LOCUS       sequence-335646-        8877 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 8877)
  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 8877)
  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..8877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(122..141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(801..1592,1593..1661))
                     /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     1424..1443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1662..1766)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1797..2175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2177..2454
                     /label=chicken beta-actin promoter
     intron          2455..3467
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3391..3413
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3475..3494
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     sig_peptide     3528..3605
                     /codon_start=1
                     /product="human growth hormone signal sequence"
                     /label=hGH signal sequence
                     /translation="MATGSRTSLLLAFGLLCLPWLQEGSA"
     CDS             3612..4085
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             4119..4826
                     /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      4374..4383
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4836..4862
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(7661..7680)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7726..7781
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7780..7799)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8172..8188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8172..8188)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8185..8207)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8196..8212
                     /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(8220..8226,8227..8244,8245..8250))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8265..8286
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8345..8540
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8391..8526
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    8546..8680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8583..8602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8637..8656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8748..8765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
       61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      481 agracagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      601 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      661 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      721 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      781 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      841 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      901 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      961 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1021 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1081 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1141 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1201 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1261 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1321 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1381 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1441 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1501 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1561 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1621 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1681 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1741 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     1801 ttgattattg acagttatta atagtaatca attacggggt cattagttca tagcccatat
     1861 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     1921 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     1981 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2041 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2101 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     2161 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
     2221 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
     2281 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
     2341 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     2401 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     2461 gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc gccccggctc
     2521 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     2581 aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga aagccttaaa
     2641 gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt
     2701 gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     2761 ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg ggcggtgccc
     2821 cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg
     2881 gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac ccccctcccc
     2941 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     3001 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
     3061 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
     3121 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
     3181 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
     3241 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
     3301 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     3361 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     3421 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     3481 tgctggttat tgtgctgtct catcattttg gcaaagaatt gctcgagatg gctacaggct
     3541 cccggacgtc cctgctcctg gcttttggcc tgctctgcct gccctggctt caagagggca
     3601 gtgccgagct catcagtctg attgcggcgt tagcggtaga tcgcgttatc ggcatggaaa
     3661 acgccatgcc gtggaacctg cctgccgatc tcgcctggtt taaacgcaac accttaaata
     3721 aacccgtgat tatgggccgc catacctggg aatcaatcgg tcgtccgttg ccaggacgca
     3781 aaaatattat cctcagcagt caaccgggta cggacgatcg cgtaacgtgg gtgaagtcgg
     3841 tggatgaagc catcgcggcg tgtggtgacg taccagaaat catggtgatt ggcggcggtc
     3901 gcgtttatga acagttcttg ccaaaagcgc aaaaactgta tctgacgcat atcgacgcag
     3961 aagtggaagg cgacacccat ttcccggatt acgagccgga tgactgggaa tcggtattca
     4021 gcgaattcca cgatgctgat gcgcagaact ctcacagcta ttgctttgag attctggagc
     4081 ggcgggtcga cagtaaagga gaagaacttt tcactggaat ggtgagcaag ggcgaggagg
     4141 ataacatggc ctctctccca gcgacacatg agttacacat ctttggctcc atcaacggtg
     4201 tggactttga catggtgggt cagggcaccg gcaatccaaa tgatggttat gaggagttaa
     4261 acctgaagtc caccaagggt gacctccagt tctccccctg gattctggtc cctcatatcg
     4321 ggtatggctt ccatcagtac ctgccctacc ctgacgggat gtcgcctttc caggccgcca
     4381 tggtagatgg ctccggatac caagtccatc gcacaatgca gtttgaagat ggtgcctccc
     4441 ttactgttaa ctaccgctac acctacgagg gaagccacat caaaggagag gcccaggtga
     4501 aggggactgg tttccctgct gacggtcctg tgatgaccaa ctcgctgacc gctgcggact
     4561 ggtgcaggtc gaagaagact taccccaacg acaaaaccat catcagtacc tttaagtgga
     4621 gttacaccac tggaaatggc aagcgctacc ggagcactgc gcggaccacc tacacctttg
     4681 ccaagccaat ggcggctaac tatctgaaga accagccgat gtacgtgttc cgtaagacgg
     4741 agctcaagca ctccaagacc gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg
     4801 tgatgggcat ggacgagctg tacaagaaga agctttaccc atacgatgtt ccagattacg
     4861 ctacgcgttt ccccaacaat atccagatag gggggttatt tccaaaccaa caatcacagg
     4921 aacatgcggc ttttaggttt gctttgtcac aactcacgga gccccccaag ctgcttcccc
     4981 agatcgatat tgtgaacatc agcgacagct ttgagatgac ttaccgtttc tgttcccagt
     5041 tctccaaagg agtctatgcc atctttggat tttatgaacg aaggactgtc aacatgctga
     5101 cctccttctg tggggccctc catgtgtgct tcattactcc aagttttcct gttgacacat
     5161 ccaatcaatt tgtccttcag ctacgcccgg aactacagga agctctcatt agcattatcg
     5221 accattacaa gtggcaaacc tttgtctaca tttatgatgc tgaccggggc ctgtcagtcc
     5281 tgcagagagt cttggataca gccgcagaga agaactggca ggtaacggct gtcaacattc
     5341 tgacaaccac cgaggaagga taccggatgc tctttcagga cctggagaag aaaaaggaga
     5401 ggctggtggt ggttgactgt gaatcagaac gcctcaacgc catcctgggc cagatcgtga
     5461 agctagaaaa gaatggcatc gggtaccact acatcctcgc caatctgggc ttcatggaca
     5521 ttgacttaaa taagttcaag gagagcggag ccaatgtgac aggtttccag ctggtgaact
     5581 acacagacac gatcccagcc agaatcatgc agcaatggag gacaagtgac tcccgagacc
     5641 ataccagggt ggactggaag aggccaaagt acacttctgc tctcacctat gatggtgtca
     5701 aggtgatggc tgaggccttc caaagcctgc ggaggcagag gattgacata tcccgccggg
     5761 ggaatgctgg ggactgtctg gctaacccag ctgtgccctg gggtcaaggg atcgacatcc
     5821 agagagccct gcagcaggtg cgcttcgaag gtttgacagg aaatgtgcag ttcaacgaga
     5881 aagggcgccg gaccaattac accctccacg tgatcgaaat gaaacatgat ggaatccgaa
     5941 agattggtta ctggaatgaa gacgataaat ttgtccccgc agccaccgac gctcaggctg
     6001 gaggggacaa ctcaagcgtc cagaatagga cctacatcgt cactactatc ctcgaagatc
     6061 cttacgtgat gcttaaaaag aatgccaacc agtttgaggg caatgaccgc tatgagggct
     6121 actgtgtgga gctggctgca gagatcgcca agcacgtggg ctactcctac cgacttgaga
     6181 ttgtcagcga cggcaaatat ggagcccggg atcccgacac aaaggcttgg aatggcatgg
     6241 tgggagaact ggtctatgga agagcagacg tggctgtggc tcccttgacc ataaccttgg
     6301 tccgggagga agtcatcgac ttctccaagc cattcatgag tttgggaatc tccattatga
     6361 ttaagaagcc acagaagtcc aagccaggtg tcttctcctt tcttgaccct ttggcctatg
     6421 agatctggat gtgtatagtg tttgcctaca ttggagtgag cgtcgtcctc ttcctggtca
     6481 gccgtttcag cccctacgaa tggcacagcg aagagtttga agagggacga gaccagacaa
     6541 ccagtgacca gtcaaatgag tttggcatat tcaacagcct gtggttctcc ctgggggcct
     6601 tcatgcagca aggatgtgac atttccccca ggtccctgtc cggacgcatc gtcggcggcg
     6661 tctggtggtt cttcactttg atcatcatct cctcgtacac agccaacctg gctgccttcc
     6721 tgactgtgga gaggatggtg tctcccattg agagtgcaga ggacctggca aagcagacgg
     6781 aaattgctta tgggacattg gaagcaggct ccactaagga gttcttcagg agatctaaaa
     6841 tcgctgtgtt tgagaagatg tggacataca tgaagtctgc agaaccatcc gtgtttgttc
     6901 ggaccacaga ggaaggcatg atcagagtga gaaaatctaa aggcaaatac gcctacctcc
     6961 tggagtccac catgaatgag tatattgagc aacgaaagcc ctgtgacacc atgaaagtgg
     7021 gaggtaactt ggattccaaa ggctatggca ttgcgacacc caaggggtcc gccctgagaa
     7081 atccagtaaa cctggcagtg ttaaaactga acgagcaggg gcttttggac aaattgaaaa
     7141 acaaatggtg gtacgacaag ggcgagtgcg gcagcggggg aggtgactcc aaggacaaga
     7201 ccagcgcttt gagcctcagc aatgtggcag gcgtgttcta catcctgatt ggagggctgg
     7261 gactggccat gctggttgcc ttaatcgagt tctgctacaa atcccgtagc gagtcgaagc
     7321 ggatgaaggg tttctgtttg atcccacagc aatccatcaa tgaagccata cggacatcga
     7381 ccctcccccg gaacagtggg gcaggagcca gcggaggagg cggcagtgga gagaatggcc
     7441 gggtggtcag ccaggacttc cccaagtcca tgcaatccat tccctgcatg agtcacagtt
     7501 cagggatgcc cttgggagcc acaggattgt aagaattctg cagtcgacgg taccgcgggg
     7561 tacaagtaaa gcggccgcaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg
     7621 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     7681 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     7741 ttttcattgc aatagtgtgt tggaattttt tgtgtctatc actcggaagg acatatggga
     7801 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     7861 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     7921 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     7981 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     8041 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     8101 ggagatccct cgacctgcac cgtcgaccag ctggtcgacg gtgcaccgtc gaccagcttg
     8161 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     8221 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     8281 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     8341 atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     8401 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     8461 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     8521 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     8581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8641 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     8701 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     8761 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     8821 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccag
//
