LOCUS       sequence-86304-e        7646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Dre in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7646)
  AUTHORS   Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone
            J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H,
            Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, 
            Deisseroth K
  TITLE     Targeting cells with single vectors using multiple-feature Boolean 
            logic.
  JOURNAL   Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 
            2014 Jun 8.
  PUBMED    24908100
REFERENCE   2  (bases 1 to 7646)
  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: 
            "10.1038/nmeth.2996"; journalName: "Nat Methods"; date: "2014-07"; 
            volume: "11"; issue: "7"; pages: "763-72"
COMMENT     Sequence Label: pAAV-EF1a-mCherry-IRES-Dre
FEATURES             Location/Qualifiers
     source          1..7646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1432..2139
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1574..1592)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1842..1861
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2156..2708
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2300..2317)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2527..2546
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3773..4361
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3826..3846)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4244..4255)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4393..4869
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4592..4611)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4909..5049
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4909..5038
                     /label=AAV2 ITR
     rep_origin      5124..5579
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5211..5230)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5421..5442
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5596..5615)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5715..5737
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5775..5793)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5861..5965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5966..6034,6035..6826)
                     /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(6184..6203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6997..7585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7486..7505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgtgagcaag
     1441 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     1501 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     1561 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     1621 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     1681 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1741 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     1801 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     1861 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     1921 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     1981 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     2041 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     2101 cgccactcca ccggcggcat ggacgagctg tacaagtaag gcgcgccccc ccctaacgtt
     2161 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     2221 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     2281 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     2341 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     2401 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     2461 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     2521 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     2581 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     2641 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     2701 tgataatatg gccacaacca tgtcagagct tataatttca ggtagtagtg gaggatttct
     2761 cagaaacatt ggcaaggagt accaagaagc tgccgaaaat ttcatgaggt ttatgaacga
     2821 ccagggcgca tacgctccaa atactttgag agacctccga ctcgtgttcc acagctgggc
     2881 tcgatggtgc cacgccaggc agcttgcatg gtttccaata tctccagaaa tggctcgcga
     2941 gtactttttg caactccacg acgctgatct ggcatctacc accatcgaca agcactatgc
     3001 aatgctgaat atgctgctgt cccactgtgg actgcctcca ctgtcagatg acaagtccgt
     3061 gagcctggcg atgcggagaa ttagaagaga ggcggccact gaaaaggggg aacggacagg
     3121 acaggcgata cctctccgct gggacgacct gaaactgctg gacgtactcc tgagtcggag
     3181 cgagcgcttg gtcgatcttc ggaaccgcgc cttcctcttc gtggcctata ataccctgat
     3241 gcggatgagt gagatcagca gaatccgagt tggggatctg gatcagaccg gggacactgt
     3301 gactttgcat atctcccata ctaagactat caccaccgct gccggcttgg acaaggttct
     3361 ctctaggcgc acgaccgctg tcctgaatga ctggcttgat gtatctggcc tgcgagaaca
     3421 tcccgacgcc gtgctgttcc cccctattca ccggtctaac aaggcaagaa tcacgacaac
     3481 gccgctgact gcacccgcca tggagaagat cttttccgat gcttgggtgc tccttaacaa
     3541 aagggatgca acacctaaca aaggtagata caggacatgg accgggcata gcgcgcgggt
     3601 gggggccgcg attgatatgg ccgagaaaca ggtcagcatg gtggagatca tgcaggaggg
     3661 tacatggaaa aaaccggaga cactgatgag atatcttagg agggggggcg tcagcgttgg
     3721 agcaaatagc cgcctgatgg acagttgaga attcgatatc aagcttatcg ataatcaacc
     3781 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3841 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3901 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3961 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     4021 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     4081 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     4141 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     4201 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     4261 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     4321 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     4381 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     4441 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     4501 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     4561 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     4621 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     4681 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     4741 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     4801 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     4861 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     4921 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4981 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     5041 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5101 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5161 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5221 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5281 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     5341 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5401 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5461 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5521 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     5581 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5641 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5701 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5761 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5821 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5881 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5941 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6001 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6061 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6121 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6181 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     6241 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6301 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6361 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6421 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6481 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6541 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6601 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6661 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6721 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6781 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6841 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6901 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6961 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7021 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7081 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7141 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7201 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7261 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7321 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     7381 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     7441 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     7501 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     7561 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7621 cttttgctgg ccttttgctc acatgt
//
