LOCUS       sequence-339113-        8917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For light regulated release of GluA1 from the endoplasmic reticulum.
            Driven by synapsin promoter. Intracellular trafficking can be better
            visualized with mNeon..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8917)
  AUTHORS   Bourke AM, Schwartz SL, Bowen AB, Kleinjan MS, Winborn CS, Kareemo 
            DJ, Gutnick A, Schwarz TL, Kennedy MJ
  TITLE     zapERtrap: A light-regulated ER release system reveals unexpected 
            neuronal trafficking pathways.
  JOURNAL   J Cell Biol. 2021 Sep 6;220(9). pii: 212461. doi: 
            10.1083/jcb.202103186. Epub 2021 Jul 9.
  PUBMED    34241635
REFERENCE   2  (bases 1 to 8917)
  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: "J Cell 
            Biol."; date: "2021-09-6"; volume: "220(9). pii: 212461. doi"; 
            pages: " 10.1083/jcb.202103186. Epub 2021 Jul 9"
COMMENT     Sequence Label: DHFR-mNeon-GluA1
FEATURES             Location/Qualifiers
     source          1..8917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          636..1652
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1576..1598
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1660..1679
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1712..1721
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1718..1795
                     /codon_start=1
                     /product="human growth hormone signal sequence"
                     /label=hGH signal sequence
                     /translation="MATGSRTSLLLAFGLLCLPWLQEGSA"
     CDS             1802..2275
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             2309..3016
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      2564..2573
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3041..3067
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5905..5924)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5970..6025
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6024..6043)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6416..6432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6416..6432)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6429..6451)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6440..6456
                     /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(6464..6470,6471..6488,6489..6494))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6509..6530
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6589..6784
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6635..6770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6790..6924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6827..6846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6881..6900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6992..7009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7163..7751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7243..7262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7922..8713,8714..8782))
                     /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     8545..8564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8783..8887)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactaga gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat
      421 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc
      481 gatgggggcg gggggggggg gggggggagg ggcggggcgg ggcgaggcgg agaggtgcgg
      541 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
      601 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
      661 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      721 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
      781 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
      841 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
      901 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
      961 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     1021 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     1081 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     1141 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     1201 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     1261 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     1321 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     1381 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     1441 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     1501 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     1561 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     1621 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     1681 tgtctcatca ttttggcaaa gaattgctcg agccaccatg gctacaggct cccggacgtc
     1741 cctgctcctg gcttttggcc tgctctgcct gccctggctt caagagggca gtgccgagct
     1801 catcagtctg attgcggcgt tagcggtaga tcgcgttatc ggcatggaaa acgccatgcc
     1861 gtggaacctg cctgccgatc tcgcctggtt taaacgcaac accttaaata aacccgtgat
     1921 tatgggccgc catacctggg aatcaatcgg tcgtccgttg ccaggacgca aaaatattat
     1981 cctcagcagt caaccgggta cggacgatcg cgtaacgtgg gtgaagtcgg tggatgaagc
     2041 catcgcggcg tgtggtgacg taccagaaat catggtgatt ggcggcggtc gcgtttatga
     2101 acagttcttg ccaaaagcgc aaaaactgta tctgacgcat atcgacgcag aagtggaagg
     2161 cgacacccat ttcccggatt acgagccgga tgactgggaa tcggtattca gcgaattcca
     2221 cgatgctgat gcacagaact ctcacagcta ttgctttgag attctggagc ggcgggtcga
     2281 cagtaaagga gaagaacttt tcactggaat ggtgagcaag ggcgaggagg ataacatggc
     2341 ctctctccca gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga
     2401 catggtgggt cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctgaagtc
     2461 caccaagggt gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt
     2521 ccatcagtac ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg
     2581 ctccggctac caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa
     2641 ctaccgctac acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg
     2701 tttccctgct gacggtcctg tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc
     2761 gaagaagact taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac
     2821 tggaaatggc aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat
     2881 ggcggctaac tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca
     2941 ctccaagacc gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat
     3001 ggacgagctg tacaagagtg ctggtagtgc tggtaagctt tacccatacg atgttccaga
     3061 ttacgctacg cgtggaggtg gagggggcgg aggtggcggg ggttcgcgct tccccaacaa
     3121 tatccagata ggggggttat ttccaaacca acaatcacag gaacatgcgg cttttaggtt
     3181 tgctttgtca caactcacgg agccccccaa gctgcttccc cagatcgata ttgtgaacat
     3241 cagcgacagc tttgagatga cttaccgttt ctgttcccag ttctccaaag gagtctatgc
     3301 catctttgga ttttatgaac gaaggactgt caacatgctg acctccttct gtggggccct
     3361 ccatgtgtgc ttcattactc caagttttcc tgttgacaca tccaatcaat ttgtccttca
     3421 gctacgcccg gaactacagg aagctctcat tagcattatc gaccattaca agtggcaaac
     3481 ctttgtctac atttatgatg ctgaccgggg cctgtcagtc ctgcagagag tcttggatac
     3541 agccgcagag aagaactggc aggtaacggc tgtcaacatt ctgacaacca ccgaggaagg
     3601 ataccggatg ctctttcagg acctggagaa gaaaaaggag aggctggtgg tggttgactg
     3661 tgaatcagaa cgcctcaacg ccatcctggg ccagatcgtg aagctagaaa agaatggcat
     3721 cgggtaccac tacatcctcg ccaatctggg cttcatggac attgacttaa ataagttcaa
     3781 ggagagcgga gccaatgtga caggtttcca gctggtgaac tacacagaca cgatcccagc
     3841 cagaatcatg cagcaatgga ggacaagtga ctcccgagac cataccaggg tggactggaa
     3901 gaggccaaag tacacttctg ctctcaccta tgatggtgtc aaggtgatgg ctgaggcctt
     3961 ccaaagcctg cggaggcaga ggattgacat atcccgccgg gggaatgctg gggactgtct
     4021 ggctaaccca gctgtgccct ggggtcaagg gatcgacatc cagagagccc tgcagcaggt
     4081 gcgcttcgaa ggtttgacag gaaatgtgca gttcaacgag aaagggcgcc ggaccaatta
     4141 caccctccac gtgatcgaaa tgaaacatga tggaatccga aagattggtt actggaatga
     4201 agacgataaa tttgtccccg cagccaccga cgctcaggct ggaggggaca actcaagcgt
     4261 ccagaatagg acctacatcg tcactactat cctcgaagat ccttacgtga tgcttaaaaa
     4321 gaatgccaac cagtttgagg gcaatgaccg ctatgagggc tactgtgtgg agctggctgc
     4381 agagatcgcc aagcacgtgg gctactccta ccgacttgag attgtcagcg acggcaaata
     4441 tggagcccgg gatcccgaca caaaggcttg gaatggcatg gtgggagaac tggtctatgg
     4501 aagagcagac gtggctgtgg ctcccttgac cataaccttg gtccgggagg aagtcatcga
     4561 cttctccaag ccattcatga gtttgggaat ctccattatg attaagaagc cacagaagtc
     4621 caagccaggt gtcttctcct ttcttgaccc tttggcctat gagatctgga tgtgtatagt
     4681 gtttgcctac attggagtga gcgtcgtcct cttcctggtc agccgtttca gcccctacga
     4741 atggcacagc gaagagtttg aagagggacg agaccagaca accagtgacc agtcaaatga
     4801 gtttggcata ttcaacagcc tgtggttctc cctgggggcc ttcatgcagc aaggatgtga
     4861 catttccccc aggtccctgt ccggacgcat cgtcggcggc gtctggtggt tcttcacttt
     4921 gatcatcatc tcctcgtaca cagccaacct ggctgccttc ctgactgtgg agaggatggt
     4981 gtctcccatt gagagtgcag aggacctggc aaagcagacg gaaattgctt atgggacatt
     5041 ggaagcaggc tccactaagg agttcttcag gagatctaaa atcgctgtgt ttgagaagat
     5101 gtggacatac atgaagtctg cagaaccatc cgtgtttgtt cggaccacag aggaaggcat
     5161 gatcagagtg agaaaatcta aaggcaaata cgcctacctc ctggagtcca ccatgaatga
     5221 gtatattgag caacgaaagc cctgtgacac catgaaagtg ggaggtaact tggattccaa
     5281 aggctatggc attgcgacac ccaaggggtc cgccctgaga aatccagtaa acctggcagt
     5341 gttaaaactg aacgagcagg ggcttttgga caaattgaaa aacaaatggt ggtacgacaa
     5401 gggcgagtgc ggcagcgggg gaggtgactc caaggacaag accagcgctt tgagcctcag
     5461 caatgtggca ggcgtgttct acatcctgat tggagggctg ggactggcca tgctggttgc
     5521 cttaatcgag ttctgctaca aatcccgtag cgagtcgaag cggatgaagg gtttctgttt
     5581 gatcccacag caatccatca atgaagccat acggacatcg accctccccc ggaacagtgg
     5641 ggcaggagcc agcggaggag gcggcagtgg agagaatggc cgggtggtca gccaggactt
     5701 ccccaagtcc atgcaatcca ttccctgcat gagtcacagt tcagggatgc ccttgggagc
     5761 cacaggattg taagaattct gcagtcgacg gtaccgcggg cccgtacaag taaagcggcc
     5821 gcaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     5881 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     5941 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     6001 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     6061 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     6121 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     6181 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     6241 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     6301 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     6361 gcaccgtcga ccagctggtc gacggtgcac cgtcgaccag cttggcgtaa tcatggtcat
     6421 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6481 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6541 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta
     6601 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6661 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6721 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6781 caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6841 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6901 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
     6961 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7021 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7081 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7141 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7201 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7261 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7321 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7381 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7441 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7501 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7561 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7621 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7681 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7741 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7801 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7861 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7921 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7981 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8041 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8101 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8161 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     8221 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8281 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8341 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8401 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8461 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8521 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8581 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8641 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8701 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8761 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8821 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8881 ttccgcgcac atttccccga aaagtgccac ctgggtc
//
