LOCUS       sequence-159339-        7121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cre-dependent PAK1-AID under EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7121)
  AUTHORS   Kong D, Dagon Y, Campbell JN, Guo Y, Yang Z, Yi X, Aryal P, 
            Wellenstein K, Kahn BB, Sabatini BL, Lowell BB
  TITLE     A Postsynaptic AMPK-->p21-Activated Kinase Pathway Drives 
            Fasting-Induced Synaptic Plasticity in AgRP Neurons.
  JOURNAL   Neuron. 2016 Jun 16. pii: S0896-6273(16)30204-5. doi: 
            10.1016/j.neuron.2016.05.025.
  PUBMED    27321921
REFERENCE   2  (bases 1 to 7121)
  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"; journalName: "Neuron. 
            2016 Jun 16. pii: S0896-6273(16)30204-5. doi: 
            10.1016/j.neuron.2016.05.025."
COMMENT     Sequence Label: pAAV-EF1a-Flex-PAK1-AID-ires-EGFP
FEATURES             Location/Qualifiers
     source          1..7121
                     /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"
     protein_bind    complement(1450..1483)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1534..1567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1576..2289,2290..2292,2293..2295))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1579..2289,2290..2292,2293..2295))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1621..1642)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1970..1989
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2229..2250
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    join(2296..2304,2305..2307,2308..2882)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2469..2488)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2698..2715
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    3106..3139
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3190..3223
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3248..3836
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3301..3321)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3719..3730)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3868..4344
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4067..4086)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4384..4524
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4384..4513
                     /label=AAV2 ITR
     rep_origin      4599..5054
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4686..4705)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4896..4917
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5071..5090)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5190..5212
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5250..5268)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5336..5440
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5441..5509,5510..6301)
                     /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(5659..5678)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6472..7060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6961..6980
                     /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 ctctagagtc
     1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     1561 aagttatggc gcgccttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     1621 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga
     1681 ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1741 ctgctggtag tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa
     1801 gttcaccttg atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta
     1861 gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag
     1921 ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt
     1981 gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt
     2041 gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccgtaggt
     2101 cagggtggtc acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt
     2161 cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg
     2221 gccgtttacg tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc
     2281 gcccttgctc accatggttg tggccatatt atcatcgtgt ttttcaaagg aaaaccacgt
     2341 ccccgtggtt cggggggcct agacgttttt ttaacctcga ctaaacacat gtaaagcatg
     2401 tgcaccgagg ccccagatca gatcccatac aatggggtac cttctgggca tccttcagcc
     2461 ccttgttgaa tacgcttgag gagagccatt tgactctttc cacaactatc caactcacaa
     2521 cgtggcactg gggttgtgcc gcctttgcag gtgtatctta tacacgtggc ttttggccgc
     2581 agaggcacct gtcgccaggt ggggggttcc gctgcctgca aagggtcgct acagacgttg
     2641 tttgtcttca agaagcttcc agaggaactg cttccttcac gacattcaac agaccttgca
     2701 ttcctttggc gagaggggaa agacccctag gaatgctcgt caagaagaca gggccaggtt
     2761 tccgggccct cacattgcca aaagacggca atatggtgga aaataacata tagacaaacg
     2821 cacaccggcc ttattccaag cggcttcggc cagtaacgtt aggggggggg gagggagagg
     2881 ggcctcgagt tatgacttat ctgtaaaact catgtacttc tgactattgg aggtcttctt
     2941 agagttataa aattccaaca catccagaac agcctgtggg tttttcttct gttctgactt
     3001 tgtgatattt gatgtttgaa gcaagcgagc ccactgttct ggcattcccg taaactcccc
     3061 tgtgacagca tcaaaaccaa catgaattgt atgcattgtg ctagcataac ttcgtataaa
     3121 gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg
     3181 gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg atatcaagct
     3241 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3301 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3361 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3421 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3481 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3541 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3601 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3661 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3721 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3781 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3841 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     3901 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     3961 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     4021 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     4081 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     4141 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     4201 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     4261 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     4321 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     4381 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4441 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4501 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     4561 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     4621 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4681 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4741 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4801 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4861 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4921 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     4981 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5041 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5101 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5161 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5221 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     5281 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5341 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5401 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5461 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5521 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5581 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5641 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5701 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5761 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5821 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5881 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5941 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6001 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6061 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6121 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6181 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6241 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6301 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6361 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6421 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6481 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6541 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6601 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6661 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6721 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6781 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6841 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6901 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6961 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7021 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7081 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
