LOCUS       sequence-86308-e        6451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sCre in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6451)
  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 6451)
  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-sCre
FEATURES             Location/Qualifiers
     source          1..6451
                     /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"
     misc_feature    2578..3166
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2631..2651)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3049..3060)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3198..3674
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3397..3416)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3714..3854
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3714..3843
                     /label=AAV2 ITR
     rep_origin      3929..4384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4016..4035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4226..4247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4401..4420)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4520..4542
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4580..4598)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4666..4770
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4771..4839,4840..5631)
                     /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(4989..5008)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5802..6390
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6291..6310
                     /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 cgccaccatg
     1441 agcctgctga ctaccaacaa ccattctgtg gcactgtcct acggcgaacc tccctctact
     1501 ctgaacgatt ctctgaaaga ttcctaccag aggtctaccg acgagctgca ggcactgctg
     1561 agtaagccac tggcacagct gaccgacgcc gataaactgc ggatcagaga aattacacag
     1621 gctaagctga aacactttct ggataacggc catagaactc ggagagctaa tacctggaga
     1681 gcactgatga gtaggtgggc caagttcgag tcatggtgcc tgaccaacaa tctgacaccc
     1741 ctgcctgcaa cacccgaagt ggtcgccact tttatcgagt actatcaggc cagctcctac
     1801 accacactga gccagtatgc ttgggcaatt aactccttcc acgtggaatg tggactgctg
     1861 tctcccgtgt ctagtaagac agtccaggac aaacagaatg aaatccggat tgtgaagctg
     1921 gagagcggag ggctggccca ggaacaggct actcctttta gactgcacca tctgcagatg
     1981 ctgatcgagt cttacgggga aagtgagcgc ctgctggata aacgaaacct ggccctgctg
     2041 aatatcgctt atgagtccct gctgcgggaa tctgagctgc tgagaattaa ggtcggccat
     2101 ctgaaatcaa cattcgaagg ggactacgtg ctgagcgtcc cttatactaa gaccaacgat
     2161 tccggggagg aagaggtggt caatatcact ccactgggat tcaagctgat ccagaggtac
     2221 attcagggag ccggcctgac caaagaggac tatctgtttc agccaattgg ccgctcaaac
     2281 aaggtgagcg tccaggccaa acccatgagc acacgaactg tggaccgggt cttcctgtgg
     2341 gcttttgaat ccctggggat cgatcgacac tcagcttgga gcgggcattc cgcacgcatt
     2401 ggagccgctc aggatctgct ggcagccgga tattctatcg cccagattca ggagaatggc
     2461 aggtggaaaa gtcctatgat ggtgctgcgc tacggcaaag acatcaaggc aaaagaatcc
     2521 gctatggcaa aaatgctggc agaaagaaga taagaattcg atatcaagct tatcgataat
     2581 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2641 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2701 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2761 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     2821 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     2881 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     2941 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3001 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3061 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3121 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg
     3181 ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     3241 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     3301 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     3361 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     3421 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     3481 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     3541 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     3601 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     3661 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc
     3721 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3781 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3841 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3901 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     3961 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4021 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4081 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4141 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4201 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4261 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4321 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4381 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4441 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4501 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4561 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4621 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4681 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4741 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4801 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4861 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4921 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4981 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5041 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5101 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5161 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5221 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5281 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5341 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5401 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5461 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5521 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5581 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5641 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5701 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5761 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5821 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5881 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5941 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6001 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6061 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6121 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6181 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6241 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6301 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6361 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6421 ctggcctttt gctggccttt tgctcacatg t
//
