcaagct gagcaaggac
     2221 cccaacgaga agcgcgacca catggtgctg ctggagttcg tgaccgccgc cggcatcacc
     2281 ctgggcatgg acgagctgta caagtaagcg gccgcgactc tagatcataa tcagccatac
     2341 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2401 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2461 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2521 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     2581 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2641 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2701 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2761 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2821 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2881 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2941 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3001 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3061 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3121 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3181 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3241 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3301 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3361 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3421 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3481 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3541 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3601 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3661 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     3721 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3781 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3841 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3901 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3961 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4021 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4081 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4141 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4201 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4261 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4321 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4381 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4441 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4501 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4561 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     4621 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4681 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4741 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     4801 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4861 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4921 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4981 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5041 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5101 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5161 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5221 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5281 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5341 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5401 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5461 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5521 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5581 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     5641 t
//
