LOCUS       sequence-218252-        7658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7658)
  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 7658)
  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-Cre
FEATURES             Location/Qualifiers
     source          1..7658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    132..608
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     390..409
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    640..1228
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             746..757
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1155..1175
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2280..2300)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2315..2867
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2477..2496)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2706..2723
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2884..3591)
                     /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(3162..3181)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3431..3449
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(3604..4782)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3613..4551
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3636..3656)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   4883..5012
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(5074..5662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5154..5173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5833..6624,6625..6693))
                     /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     6456..6475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6694..6798)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6866..6884
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6922..6944)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7044..7063
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7080..7535)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7217..7238)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7429..7448
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(7610..7658,1..92)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   join(7621..7658,1..92)
                     /label=AAV2 ITR
ORIGIN
        1 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
       61 gggagtggcc aactccatca ctaggggttc ctgcggccgc tcggtccgca cgtggttacc
      121 tacaaaatca gaaggacagg gaagggagca gtggttcacg cctgtaatcc cagcaatttg
      181 ggaggccaag gtgggtagat cacctgagat taggagttgg agaccagcct ggccaatatg
      241 gtgaaacccc gtctctacca aaaaaacaaa aattagctga gcctggtcat gcatgcctgg
      301 aatcccaaca actcgggagg ctgaggcagg agaatcgctt gaacccagga ggcggagatt
      361 gcagtgagcc aagattgtgc cactgcactc cagcttggtt cccaatagac cccgcaggcc
      421 ctacaggttg tcttcccaac ttgccccttg ctccatacca cccccctcca ccccataata
      481 ttatagaagg acacctagtc agacaaaatg atgcaactta attttattag gacaaggctg
      541 gtgggcactg gagtggcaac ttccagggcc aggagaggca ctggggaggg gtcacaggga
      601 tgccacccgt agatctctcg agcagcgctc ggtatcgatg cggggaggcg gcccaaaggg
      661 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
      721 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
      781 cgcgcagaat ccaggtggca acataggcga gcagccaagg aaaggacgat gatttccccg
      841 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
      901 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
      961 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     1021 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     1081 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     1141 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     1201 cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat tcagtcacca
     1261 tcttcgagca gtctcaccat ggccccggtt tcagagtcaa gattgcggat atagttcata
     1321 acgatgttca catttgtcca acctccagct tgcataatct ctggtatgga gactcctgcc
     1381 cgcgccatgt ctcgtgcggc ccccacccta gcagaatgac ctgaccaggc gagatacctc
     1441 tggccactgt catccttggc tccgtatatg agtcggtggg ttgcctcaaa gatcccttca
     1501 agggcccgtg tactcaattg gctggtggca gagggcgcgg cgactccgtt ttttctcact
     1561 cgacagaaca agtaattatt agggtcgtca gccacgccgg acactgagat ccatctttcc
     1621 accagcttcg ttacgcccag ggacaaggcc ttttcaacgc cagcggtgct gaccaatgtc
     1681 tttgtgcgtc caatatgaat aagcatccgg ccgccgtccg ttcgactgat gtctttcact
     1741 ctgatccggg cgatctcagc tatgcgcaga agtgtgttat atgcaattcc cagaaatgcg
     1801 aggttcctga tgtcctggca ccgatcgctg ttctccatca gggatctgac ctggtcgaaa
     1861 tcagtccgct caaaagcgag agcctgctta gccctctcac ccgcatctac gttttccttt
     1921 ctaattctcc tcataacaag ggagactgcg tttgaatcgc taggccgggg gagcccgctg
     1981 cgtcgatgca gcatattcag ttggcccaag tgctgctgga tagtcttgac agccagccct
     2041 cttgcctgga gatacagcag gtaatcgcgc acgtcctccg gctcggcggg gaaccatttg
     2101 cgattgttca gcttacacca ggctgcccag gatcggcaca cgctaagcag cattttccag
     2161 gtgtgttcag aaaacgcctg tctatcccta aacatatcca tgagattttt cctgacctca
     2221 tcgctagtgg catctactgg cagcgcaggg aggttctggt gaacagtcag gagattggac
     2281 accttcctct tcttcttggg catggttgtg gccatattat catcgtgttt ttcaaaggaa
     2341 aaccacgtcc ccgtggttcg gggggcctag acgttttttt aacctcgact aaacacatgt
     2401 aaagcatgtg caccgaggcc ccagatcaga tcccatacaa tggggtacct tctgggcatc
     2461 cttcagcccc ttgttgaata cgcttgagga gagccatttg actctttcca caactatcca
     2521 actcacaacg tggcactggg gttgtgccgc ctttgcaggt gtatcttata cacgtggctt
     2581 ttggccgcag aggcacctgt cgccaggtgg ggggttccgc tgcctgcaaa gggtcgctac
     2641 agacgttgtt tgtcttcaag aagcttccag aggaactgct tccttcacga cattcaacag
     2701 accttgcatt cctttggcga gaggggaaag acccctagga atgctcgtca agaagacagg
     2761 gccaggtttc cgggccctca cattgccaaa agacggcaat atggtggaaa ataacatata
     2821 gacaaacgca caccggcctt attccaagcg gcttcggcca gtaacgttag gggggggcgc
     2881 gccttacttg tacagctcgt ccatgccgcc ggtggagtgg cggccctcgg cgcgttcgta
     2941 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccaacttga tgttgacgtt
     3001 gtaggcgccg ggcagctgca cgggcttctt ggccttgtag gtggtcttga cctcagcgtc
     3061 gtagtggccg ccgtccttca gcttcagcct ctgcttgatc tcgcccttca gggcgccgtc
     3121 ctcggggtac atccgctcgg aggaggcctc ccagcccatg gtcttcttct gcattacggg
     3181 gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg
     3241 cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc
     3301 ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt
     3361 cacgtaggcc ttggagccgt acatgaactg aggggacagg atgtcccagg cgaagggcag
     3421 ggggccaccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc
     3481 gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgt gcaccttgaa
     3541 gcgcatgaac tccttgatga tggccatgtt atcctcctcg cccttgctca cggatccggt
     3601 acctcacgac acctgaaatg gaagaaaaaa actttgaacc actgtctgag gcttgagaat
     3661 gaaccaagat ccaaactcaa aaagggcaaa ttccaaggag aattacatca agtgccaagc
     3721 tggcctaact tcagtctcca cccactcagt gtggggaaac tccatcgcat aaaacccctc
     3781 cccccaacct aaagacgacg tactccaaaa gctcgagaac taatcgaggt gcctggacgg
     3841 cgcccggtac tccgtggagt cacatgaagc gacggctgag gacggaaagg cccttttcct
     3901 ttgtgtgggt gactcacccg cccgctctcc cgagcgccgc gtcctccatt ttgagctccc
     3961 tgcagcaggg ccgggaagcg gccatctttc cgctcacgca actggtgccg accgggccag
     4021 ccttgccgcc cagggcgggg cgatacacgg cggcgcgagg ccaggcacca gagcaggccg
     4081 gccagcttga gactaccccc gtccgattct cggtggccgc gctcgcaggc cccgcctcgc
     4141 cgaacatgtg cgctgggacg cacgggcccc gtcgccgccc gcggccccaa aaaccgaaat
     4201 accagtgtgc agatcttggc ccgcatttac aagactatct tgccagaaaa aaagcgtcgc
     4261 agcaggtcat caaaaatttt aaatggctag agacttatcg aaagcagcga gacaggcgcg
     4321 aaggtgccac cagattcgca cgcggcggcc ccagcgccca ggccaggcct caactcaagc
     4381 acgaggcgaa ggggctcctt aagcgcaagg cctcgaactc tcccacccac ttccaacccg
     4441 aagctcggga tcaagaatca cgtactgcag ccaggtggaa gtaattcaag gcacgcaagg
     4501 gccataaccc gtaaagaggc caggcccgcg ggaaccacac acggcactta cctgtgttct
     4561 ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc
     4621 tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc
     4681 gcgccacctt ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac
     4741 tgtgggcgat gtgcgctctg cccactgacg ggcaccggag ccaattccca ctcctttcaa
     4801 gacctagaag gtccattagc tgcaaagatt cctctctgtt taaaacttta tccatctttg
     4861 caaagcttac gcgtgcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     4921 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc
     4981 agtgagcgag cgagcgcgca gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc
     5041 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5101 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5161 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5221 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5281 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5341 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5401 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5461 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5521 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5581 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5641 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5701 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5941 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6781 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6841 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6901 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6961 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     7021 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
     7081 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     7141 ttttaaccaa tagaccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat
     7201 agagttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     7261 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa
     7321 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     7381 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     7441 gaaaggagcg ggcgctaagg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     7501 acccgccgcg cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg
     7561 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag
     7621 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaa
//
