LOCUS       sequence-492036-        4792 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Longitudinal monitoring of Gaussia and Nano luciferase activities to
            simultaneously assess ER calcium homeostasis and ER stress in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4792)
  AUTHORS   Wires ES, Henderson MJ, Yan X, Back S, Trychta KA, Lutrey MH, Harvey
            BK
  TITLE     Longitudinal monitoring of Gaussia and Nano luciferase activities to
            concurrently assess ER calcium homeostasis and ER stress in vivo.
  JOURNAL   PLoS One. 2017 Apr 12;12(4):e0175481. doi: 
            10.1371/journal.pone.0175481. eCollection 2017.
  PUBMED    28403212
REFERENCE   2  (bases 1 to 4792)
  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: "PLoS One.";
            date: "2017-04-12"; volume: "12(4):e0175481. doi"; pages: " 
            10.1371/journal.pone.0175481. eCollection 2017"
COMMENT     Sequence Label: pOTTC841 - pAAV minP secNLuc
FEATURES             Location/Qualifiers
     source          1..4792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             357..869
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     misc_feature    919..1507
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(972..992)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1390..1401)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1539..2015
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1738..1757)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2055..2195
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2055..2184
                     /label=AAV2 ITR
     rep_origin      2270..2725
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2357..2376)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2567..2588
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2742..2761)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2861..2883
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2921..2939)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3007..3111
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3112..3180,3181..3972)
                     /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     complement(3330..3349)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4143..4731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4632..4651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcactca ttcataaaac gcttgttata aaagcagtgg
      181 ctgcggcgcc tcgtactcca accgcatctg cagcgagcaa ctgagaagcc aagactgagc
      241 cggcggccgg tactgttggt aaagccacca tgaactcctt ctccacaagc gccttcggtc
      301 cagttgcctt ctccctgggc ctgctcctgg tgttgcctgc tgccttccct gccccagtct
      361 tcacactcga agatttcgtt ggggactggc gacagacagc cggctacaac ctggaccaag
      421 tccttgaaca gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc gtaactccga
      481 tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc
      541 cgtatgaagg tctgagcggc gaccaaatgg gccagatcga aaaaattttt aaggtggtgt
      601 accctgtgga tgatcatcac tttaaggtga tcctgcacta tggcacactg gtaatcgacg
      661 gggttacgcc gaacatgatc gactatttcg gacggccgta tgaaggcatc gccgtgttcg
      721 acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt atcgacgagc
      781 gcctgatcaa ccccgacggc tccctgctgt tccgagtaac catcaacgga gtgaccggct
      841 ggcggctgtg cgaacgcatt ctggcgtaag gccgcgactc tagagtggcg cgccgaattc
      901 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
      961 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     1021 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     1081 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     1141 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     1201 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     1261 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     1321 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc
     1381 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     1441 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     1501 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct
     1561 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     1621 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     1681 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     1741 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     1801 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     1861 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     1921 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     1981 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg
     2041 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     2101 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     2161 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     2221 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     2281 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2341 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2401 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2461 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     2521 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2581 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     2641 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     2701 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     2761 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     2821 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     2881 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     2941 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3001 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3061 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3121 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3181 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3241 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3301 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3361 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3421 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3481 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3541 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3601 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3661 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3721 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3781 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3841 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3901 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3961 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4021 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4081 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4141 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4201 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4261 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4321 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4381 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4441 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4501 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4561 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4621 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4681 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4741 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
