LOCUS       sequence-305748-        6373 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 6373)
  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 6373)
  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-Cre
FEATURES             Location/Qualifiers
     source          1..6373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   80..209
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        310..1488
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          541..1479
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1436..1456
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1507..1527
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    2579..3167
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2632..2652)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3050..3061)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3199..3675
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3398..3417)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3715..3855
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3715..3844
                     /label=AAV2 ITR
     rep_origin      3930..4385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4017..4036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4227..4248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4402..4421)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4521..4543
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4581..4599)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4667..4771
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4772..4840,4841..5632)
                     /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(4990..5009)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5803..6373,1..18)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6292..6311
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
       61 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
      121 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      181 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtaagcttt gcaaagatgg
      241 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
      301 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
      361 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
      421 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
      481 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
      541 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
      601 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
      661 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
      721 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
      781 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
      841 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
      901 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
      961 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
     1021 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     1081 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     1141 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     1201 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     1261 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     1321 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     1381 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     1441 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagg taccggatcc
     1501 accatgccca agaagaagag gaaggtgtcc aatctcctga ctgttcacca gaacctccct
     1561 gcgctgccag tagatgccac tagcgatgag gtcaggaaaa atctcatgga tatgtttagg
     1621 gatagacagg cgttttctga acacacctgg aaaatgctgc ttagcgtgtg ccgatcctgg
     1681 gcagcctggt gtaagctgaa caatcgcaaa tggttccccg ccgagccgga ggacgtgcgc
     1741 gattacctgc tgtatctcca ggcaagaggg ctggctgtca agactatcca gcagcacttg
     1801 ggccaactga atatgctgca tcgacgcagc gggctccccc ggcctagcga ttcaaacgca
     1861 gtctcccttg ttatgaggag aattagaaag gaaaacgtag atgcgggtga gagggctaag
     1921 caggctctcg cttttgagcg gactgatttc gaccaggtca gatccctgat ggagaacagc
     1981 gatcggtgcc aggacatcag gaacctcgca tttctgggaa ttgcatataa cacacttctg
     2041 cgcatagctg agatcgcccg gatcagagtg aaagacatca gtcgaacgga cggcggccgg
     2101 atgcttattc atattggacg cacaaagaca ttggtcagca ccgctggcgt tgaaaaggcc
     2161 ttgtccctgg gcgtaacgaa gctggtggaa agatggatct cagtgtccgg cgtggctgac
     2221 gaccctaata attacttgtt ctgtcgagtg agaaaaaacg gagtcgccgc gccctctgcc
     2281 accagccaat tgagtacacg ggcccttgaa gggatctttg aggcaaccca ccgactcata
     2341 tacggagcca aggatgacag tggccagagg tatctcgcct ggtcaggtca ttctgctagg
     2401 gtgggggccg cacgagacat ggcgcgggca ggagtctcca taccagagat tatgcaagct
     2461 ggaggttgga caaatgtgaa catcgttatg aactatatcc gcaatcttga ctctgaaacc
     2521 ggggccatgg tgagactgct cgaagatggt gactgaattc gatatcaagc ttatcgataa
     2581 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2641 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2701 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2761 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2821 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2881 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2941 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     3001 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     3061 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     3121 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc
     3181 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     3241 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     3301 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     3361 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     3421 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     3481 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     3541 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     3601 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     3661 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac
     3721 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3781 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3841 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3901 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     3961 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     4021 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4081 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4141 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4201 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4261 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     4321 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4381 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4441 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4501 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4561 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4621 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4681 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4741 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4801 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4861 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4921 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4981 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5041 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5101 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5161 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5221 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5281 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5341 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5401 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5461 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5521 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5581 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5641 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5701 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5761 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5821 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5881 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5941 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6001 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6061 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6121 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6181 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6241 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6301 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6361 atgctcgtca ggg
//
