LOCUS       sequence-173842-        6005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Stx3 with triple mutation in the apical targeting 
            signal (F31A D33A E34A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6005)
  AUTHORS   Soo Hoo L, Banna CD, Radeke CM, Sharma N, Albertolle ME, Low SH, 
            Weimbs T, Vandenberg CA
  TITLE     The SNARE Protein Syntaxin 3 Confers Specificity for Polarized 
            Axonal Trafficking in Neurons.
  JOURNAL   PLoS One. 2016 Sep 23;11(9):e0163671. doi: 
            10.1371/journal.pone.0163671. eCollection 2016.
  PUBMED    27662481
REFERENCE   2  (bases 1 to 6005)
  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: "PLoS One.";
            date: "2016-09-23"; volume: "11(9):e0163671. doi"; pages: " 
            10.1371/journal.pone.0163671. eCollection 2016"
COMMENT     Sequence Label: Stx3 triple mutant (F31A D33A E34A)
FEATURES             Location/Qualifiers
     source          1..6005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             580..591
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             892..921
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             937..954
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(977..994)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    983..1207
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1253..1681
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1340..1359)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1550..1571
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1690..1710)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1695..2024
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1875..2010
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1937..1956
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2072..2119
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2138..2512
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2642..2763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2679..2698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2733..2752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2812..2828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2812..2828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2825..2847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2836..2852
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2860..2866,2867..2884,2885..2890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2905..2926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3043..3060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3214..3802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3294..3313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3973..4764,4765..4833))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4596..4615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4834..4938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5013..5032)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5204..5583
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5584..5787
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5738..5758
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    5789..5807
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5810..5828
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     5838..5862
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 atgaaggacc gtctggagca gctgaaggcc aagcagctga cacaggatga tgatactgat
       61 gcggttgaga ttgctatcga caacacggct gctatggccg cgttcttttc tgagattgag
      121 gaaactcggc ttaacattga caagatctca gaacatgtag aggaggctaa gaaactctac
      181 agtatcattc tctctgcacc gattccagag ccaaaaacca aggatgacct agagcagctc
      241 acgactgaga ttaagaaaag ggccaacaac gtccggaaca aactgaagag catggagaag
      301 catattgaag aagatgaggt caggtcatcg gcagaccttc ggattcggaa atcccagcac
      361 tctgtccttt ctcggaagtt tgtggaggtg atgaccaaat acaatgaagc tcaagtggac
      421 ttccgagaac gcagcaaagg gcgaatccag cggcagctcg aaattactgg caaaaagaca
      481 accgatgagg agctggagga gatgttggag agtggcaacc cggccatctt cacttctggg
      541 atcattgact cacagatttc caagcaagcc ctcagtgaga ttgagggacg acacaaggac
      601 attgtgaggc tggagagcag catcaaggag cttcacgaca tgtttatgga catcgccatg
      661 ctggtggaga atcagggtga gatgttagat aacatagagt tgaatgtcat gcacacagtg
      721 gaccacgtgg agaaggcacg agatgaaacg aaaaaagctg tgaaatacca gagtcaggcc
      781 cggaagaaat tgataattat cattgtgcta gtagttgtgt tgctgggcat tttagcattg
      841 attattggac tttccgttgg gctgaatgcg gccgctcgag gtcacccatt cgaacaaaaa
      901 ctcatctcag aagaggatct gaatatgcat accggtcatc atcaccatca ccattgagtt
      961 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1021 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1081 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1141 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1201 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     1261 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1321 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1381 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1441 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1501 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1561 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1621 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1681 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     1741 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     1801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     1861 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     1921 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     1981 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     2041 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     2101 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     2161 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     2221 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     2281 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     2341 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     2401 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     2461 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     2521 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     2581 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     2641 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2701 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2761 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     2821 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2881 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2941 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3001 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3061 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3121 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3841 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3901 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3961 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4141 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4261 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4501 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4561 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4801 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4861 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     4981 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5041 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     5101 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     5161 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     5221 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5281 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5341 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5401 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5461 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5521 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5581 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5641 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5701 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     5761 cggtgggagg tctatataag cagagctctc cctatcagtg atagagatct ccctatcagt
     5821 gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
     5881 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac tctagcgttt
     5941 aaacttaagc ttggtaccga gctcggatcc actagtccag tgtggtggaa ttcctgggct
     6001 tcagg
//
