LOCUS       sequence-384552-        7395 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ASAP3ER under the control of the pCAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7395)
  AUTHORS   Campbell EP, Abushawish AA, Valdez LA, Bell MK, Haryono M, Rangamani
            P, Bloodgood BL
  TITLE     Electrical signals in the ER are cell type and stimulus specific 
            with extreme spatial compartmentalization in neurons.
  JOURNAL   Cell Rep. 2023 Jan 31;42(1):111943. doi: 
            10.1016/j.celrep.2022.111943. Epub 2023 Jan 5.
  PUBMED    36640310
REFERENCE   2  (bases 1 to 7395)
  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: "Cell Rep.";
            date: "2023-01-31"; volume: "42(1):111943. doi"; pages: " 
            10.1016/j.celrep.2022.111943. Epub 2023 Jan 5"
COMMENT     Sequence Label: pCAG-ASAP3ER
FEATURES             Location/Qualifiers
     source          1..7395
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      14..23
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1424..1441)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1430..1654
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1700..2128
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1787..1806)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1997..2018
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2137..2157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2142..2471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2322..2457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2384..2403
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2482..3081
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2482..2501)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2978..2998
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3241..3362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3278..3297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3332..3351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3411..3427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3411..3427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3424..3446)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3435..3451
                     /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(3459..3465,3466..3483,3484..3489))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3504..3525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3642..3659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3813..4401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3893..3912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4572..5363,5364..5432))
                     /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     5195..5214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5433..5537)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5569..5948
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6227..7243
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7167..7189
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7251..7270
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gctagctgta cacgccacca tggagacgac tgtgaggtat gaacaggggt cagagctcac
       61 taaaacttcg agctctccaa cagcagatga gcccacgata aagattgatg atggtcgtga
      121 tgagggtaat gaacaagaca gctgttccaa taccattagg agaaaaattt ccccgtttgt
      181 gatgtcattt ggattcagag tatttggagt tgtgcttatc attgtagaca tcatagtggt
      241 gattgtggat ctggccatca gtgagaagaa aagaggcatt agagagattc ttgaaggtgt
      301 ttccctggct atagcactct tcttccttgt tgatgttctc atgagagtgt ttgttgaagg
      361 cttcaagaac tatttccggt ccaaactgaa tactttggat gcagtcatag tagtgggcac
      421 tctgctaatt aatatgacct actccttctc tgacggtaca tttcgcggtg ataacgtgta
      481 tattaccgcg gataaacaga aaaacggcat taaagcgaac tttaccgtgc gccataacgt
      541 ggaagatggc agcgtgcagc tggcggatca ttatcagcag aacaccccga ttggcgatgg
      601 cccggtgctg ctgccggata accattatct gagcacccag accgtgctga gcaaagatcc
      661 gaacgaaaaa cgcgatcaca tggtgctgct ggaatttgtg accgcagcgg gcattacaca
      721 cggcatggat gaactgtatg gcggcactgg cggcagcgcg agccagggcg aagaactgtt
      781 taccggcgtg gtgccgattc tggtggaact ggatggcgat gtgaacggcc ataaatttag
      841 cgtgcgcggc gaaggcgaag gcgatgcgac cattggcaaa ctgaccctga aatttatttg
      901 caccaccggc aaactaccgg tgccgtggcc gaccctggtg accaccttaa cctatggcgt
      961 gcagtgcttt agccgctatc cggatcatat gaaacgccat gattttttta aaagcgcgat
     1021 gccggaaggc tatgtgcagg aacgcaccat tagctttaaa gatgatggca aatataaaac
     1081 ccgcgcggtg gtgaaatttg aaggcgatac cctggtgaac cgcattgaac tgaaaggcac
     1141 cgattttaaa gaagatggca acattctggg gcataaactg gaatataaca cagatcagat
     1201 gccgcagatg gttactcttc ttcgagttct gagaattgtt atcttaataa gaatatttca
     1261 gctggcttca cagaagaaac aacttgaagt ggtaaccaag aaggcggcgt aaaagcttgg
     1321 taccgagctc ggatccacta gtccagtgtg gtggaattct gcagatatcc agcacagtgg
     1381 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     1441 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1501 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1561 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1621 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     1681 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1741 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1801 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1861 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1921 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     1981 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2041 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2101 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2221 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2281 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2341 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2401 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2461 gcttttgcaa aaagctcccg catgaccgag tacaagccca cggtgcgcct cgccacccgc
     2521 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     2581 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     2641 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     2701 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     2761 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     2821 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     2881 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     2941 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     3001 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     3061 acccgcaagc ccggtgcctg attcgaaatg accgaccaag cgacgcccaa cctgccatca
     3121 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     3181 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     3241 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3301 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3361 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     3421 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     3481 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3541 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     3601 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     3661 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     3721 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     3781 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     3841 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     3901 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     3961 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4021 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4081 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4141 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4201 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4261 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4321 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4381 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4441 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4501 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4561 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4621 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4681 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4741 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4801 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4861 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4921 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4981 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5041 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5101 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5161 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5221 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5281 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5341 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5401 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5461 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5521 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cattgattat
     5581 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     5641 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     5701 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     5761 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     5821 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     5881 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     5941 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     6001 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     6061 gggggggnnn sgsgcgccrg gsgggggggg ggggggsggg gggsggggsg gggsgrggcg
     6121 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
     6181 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
     6241 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
     6301 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
     6361 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
     6421 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
     6481 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
     6541 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
     6601 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
     6661 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
     6721 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
     6781 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
     6841 cggggrgggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
     6901 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     6961 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     7021 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     7081 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     7141 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     7201 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     7261 ggttattgtg ctgtctcatc attttggcaa agaattcctc gaggaattca ctcctcaggt
     7321 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     7381 ctgagatctg catga
//
