LOCUS       sequence-199106-        8305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of PDZD8, sh resistant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8305)
  AUTHORS   Hirabayashi Y, Kwon SK, Paek H, Pernice WM, Paul MA, Lee J, Erfani 
            P, Raczkowski A, Petrey DS, Pon LA, Polleux F
  TITLE     ER-mitochondria tethering by PDZD8 regulates Ca(2+) dynamics in 
            mammalian neurons.
  JOURNAL   Science. 2017 Nov 3;358(6363):623-630. doi: 10.1126/science.aan6009.
  PUBMED    29097544
REFERENCE   2  (bases 1 to 8305)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2017-11-3- 3"; volume: "358"; issue: 
            "6363"; pages: "623-630"
COMMENT     Sequence Label: pCAG-PDZD8HA_sh_resistant
FEATURES             Location/Qualifiers
     source          1..8305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(42..629)
                     /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(800..1591,1592..1660))
                     /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     1423..1442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1661..1765)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1796..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"
     CDS             6969..6995
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(7089..7108)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7154..7209
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7208..7227)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7600..7616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7600..7616)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7613..7635)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7624..7640
                     /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(7648..7654,7655..7672,7673..7678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7693..7714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7773..7968
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      7819..7954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7974..8108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8011..8030)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8065..8084
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8176..8193)
                     /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 agacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
      781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
      841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat
     1801 tgattattga ctagttatta 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 gggctgctgc
     3541 tcctgatcct ggcgtcggcc gtgctgggct cgttcctcac gctgctcgca cagttcctgc
     3601 tactctaccg cagacagccc gagccgcggg cggacgaagc ggcccgcgcg ggcgacggct
     3661 tccgctacct taagccggtg ccgggcctgc ccctcaggga gtacctttat ggcggcggcg
     3721 ccgaggagct cgctgcttgc tcctccgagg ccggcgccag ctcgaccccg acccccgaca
     3781 gcccggcccc gccgacgctg gagacctgct acttcctcaa cgccaccatc ctgttcctgt
     3841 tccgggagct gcgggacacc gcgctcgccc gccgctgggt caccaagaag atcaaggtgg
     3901 agttcgagga gctgctgcag accaagacgg ccggccgcct gctggagggg ctgagtctgc
     3961 gcgacgtgtt cctgggcgac acggtgccct tcatcaagac catccggctg gtgcggcccg
     4021 tggtggcctc gggcaccggc gagcccgacg accccgacgg ggacgcgctg cccgccacct
     4081 gcccggagga gctggccttt gaagcggagg tggagtacaa cggcggtttc cacctggcca
     4141 tcgacgtgga cctggtgttt ggcaagtccg cctacctgtt cgtgaagctg tctcgcgtgg
     4201 tggggaggct gcgcttcgtc ctcacccgcg tgcccttcac ccactggttc ttctccttcg
     4261 tggaggaccc gctgattgac ttcgaggtgc gctcccagtt cgaggggcgg cccatgcccc
     4321 agctcacctc catcatcgtc aaccagctca agaagatcat caagcgcaag cacaccctgc
     4381 ccagttacaa gatcaggttt aagccgtttt ttccatacca agccttgcaa ggatttgaag
     4441 aagatgaaga acttatccat attcaacaat gggcacttac tgaaggccgg cttaaagtta
     4501 cattgttaga atgtagcagg ttattcattt ttggatccta tgacagagaa acaaatgttc
     4561 attgcacact tgagctgagc agtggtgttt gggaagaaaa acaaaggagt tctattaaga
     4621 cggttgaatt aataaaaggg aatttacaaa gtgtcggact tacacttcgt cttgttcagt
     4681 caactgatgg gtatgctggg catgtcataa tcgaaactgt ggccccaaac tcacctgctg
     4741 cgatggcaga tcttcaacgg ggagatcggc tcattgctat tggaggtgtg aaaatcacat
     4801 cgaccctgca ggtgctgaag ctcatcaagc aggctgggga ccgcgtgctg gtgtactacc
     4861 agaggccggc tgggcagagc agccaagaca gcctgggcca gttggaagaa agcttcttgt
     4921 ccagctcctg ccaggcagcc tacgaagagg acgctgctgg gctgtcagcg gacactgaaa
     4981 acagagacct agattctgaa ttcgaagact tggcgagtga tgtcagagta cagacagagt
     5041 tgaaagagga gacacagccg ttgagtcaca gtcccaaacg tactccaaca acactttcta
     5101 ttaaacccct tggcgctata tccccggttt taaatcgtaa attaatttca ggaatccacc
     5161 caccaccaca aaaacttcca tccaaagaag gaaataaacc atcaactcta aaaacttcag
     5221 agaccacaga agcagcacaa gtgtccaaac cccaaggacc cactttcaaa ccacctgtgc
     5281 caccgcgacc acaggggaga gtgcctctgc ctcccactga cacctcagcc caagcagacc
     5341 cggaagcccc agaaaagcca gacaaggtgc tgctccctcc tcctcctgca gataaacctg
     5401 ccgagaagca agtgaagagt gtggatcaag gagaagatgt ggcagcaggt aagcagtctt
     5461 cagcgaagca ggaaggagtg aaagatcttc cttcagaaag ttctgctcct actaaagact
     5521 cttcagatga ccctcagatg tgggaatcct cagaggttct atatcgcaac aaggtgggaa
     5581 agtggtcaag aaccagagcg tcctgtgtgt ttgacataga agcctgccat aggtacctga
     5641 acatcgcact gtggtgcagg gatcctttca agttaggagg tctcatctgt ttggggcatg
     5701 ttagtttaaa acttgaggaa gtggctttag gatgtctagc tacatcaaac atggaatacc
     5761 ttacaaagtt cagactggaa ccccctacac ctaaggccat ggtcactaga actgcactac
     5821 gtaatctgag tatgcaaaag ggctttaatg acaaattttg ttttggtgac attactattc
     5881 atttcaagta tttgaaggaa ggagaaccag accaccacat agttcctaat gtagagaaag
     5941 aaaaagaact ccatttggtt gaggaggttt ctacactccc taaagaagag cattttgtgg
     6001 gacagatgag tttgtcagaa aataaacata gtttccaaga tactcagttc cagaacccaa
     6061 cttggtgtga ttactgtaag aaaaaagttt ggacaaaagc tgcttcccag tgtatgttct
     6121 gtgcttatgt ttgtcataaa aaatgtcagg aaaagtgtct agctgagaca cctctttgtg
     6181 gagcaactga gcgacggata gaccggaccc tgaaaaacct caggctggaa ggacaggacc
     6241 cactcttggg ccttcctcct cgtgttgaga tagaagctaa caagtcagtc aataaaacaa
     6301 cagggttgac aaggcacatc attaatacga gctctcgttt actaaatctg cgtcaagtct
     6361 ctaaaactcg cctttctgaa ccaggaacag atcttgtaga accttcacca aaacacacac
     6421 ccaatacgtc agacaatgaa ggcagtgaca cagaggtctg tggttcaaac agtccttcca
     6481 aacgaggaaa cagtgcagga ataaagttaa tgagaaagga aggagggctg gatgacagtg
     6541 ttttcattgc agtaaaggaa attggtcgtg atctgtacag aggcttgccg acagaagaga
     6601 ggatccagaa gctggagttc atgttggata agctacagaa tgaaattgat caggagctgg
     6661 aacacaacaa ttcccttgtt agagaagaaa aggaaacaaa tgatacaagg aaaaagtcag
     6721 ttctttctgc tgctttagct aaatcgggag aaagactaca agctctgaca cttctgatga
     6781 tccactacag agcgggaatc gaagatattg aaaccctaga aaatctgtct ttagaccagc
     6841 actcgaaaaa gatgaacaag tacgcagatg atacagaaga agacctcgat agtgagataa
     6901 gccagttgat agactctcag ccattcagca acatttccga tgacttgttc ggcccatctg
     6961 agtcagtgta cccatacgat gttccagatt acgcttaagc ggccgcaatt cactcctcag
     7021 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
     7081 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     7141 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     7201 tgtctatcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     7261 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     7321 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     7381 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     7441 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     7501 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcaccg tcgaccagct
     7561 ggtcgacggt gcaccgtcga ccagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7621 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7681 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7741 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     7801 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     7861 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     7921 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     7981 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     8041 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     8101 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     8161 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     8221 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     8281 aaagaacatg tgagcaaaag gccag
//
