LOCUS       Exported                4958 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Longitudinal monitoring of Gaussia and Nano luciferase activities to
            simultaneously assess ER calcium homeostasis and ER stress in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    pOTTC840 - pAAV 5xATF6 secNLuc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4958)
  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 4958)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4958
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             523..1035
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     misc_feature    1085..1673
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1138..1158)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1556..1567)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1705..2181
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1904..1923)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2221..2361
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2221..2350
                     /label=AAV2 ITR
     rep_origin      2436..2891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2523..2542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2733..2754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2908..2927)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3027..3049
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3087..3105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3173..3277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3278..4138
                     /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(3496..3515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4309..4897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4798..4817
                     /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 gcgttgcagc ccaagcttgc tcgagacagg tgctgacgtg
      181 gcgattcctc gagacaggtg ctgacgtggc gattcctcga gacaggtgct gacgtggcga
      241 ttcctcgaga caggtgctga cgtggcgatt cctcgagaca ggtgctgacg tggcgattcc
      301 tcgagatctg cactcattca taaaacgctt gttataaaag cagtggctgc ggcgcctcgt
      361 actccaaccg catctgcagc gagcaactga gaagccaaga ctgagccggc ggccggtact
      421 gttggtaaag ccaccatgaa ctccttctcc acaagcgcct tcggtccagt tgccttctcc
      481 ctgggcctgc tcctggtgtt gcctgctgcc ttccctgccc cagtcttcac actcgaagat
      541 ttcgttgggg actggcgaca gacagccggc tacaacctgg accaagtcct tgaacaggga
      601 ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc
      661 ctgagcggtg aaaatgggct gaagatcgac atccatgtca tcatcccgta tgaaggtctg
      721 agcggcgacc aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat
      781 catcacttta aggtgatcct gcactatggc acactggtaa tcgacggggt tacgccgaac
      841 atgatcgact atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc
      901 actgtaacag ggaccctgtg gaacggcaac aaaattatcg acgagcgcct gatcaacccc
      961 gacggctccc tgctgttccg agtaaccatc aacggagtga ccggctggcg gctgtgcgaa
     1021 cgcattctgg cgtaaggccg cgactctaga gtggcgcgcc gaattcgata tcaagcttat
     1081 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     1141 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     1201 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     1261 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     1321 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     1381 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     1441 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     1501 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     1561 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     1621 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     1681 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     1741 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     1801 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     1861 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     1921 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     1981 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     2041 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     2101 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     2161 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     2221 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2281 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     2341 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     2401 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     2461 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2521 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2581 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2641 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2701 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2761 actcaaccct atctcgggct attcttttga tttataaggg attttgccga tttcggccta
     2821 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2881 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2941 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3001 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     3061 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     3121 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     3181 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3241 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3301 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3361 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3421 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3481 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3541 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3601 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3661 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3721 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3781 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3841 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3901 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3961 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     4021 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     4081 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     4141 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4201 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4261 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4321 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4381 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4441 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4501 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4561 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4621 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4681 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4741 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4801 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4861 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4921 acggttcctg gccttttgct ggccttttgc tcacatgt
//
