LOCUS       sequence-334282-        6788 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Conditional expression of the eIF2alpha phophatase, CReP (Ppp1r15b).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6788)
  AUTHORS   Helseth AR, Hernandez-Martinez R, Hall VL, Oliver ML, Turner BD, 
            Caffall ZF, Rittiner JE, Shipman MK, King CS, Gradinaru V, Gerfen C,
            Costa-Mattioli M, Calakos N
  TITLE     Cholinergic neurons constitutively engage the ISR for dopamine 
            modulation and skill learning in mice.
  JOURNAL   Science. 2021 Apr 23;372(6540). pii: 372/6540/eabe1931. doi: 
            10.1126/science.abe1931.
  PUBMED    33888613
REFERENCE   2  (bases 1 to 6788)
  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: "Science."; 
            date: "2021-04-23"; volume: "372(6540). pii: 372/6540/eabe1931. 
            doi"; pages: " 10.1126/science.abe1931"
COMMENT     Sequence Label: AAV-hSyn-DIO-mCReP
FEATURES             Location/Qualifiers
     source          1..6788
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        233..680
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    696..729
                     /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    792..825
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2927..2956)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(2970..3003)
                     /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(3074..3107)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3140..3728
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3193..3213)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3611..3622)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3769..3976
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4146..4275
                     /label=AAV2 ITR
     rep_origin      complement(4512..5100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4592..4611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5271..6062,6063..6131))
                     /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     5894..5913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6132..6236)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6262..6717)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6399..6420)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6611..6630
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcg taggttaatt aagtgtctag actgcagagg gccctgcgta tgagtgcaag
      241 tgggttttag gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc
      301 cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga
      361 ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc
      421 cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt
      481 cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg
      541 ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc
      601 tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg
      661 tgtcgtgcct gagagcgcag tcgagaagaa ttcccataac ttcgtataaa gtatcctata
      721 cgaagttata tcaaaatagg aagaccaatg cttcaccatc gacccgaatt gccaagcatc
      781 accatcgacc cataacttcg tataatgtat gctatacgaa gttatactag ctagctcact
      841 tcttaacatt gctgtacaac agtaagtcct ttgactcgat cctcagtcta ttaaacattt
      901 tttctctgtg ctcaaaggcc aagcagtagc caatggcaac ttctgtttct tgaattcgtt
      961 tctggaacct gcatccatcc cttgcaaatt cttcccatgg tcctttgcga tcctcatcac
     1021 cacttatata atactcagta acttcttcca ggaaggttac cttttttctc ttgatatggg
     1081 tgtatctttc tccagcaaga gcctcaccca gcccacagcc tggacaatta tcttcttggc
     1141 tctctaacag ctgggcctta caagaaagta aggtatgatg gccagagaag gccactgtag
     1201 cctcagaaga ggcctttgag tcctgacggc ctttccaatt cttccctgac ggttgaaaag
     1261 gagccttaaa atttaaaagg ttgtaggggt cctcagaatg acagaaagag ttccacaatt
     1321 taagattctc tgcttcatct gcactcggtt cccagtcatc ttcctccccg gaactatggt
     1381 cgggggtctc cggaaggggt ccctcctgac agctccctac accacagctg ccagaccagg
     1441 atgtccctga atctgatggg tctctggggg caattctggc agccgtctga atcgtggctg
     1501 taaagttttg ggggttgtaa ggatctacac tgtggaaaga gttccaaagg tgaaggcctt
     1561 ccgagtcctg ttccacgtct gattcagaca gggagccatc gctatcaaag ccatcgtcct
     1621 caggttcctc gccccaatcc tcactttcag aatcagagct ggcttccaag tcactggggg
     1681 cgcctcccaa aatataatct atcagtttgt tgctacaaac tggtctggta gaaacaggaa
     1741 ggctctctcc tctatctgag aggtcagttg cattagtgtg gtcctcttca cattcttttt
     1801 ccacgggtaa ttcacaaaac agactgcttc cctgggggct ctgtgaaact tcttgaatca
     1861 ccaattctgg ttttttctca gtgggctccg gcctgtctgc agcaggggac accgcctgtg
     1921 ctgggttatc tgtgcaatgt tgtgggtcca tccggagaag gttatgttcc tcctccaggc
     1981 tatgatagcc attatcttgg tcaggggtgg gtaactcttg ccctttgtta gcctggagga
     2041 attctagccg tttcatacga aggtggtgga tttccggcag gccttctgta gagagaggcg
     2101 gacaccttct ccaggcggtt gccgacatct ctgcccgcaa cggctgaggg tgacaaccat
     2161 cttcagaaag atagcagctc tctggggtta gtgttcggaa acccacaaac tggccacctc
     2221 cagcgctgct ctgacagcgc agccctaact ggggaaggta gtccaggtag gaagggttca
     2281 ggagatagga aactaccttg aaattaccta agctctgaac gctcagaggc ccagaggatg
     2341 aagagtcaag ttctcggtgg gagactagac cagcttgaag gctactgggc agcaactcca
     2401 ctccccacag ctgctgctcc aggaggaaag tgggcgctgc agagtctaaa gctctttcct
     2461 gggccttgag ttttaactcc aggtctgagg acgagcactg ccactggagt ccctcctcta
     2521 gccaatcaag agaactgaca accaagtctt ccgaagagtc cagccgcagt gaactcagag
     2581 acttctgcac cgtgggagcg gcagattcct cccgtcctct caaagctctc agggcgctgt
     2641 aacttgcggc aaaatctagc catctggtag gaatcaggcc cccgaaaagc tggctccaaa
     2701 gcagcagctt ctggaatagg ctagggagca gggcaaggag ctgagaaagc aatttggtcc
     2761 agtacctggt ccgacgctcg ggcagagcgg agttcccggg attttgtcga gaggaaggcg
     2821 gcgggaactc cgaagagcag gcgtgcgatc gccgaaggaa gggcagccgg aaccaggagc
     2881 ccagccgagg gccaggccgc ttccgggccc tgtgcgttcc tgtctccaga tcctcttctg
     2941 agatgagttt ttgttccatg gtacctagta taacttcgta taggatactt tatacgaagt
     3001 tatcattggg attcttccta ttttgatcca agcatcacca tcgaccctct agtccagatc
     3061 tcaccatcga cccataactt cgtatagcat acattatacg aagttatgtc cctcgaagag
     3121 gttcgcgaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3181 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3241 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3301 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3361 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3421 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3481 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3541 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3601 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3661 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3721 ctccccgcat cgataccgtc gacccgggcg gccgctcata aagctcgact gtgccttcta
     3781 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     3841 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     3901 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     3961 gcaggcatgc tggggatgcg gtccggactg tacaattgca ttcattttat gtttcaggtt
     4021 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     4081 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     4141 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4201 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4261 cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4321 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4381 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4441 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4501 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4561 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4621 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4681 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4741 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4801 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4861 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4921 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4981 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5041 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5101 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5161 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5221 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5281 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5341 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5401 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5461 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5521 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5581 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5641 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5701 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5761 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5821 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5881 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5941 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6001 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6061 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6121 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6181 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6241 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     6301 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     6361 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     6421 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     6481 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     6541 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     6601 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     6661 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     6721 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     6781 tacgccag
//
