LOCUS       sequence-239356-        6939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synchronize trafficking of WNT3A from the ER (RUSH System).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6939)
  AUTHORS   Moti N, Yu J, Boncompain G, Perez F, Virshup DM
  TITLE     Wnt traffic from endoplasmic reticulum to filopodia.
  JOURNAL   PLoS One. 2019 Feb 22;14(2):e0212711. doi: 
            10.1371/journal.pone.0212711. eCollection 2019.
  PUBMED    30794657
REFERENCE   2  (bases 1 to 6939)
  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: "2019-02-22"; volume: "14(2):e0212711. doi"; pages: " 
            10.1371/journal.pone.0212711. eCollection 2019"
COMMENT     Sequence Label: Str-KDEL_SBP-EGFP-WNT3A
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1646..1875
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1858..1877
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2039..2501
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2095..2112)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2322..2341
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     sig_peptide     2528..2587
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             2588..2698
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             join(2708..2710,2711..3421)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTLAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2750..2771)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3011..3030)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3358..3379
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4559..4680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4596..4615)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4782..4798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4782..4798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4795..4817)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4806..4822
                     /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(4830..4836,4837..4854,4855..4860))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4875..4896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5013..5030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5184..5772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5264..5283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5943..6734,6735..6803))
                     /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     6566..6585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6804..6908)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtatggatg
      961 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
     1021 tcagccatag tcacagtgag aaggcgacag gaaccagctc gggcgccggt ttaaacgacc
     1081 ctagcaaaga ctcaaaagct caggtgtccg ctgccgaggc tggcattact ggaacatggt
     1141 acaatcagct cgggagcacc tttattgtga ctgctggagc cgatggagcc ctcaccggaa
     1201 catacgaatc tgctgtggga aacgccgaat cacggtacgt cctcactggc cgatacgata
     1261 gtgcccctgc caccgacgga tctgggactg ccctgggatg gactgtcgct tggaaaaaca
     1321 actaccggaa tgctcattct gccacaacat ggagtggaca gtacgtggga ggcgctgagg
     1381 ctagaatcaa tacacagtgg ctgctcacat ctggcacaac cgaggcaaat gcttggaaat
     1441 ccaccctggt gggacatgac acattcacca aagtgaaacc ctccgccgct tcaatcgatg
     1501 ccgccaaaaa agccggagtc aacaacggca atcctctgga tgccgtccag cagctcggac
     1561 cgaaagatga actgtgagcg gccgcataga taactgatcc agtgtgctgg aattaattcg
     1621 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
     1681 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
     1741 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
     1801 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
     1861 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
     1921 cggccaattc cgcccctctc ccccccccta acgttactgg ccgaagccgc ttggaataag
     1981 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     2041 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     2101 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     2161 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     2221 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     2281 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     2341 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     2401 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     2461 accacgggga cgtggttttc ctttgaaaaa cacgatgata agcttgccac aacccgggag
     2521 gcgcgccatg tacaggatgc aactcctgtc ttgcattgca ctaagtcttg cacttgtcac
     2581 gaattccgac gagaagacca ctggttggcg aggtggacac gttgttgaag gactggctgg
     2641 ggaacttgaa caacttcgtg cacgactgga gcatcaccca caaggtcaac gtgaaccacc
     2701 tgcaggtgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2761 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2821 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2881 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2941 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3001 cttcttcaag gacgacggca actacaagac cctcgccgag gtgaagttcg agggcgacac
     3061 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3121 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     3181 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     3241 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3301 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3361 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3421 gggccggcca tacccgatct ggtggtcgct ggctgttggg ccacagtatt cctccctggg
     3481 ctcgcagccc atcctgtgtg ccagcatccc gggcctggtc cccaagcagc tccgcttctg
     3541 caggaactac gtggagatca tgcccagcgt ggccgagggc atcaagattg gcatccagga
     3601 gtgccagcac cagttccgcg gccgccggtg gaactgcacc accgtccacg acagcctggc
     3661 catcttcggg cccgtgctgg acaaagctac cagggagtcg gcctttgtcc acgccattgc
     3721 ctcagccggt gtggcctttg cagtgacacg ctcatgtgca gaaggcacgg ccgccatctg
     3781 tggctgcagc agccgccacc agggctcacc aggcaagggc tggaagtggg gtggctgtag
     3841 cgaggacatc gagtttggtg ggatggtgtc tcgggagttc gccgacgccc gggagaaccg
     3901 gccagatgcc cgctcagcca tgaaccgcca caacaacgag gctgggcgcc aggccatcgc
     3961 cagccacatg cacctcaagt gcaagtgcca cgggctgtcg ggcagctgcg aggtgaagac
     4021 atgctggtgg tcgcaacccg acttccgcgc catcggtgac ttcctcaagg acaagtacga
     4081 cagcgcctcg gagatggtgg tggagaagca ccgggagtcc cgcggctggg tggagaccct
     4141 gcggccgcgc tacacctact tcaaggtgcc cacggagcgt gacctggtct actacgaggc
     4201 ctcgcccaac ttctgcgagc ccaaccctga gacgggctcc ttcggcacgc gcgaccgcac
     4261 ctgcaacgtc agctcgcacg gcatcgacgg ctgcgacctg ctgtgctgcg gccgcggcca
     4321 caacgcgcga gcggagcggc gccgggagaa gtgccgctgc gtgttccact ggtgctgcta
     4381 cgtcagctgc caggagtgca cgcgcgtcta cgacgtgcac acctgcaagt agttaattaa
     4441 ctctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4501 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4561 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4621 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4681 acgcgtcgag tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4741 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4801 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4861 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4921 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4981 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5041 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5101 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5161 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5221 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5281 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5341 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5401 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5461 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5521 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5581 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5641 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5701 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6901 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
