LOCUS       sequence-369209-        6543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Stim1 with C-terminal domain deletion fused to YFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6543)
  AUTHORS   Zamponi E, Meehl JB, Voeltz GK
  TITLE     The ER ladder is a unique morphological feature of developing 
            mammalian axons.
  JOURNAL   Dev Cell. 2022 Jun 6;57(11):1369-1382.e6. doi: 
            10.1016/j.devcel.2022.05.002. Epub 2022 May 23.
  PUBMED    35609616
REFERENCE   2  (bases 1 to 6543)
  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"; doi: 
            "10.1016/j.devcel.2022.05.002"; journalName: "Dev Cell"; date: 
            "2022-06-6- 6"; volume: "57"; issue: "11"; pages: "1369-1382.e6"
COMMENT     Sequence Label: YFP-Stim1-Delta-CTD
FEATURES             Location/Qualifiers
     source          1..6543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(624..626,627..629,630..1340)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(669..690)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(930..949)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1277..1298
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    1350..1374
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3271..3392
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3308..3327)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3362..3381
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3399..3854)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3536..3557)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3748..3767
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3881..3985
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3987..4344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3987..4178
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4010..4030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4195..4330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4379..5173
                     /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             4379..5173
                     /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(4433..4452)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5043..5062
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5361..5380)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5405..5452
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5781..6369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6270..6289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      541 tggatgtatg cgtccgtctt gccctgtggc tcctctgggg actcctcctg caccagggcc
      601 agagcctcgc accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      661 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      721 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      781 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg
      841 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      901 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      961 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1021 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1081 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1141 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1201 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     1261 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1321 gcatggacga gctgtacaag tccggatcaa caagtttgta caaaaaagca ggctccctca
     1381 gccatagtca cagtgagaag gcgacaggaa ccagctcggg ggccaactct gaggagtcca
     1441 ctgcagcaga gttttgccga attgacaagc ccctgtgtca cagtgaggat gagaaactca
     1501 gcttcgaggc agtccgtaac atccacaaac tgatggacga tgatgccaat ggtgatgtgg
     1561 atgtggaaga aagtgatgag ttcctgaggg aagacctcaa ttaccatgac ccaacagtga
     1621 aacacagcac cttccatggt gaggataagc tcatcagcgt ggaggacctg tggaaggcat
     1681 ggaagtcatc agaagtatac aattggaccg tggatgaggt ggtacagtgg ctgatcacat
     1741 atgtggagct gcctcagtat gaggagacct tccggaagct gcagctcagt ggccatgcca
     1801 tgccaaggct ggctgtcacc aacaccacca tgacagggac tgtgctgaag atgacagacc
     1861 ggagtcatcg gcagaagctg cagctgaagg ctctggatac agtgctcttt gggcctcctc
     1921 tcttgactcg ccataatcac ctcaaggact tcatgctggt ggtgtctatc gttattggtg
     1981 tgggcggctg ctggtttgcc tatatccaga accgttactc caaggagcac atgaagaaga
     2041 tgatgaagga cttggagggg ttacaccgag ctgagcagag tctgcatgac cttcaggaaa
     2101 ggctgcacaa ggcccaggag gagcaccgca cagtggaggt ggagaaggtc catctggaaa
     2161 agaagctgcg cgatgagatc aaccttgcta agcaggaagc ccagcggctg aaggagctgc
     2221 gggagggtac tgagaatgag cggagccgcc aaaaatatgc tgaggaggag ttggagcagg
     2281 ttcgggaggc cttgaggaaa gcagagaagg agctagaatc tcacagctca tggtatgctc
     2341 cagaggccct tcagaagtgg ctgcagctga cacatgaggt ggaggtgcaa tattacaaca
     2401 tcaagaagca aaatgctgag aagcagctgc tggtggccaa ggagggggct gagaagataa
     2461 aaaagaagag aaacacactc tttggcacct tccacgtggc ccacagctct tccctggatg
     2521 atgtagatca taaaattcta acagctaagc aagcactgag cgaggcgaca gcagcattgc
     2581 gggagcgcct gcaccgctgg caacagatcg agatcctctg tggcttccag attgtcaaca
     2641 accctggcat ccactcactg gtggctgccc tcaacataga ccccagctgg atgggcagta
     2701 cacgccccaa ccctgctcac ttcatcatga ctgacgacgt ggatgacatg gatgaggaga
     2761 ttgtgtctcc cttgtccatg cagtccccta gcctgcagag cagtgttcgg cagcgcctga
     2821 cggagccaca gcatggcctg ggatctcaga gggatttgac ccattccgat tcggagtcct
     2881 ccctccacat gagtgaccgc cagcgtgtgg cccccaaacc tcctcagatg agccgtgctg
     2941 cagacgaggc tctcaatgcc atgacttcca atggcagcca ccggctgatc gagggggtcc
     3001 acccagggtc tctggtggag aaactgcctg acagccctgc cctggccaag aaggcattac
     3061 tggcgctgaa ccatgggctg gacaaggccc acagcctgat ggagctgtag gaattctgca
     3121 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     3181 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3241 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3301 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3361 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3421 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3481 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3541 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3601 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3661 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3721 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3781 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3841 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3901 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3961 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4021 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4081 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4141 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4201 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4261 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4321 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4381 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4441 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4501 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4561 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4621 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4681 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4741 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4801 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4861 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4921 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4981 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     5041 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5101 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5161 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     5221 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5281 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5341 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5401 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5461 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5521 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5581 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5641 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5701 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5761 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5821 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5881 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5941 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6001 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6061 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6121 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6181 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6241 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6301 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6361 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6421 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6481 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6541 ccg
//
