LOCUS       V007427                 8341 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V007427
VERSION     V007427
KEYWORDS    pLYS1-FLAG-MitoGFP-HA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8341)
  AUTHORS   Sancak Y, Markhard AL, Kitami T, Kovacs-Bogdan E, Kamer KJ, Udeshi
            ND, Carr SA, Chaudhuri D, Clapham DE, Li AA, Calvo SE, Goldberger O,
            Mootha VK
  TITLE     EMRE is an Essential Component of the Mitochondrial Calcium
            Uniporter Complex.
  JOURNAL   Science. 2013 Nov 14.
   PUBMED   24231807
REFERENCE   2  (bases 1 to 8341)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8341)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science.
            2013 Nov 14."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1073..1306
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1491..1535
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1684..1725
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     primer_bind     1833..1853
                     /label="hU6-F"
                     /note="Human U6 promoter, forward primer"
     promoter        2142..2345
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2560..2583
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2614..3330
                     /label="EGFP"
                     /note="enhanced GFP"
     CDS             3358..3384
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    3435..3552
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        3601..4111
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4133..4729
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             4860..5093
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5165..5299
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5326..5461
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(5482..5500)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5510..5526)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      5668..6123
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6149..6253
                     /label="AmpR promoter"
     CDS             6254..7111
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      7285..7873
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     8027..8044
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     protein_bind    8161..8182
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8197..8227
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    8235..8251
                     /label="lac operator"
                     /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)."
     primer_bind     8259..8275
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        8296..8314
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1741 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgatcac
     1801 gagactagcc tcgagcggcc gcccccttca ccgagggcct atttcccatg attccttcat
     1861 atttgcatat acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca
     1921 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     1981 ttttaaaatt atgttttaaa atggactatc atatgccaag tacgccccct attgacgtca
     2041 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2101 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     2161 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     2221 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     2281 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     2341 gagctggttt agtgaaccgt cagatccgct agcatggcgg ccgccgcagg tagatcgctc
     2401 ctgctgctcc tctcctctcg gggcggcggc ggcgggggcg ccggcggctg cggggcgctg
     2461 actgccggct gcttccctgg gctgggcgtc agccgccacc ggcagcagca gcaccaccgg
     2521 acggtacacc agaggatcgc ttccggaggc accggtatgg attacaagga tgacgatgac
     2581 aagggtggat ctggtggatc tggtggaccc gggatggtga gcaagggcga ggagctgttc
     2641 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2701 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     2761 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2821 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2881 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2941 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     3001 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     3061 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     3121 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     3181 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     3241 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     3301 atcactctcg gcatggacga gctgtacaag ggtggatctg gtggatctgg tggatcttac
     3361 ccctatgatg tgcctgacta cgcataaggt accgaattct cgacctcgag acaaatggca
     3421 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     3481 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     3541 attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag
     3601 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc
     3661 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc
     3721 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc
     3781 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac
     3841 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc
     3901 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag
     3961 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct
     4021 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct
     4081 cgttgaccga atcaccgacc tctctcccca gggggatcca ccggagctta ccatgaccga
     4141 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     4201 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     4261 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     4321 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     4381 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     4441 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     4501 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     4561 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     4621 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     4681 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     4741 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcgg tacctttaag
     4801 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     4861 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     4921 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     4981 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     5041 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     5101 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     5161 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5221 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5281 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact
     5341 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5401 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5461 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     5521 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5581 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5641 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     5701 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5761 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5821 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5881 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5941 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6001 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     6061 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     6121 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6181 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6241 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6301 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6361 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6421 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6481 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6541 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6601 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6661 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6721 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6781 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6841 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6901 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6961 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7021 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7081 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7141 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7201 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7261 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7321 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7381 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7441 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7501 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7561 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7621 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7681 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7741 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7801 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7861 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7921 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7981 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     8041 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     8101 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     8161 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     8221 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     8281 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta
     8341 a
//
