LOCUS       Exported                8565 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses dsRed and scramble shRNA under the control of the CMV 
            promoter in eukaryotic cells.
ACCESSION   .
VERSION     .
KEYWORDS    pPRIME-dsRed-shscramble
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8565)
  AUTHORS   Yu X, Ye Z, Houston CM, Zecharia AY, Ma Y, Zhang Z, Uygun DS, Parker
            S, Vyssotski AL, Yustos R, Franks NP, Brickley SG, Wisden W
  TITLE     Wakefulness Is Governed by GABA and Histamine Cotransmission.
  JOURNAL   Neuron. 2015 Jun 17. pii: S0896-6273(15)00516-4. doi: 
            10.1016/j.neuron.2015.06.003.
  PUBMED    26094607
REFERENCE   2  (bases 1 to 8565)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8565
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1023..1148
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1645..1878
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2063..2107
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2405..2522
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2659..2962
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2963..3166
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3212..3889
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MDSSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(3342..3360)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3392..3412)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3610..3629
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3804..3827
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     ncRNA           3939..4060
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4087..4101
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           4128..4254
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        4408..4510
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4511..5170
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4577..4596)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5302..5331
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    5454..6042
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5507..5527)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5925..5936)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6045..6061)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6046..6062)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6571..6751
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6813..7401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6893..6912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7572..8432)
                     /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     8195..8214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8433..8537)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata gaccagatct gagcctggga gctctctggc
      841 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      901 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      961 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     1021 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     1081 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     1141 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     1201 cagggggaaa gaaaaaatat aaataaacat atagtatggg caagcaggga gctagaacga
     1261 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     1321 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     1381 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     1441 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgg ccgcgctgat
     1501 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1561 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1621 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1681 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     1741 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     1801 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     1861 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     1921 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     1981 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     2041 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     2101 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     2161 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     2221 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2281 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     2341 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
     2401 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2461 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2521 ttcgggttta ttacagggac agcagagatc cagtttggtt agtaccgggc ccgctctaga
     2581 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     2641 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2701 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2761 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2821 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2881 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2941 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     3001 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     3061 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     3121 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     3181 tcagatccgc tagcgctacc gggcctgcag gatggactcc tccgaggacg tcatcaagga
     3241 gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac ggccacgagt tcgagatcga
     3301 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     3361 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttccagt acggctccaa
     3421 ggtgtacgtg aagcaccccg ccgacatccc cgactacaag aagctgtcct tccccgaggg
     3481 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     3541 ctcctccctg caggacggct ccttcatcta caaggtgaag ttcatcggcg tgaacttccc
     3601 ctccgacggc cccgtaatgc agaagaagac tatgggctgg gaggcctcca ccgagcgcct
     3661 gtacccccgc gacggcgtgc tgaagggcga gatccacaag gccctgaagc tgaaggacgg
     3721 cggccactac ctggtggagt tcaagtccat ctacatggcc aagaagcccg tgcagctgcc
     3781 cggctactac tacgtggact ccaagctgga catcacctcc cacaacgagg actacaccat
     3841 cgtggagcag tacgagcgcg ccgagggccg ccaccacctg ttcctgtagg cggccgcaag
     3901 ccttgttaag tgctcgcttc ggcagcacat atactatgtt gaatgaggct tcagtacttt
     3961 acagaatcgt tgcctgcaca tcttggaaac acttgctggg attacttctt caggttaacc
     4021 caacagaagg ctcgagaagg tatattgctg ttgacagtga gcggccgcga ttaggctgtt
     4081 ataatagtga agccacagat gtattataac agcctaatcg cggctgccta ctgcctcgga
     4141 attcaagggg ctactttagg agcaattatc ttgtttacta aaactgaata ccttgctatc
     4201 tctttgatac atttttacaa agctgaatta aaatggtata aattaaatca cttttttcaa
     4261 ttggaagact aatgcgttta aacacgcggc gacgcgttcg accgaataaa tacctgtgac
     4321 ggaagatcac ttcgcagaat aaataaatcc tggtgtccct gttgataccg ggaagccctg
     4381 ggccaacttt tggcgaaaat gagacgttga tcggcacgta agaggttcca actttcacca
     4441 taatgaaata agatcactac cgggcgtatt ttttgagttg tcgagatttt caggagctaa
     4501 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     4561 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     4621 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     4681 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattac gtatggcaat
     4741 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     4801 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     4861 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     4921 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     4981 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     5041 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     5101 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     5161 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     5221 aataagtgat aataagcgga tgaatggcag aaattcggat ctcgaccgcg tttgggcggt
     5281 ggctccctgc cacgcggctc cgaacagaag ctgatctccg aagaggatct gacatgtgtt
     5341 taaacctcga cttaattaag tcgagggtcg acggtatcga taagctcgct tcacgagatc
     5401 atgtttaagg gttccggttc cactaggtac aattcgatat caagcttatc gataatcaac
     5461 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5521 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5581 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5641 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5701 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5761 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5821 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     5881 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5941 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6001 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     6061 gatcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     6121 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     6181 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     6241 aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     6301 atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc
     6361 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta
     6421 gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg
     6481 gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac
     6541 atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc
     6601 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6661 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6721 cccttttagt cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc
     6781 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6841 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6901 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6961 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7021 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7081 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7141 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7201 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7261 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7321 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7381 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7441 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7501 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7561 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7621 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7681 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7741 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7801 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7861 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7921 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7981 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8041 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8101 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8161 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8221 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8281 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8341 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8401 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8461 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8521 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac
//
