LOCUS       sequence-41533-e        5135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5135)
  AUTHORS   Barg S, Knowles MK, Chen X, Midorikawa M, Almers W
  TITLE     Single secretory granules of live cells recruit syntaxin-1 and 
            synaptosomal associated protein 25 (SNAP-25) in large copy numbers.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Nov 30;107(48):20804-9. Epub 2010 Nov
            12.
  PUBMED    21076040
REFERENCE   2  (bases 1 to 5135)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2010-11-30"; volume: "107(48)"; pages: 
            "20804-9. Epub 2010 Nov 12"
COMMENT     Sequence Label: peGFP(N1)-deltaCMV-rSyntaxin1a-meGFP
FEATURES             Location/Qualifiers
     source          1..5135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1075..1084
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1081..1083,1084..1086,1087..1800)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1126..1147)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1387..1406)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1734..1755
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1923..2044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1960..1979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2014..2033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2051..2506)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2188..2209)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2400..2419
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2533..2637
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2639..2996
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2639..2830
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2662..2682)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2847..2982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3031..3825
                     /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             3031..3825
                     /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(3085..3104)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3695..3714
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4013..4032)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4057..4104
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4433..5021
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4922..4941
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tgggaggtct atataagcag agctggttta
      121 gtgaaccgtc agatccgcta gcgctaccgg actcagatct cgagctcaag cttagccacc
      181 atgaaggacc gaacccagga gctccgcacg gccaaggaca gcgatgacga cgatgatgtc
      241 actgtcactg tggaccgaga ccgcttcatg gatgagttct ttgaacaggt ggaagagatc
      301 cggggcttta ttgacaagat tgctgagaac gtggaggagg tgaagaggaa acacagcgcc
      361 atcctggcct ccccgaaccc cgatgagaag accaaggagg aactggagga gctcatgtcg
      421 gacattaaga agacagcgaa caaagttcgc tccaagctaa agagcatcga gcagagcatc
      481 gagcaggagg aaggtctgaa ccgctcgtcg gcggacctga ggatccggaa gacgcagcat
      541 tccacgctgt cccgaaagtt tgtggaggtc atgtccgagt acaacgccac tcagtcagac
      601 taccgagaac gctgcaaagg gcgcatccag aggcagctgg agatcactgg ccggaccacg
      661 accagtgagg agttggaaga catgctggag agtgggaatc ccgccatctt tgcctctggg
      721 atcatcatgg actccagcat ctcgaagcag gccctcagtg agatcgagac caggcacagt
      781 gagatcatca agttggagaa cagcatccgg gagctacacg atatgttcat ggacatggcc
      841 atgctggtgg agagccaggg ggagatgatt gacaggatcg agtacaatgt ggaacacgct
      901 gtggactacg tggagagggc cgtgtctgac accaagaagg ccgtcaagta ccagagcaag
      961 gcacgcagga agaagatcat gatcatcatt tgctgtgtga ttctgggcat catcatcgcc
     1021 tccaccatcg ggggcatctt tggaggggta ccgcgggccc gggatccacc ggtcgccacc
     1081 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1141 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1201 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1261 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1321 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1381 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1441 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1501 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1561 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1621 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1681 tacctgagca cccagtccaa gcttagcaaa gaccccaacg agaagcgcga tcacatggtc
     1741 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1801 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1861 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1921 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1981 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2041 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2101 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2161 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2221 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2281 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2341 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2401 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2461 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     2521 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2581 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     2641 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2701 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2761 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2821 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2881 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2941 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     3001 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3061 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3121 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3181 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3241 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3301 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3361 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3421 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3481 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3541 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3601 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3661 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3721 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3781 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3841 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3901 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3961 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     4021 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4081 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4141 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4201 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4261 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4321 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4381 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4441 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4501 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4561 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4621 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4681 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4741 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4801 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4861 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4921 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4981 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5041 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5101 ctgattctgt ggataaccgt attaccgcca tgcat
//
