LOCUS       sequence-243346-        7636 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 7636)
  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 7636)
  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..7636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1422..2129
                     /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(1564..1582)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1832..1851
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2146..2698
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2290..2307)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2517..2536
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3763..4351
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3816..3836)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4234..4245)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4383..4859
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4582..4601)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4899..5039
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4899..5028
                     /label=AAV2 ITR
     rep_origin      5114..5569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5201..5220)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5411..5432
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5586..5605)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5705..5727
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5765..5783)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5851..5955
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5956..6024,6025..6816)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGNQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6174..6193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6987..7575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7476..7495
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgtgagcaag ggcgaggagg
     1441 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     1501 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1561 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     1621 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     1681 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1741 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     1801 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     1861 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     1921 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     1981 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     2041 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     2101 ccggcggcat ggacgagctg tacaagtaag gcgcgccccc ccctaacgtt actggccgaa
     2161 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     2221 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     2281 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     2341 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     2401 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     2461 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     2521 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     2581 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     2641 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg
     2701 gccacaacca tgtcagagct tataatttca ggtagtagtg gaggatttct cagaaacatt
     2761 ggcaaggagt accaagaagc tgccgaaaat ttcatgaggt ttatgaacga ccagggcgca
     2821 tacgctccaa atactttgag agacctccga ctcgtgttcc acagctgggc tcgatggtgc
     2881 cacgccaggc agcttgcatg gtttccaata tctccagaaa tggctcgcga gtactttttg
     2941 caactccacg acgctgatct ggcatctacc accatcgaca agcactatgc aatgctgaat
     3001 atgctgctgt cccactgtgg actgcctcca ctgtcagatg acaagtccgt gagcctggcg
     3061 atgcggagaa ttagaagaga ggcggccact gaaaaggggg aacggacagg acaggcgata
     3121 cctctccgct gggacgacct gaaactgctg gacgtactcc tgagtcggag cgagcgcttg
     3181 gtcgatcttc ggaaccgcgc cttcctcttc gtggcctata ataccctgat gcggatgagt
     3241 gagatcagca gaatccgagt tggggatctg gatcagaccg gggacactgt gactttgcat
     3301 atctcccata ctaagactat caccaccgct gccggcttgg acaaggttct ctctaggcgc
     3361 acgaccgctg tcctgaatga ctggcttgat gtatctggcc tgcgagaaca tcccgacgcc
     3421 gtgctgttcc cccctattca ccggtctaac aaggcaagaa tcacgacaac gccgctgact
     3481 gcacccgcca tggagaagat cttttccgat gcttgggtgc tccttaacaa aagggatgca
     3541 acacctaaca aaggtagata caggacatgg accgggcata gcgcgcgggt gggggccgcg
     3601 attgatatgg ccgagaaaca ggtcagcatg gtggagatca tgcaggaggg tacatggaaa
     3661 aaaccggaga cactgatgag atatcttagg agggggggcg tcagcgttgg agcaaatagc
     3721 cgcctgatgg acagttgaga attcgatatc aagcttatcg ataatcaacc tctggattac
     3781 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3841 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3901 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3961 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     4021 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4081 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4141 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     4201 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4261 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4321 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct
     4381 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     4441 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     4501 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     4561 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     4621 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     4681 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     4741 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     4801 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     4861 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     4921 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4981 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     5041 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     5101 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5161 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     5221 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5281 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     5341 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5401 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     5461 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     5521 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     5581 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5641 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5701 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5761 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5821 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5881 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5941 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6001 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6061 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6121 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6181 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6241 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6301 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6361 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6421 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6481 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6541 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6601 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6661 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6721 atcgtagtta tctacacgac ggggaatcag gcaactatgg atgaacgaaa tagacagatc
     6781 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6841 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6901 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7441 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7501 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7561 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7621 ccttttgctc acatgt
//
