LOCUS       sequence-325404-        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"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2691..3138
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    3154..3187
                     /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    3250..3283
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(5385..5414)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(5428..5461)
                     /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(5532..5565)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5598..6186
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5651..5671)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6069..6080)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6227..6434
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6604..6733
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg
     2701 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca
     2761 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg
     2821 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct
     2881 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca
     2941 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc
     3001 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg
     3061 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg
     3121 tcgtgcctga gagcgcagtc gagaagaatt cccataactt cgtataaagt atcctatacg
     3181 aagttatatc aaaataggaa gaccaatgct tcaccatcga cccgaattgc caagcatcac
     3241 catcgaccca taacttcgta taatgtatgc tatacgaagt tatactagct agctcacttc
     3301 ttaacattgc tgtacaacag taagtccttt gactcgatcc tcagtctatt aaacattttt
     3361 tctctgtgct caaaggccaa gcagtagcca atggcaactt ctgtttcttg aattcgtttc
     3421 tggaacctgc atccatccct tgcaaattct tcccatggtc ctttgcgatc ctcatcacca
     3481 cttatataat actcagtaac ttcttccagg aaggttacct tttttctctt gatatgggtg
     3541 tatctttctc cagcaagagc ctcacccagc ccacagcctg gacaattatc ttcttggctc
     3601 tctaacagct gggccttaca agaaagtaag gtatgatggc cagagaaggc cactgtagcc
     3661 tcagaagagg cctttgagtc ctgacggcct ttccaattct tccctgacgg ttgaaaagga
     3721 gccttaaaat ttaaaaggtt gtaggggtcc tcagaatgac agaaagagtt ccacaattta
     3781 agattctctg cttcatctgc actcggttcc cagtcatctt cctccccgga actatggtcg
     3841 ggggtctccg gaaggggtcc ctcctgacag ctccctacac cacagctgcc agaccaggat
     3901 gtccctgaat ctgatgggtc tctgggggca attctggcag ccgtctgaat cgtggctgta
     3961 aagttttggg ggttgtaagg atctacactg tggaaagagt tccaaaggtg aaggccttcc
     4021 gagtcctgtt ccacgtctga ttcagacagg gagccatcgc tatcaaagcc atcgtcctca
     4081 ggttcctcgc cccaatcctc actttcagaa tcagagctgg cttccaagtc actgggggcg
     4141 cctcccaaaa tataatctat cagtttgttg ctacaaactg gtctggtaga aacaggaagg
     4201 ctctctcctc tatctgagag gtcagttgca ttagtgtggt cctcttcaca ttctttttcc
     4261 acgggtaatt cacaaaacag actgcttccc tgggggctct gtgaaacttc ttgaatcacc
     4321 aattctggtt ttttctcagt gggctccggc ctgtctgcag caggggacac cgcctgtgct
     4381 gggttatctg tgcaatgttg tgggtccatc cggagaaggt tatgttcctc ctccaggcta
     4441 tgatagccat tatcttggtc aggggtgggt aactcttgcc ctttgttagc ctggaggaat
     4501 tctagccgtt tcatacgaag gtggtggatt tccggcaggc cttctgtaga gagaggcgga
     4561 caccttctcc aggcggttgc cgacatctct gcccgcaacg gctgagggtg acaaccatct
     4621 tcagaaagat agcagctctc tggggttagt gttcggaaac ccacaaactg gccacctcca
     4681 gcgctgctct gacagcgcag ccctaactgg ggaaggtagt ccaggtagga agggttcagg
     4741 agataggaaa ctaccttgaa attacctaag ctctgaacgc tcagaggccc agaggatgaa
     4801 gagtcaagtt ctcggtggga gactagacca gcttgaaggc tactgggcag caactccact
     4861 ccccacagct gctgctccag gaggaaagtg ggcgctgcag agtctaaagc tctttcctgg
     4921 gccttgagtt ttaactccag gtctgaggac gagcactgcc actggagtcc ctcctctagc
     4981 caatcaagag aactgacaac caagtcttcc gaagagtcca gccgcagtga actcagagac
     5041 ttctgcaccg tgggagcggc agattcctcc cgtcctctca aagctctcag ggcgctgtaa
     5101 cttgcggcaa aatctagcca tctggtagga atcaggcccc cgaaaagctg gctccaaagc
     5161 agcagcttct ggaataggct agggagcagg gcaaggagct gagaaagcaa tttggtccag
     5221 tacctggtcc gacgctcggg cagagcggag ttcccgggat tttgtcgaga ggaaggcggc
     5281 gggaactccg aagagcaggc gtgcgatcgc cgaaggaagg gcagccggaa ccaggagccc
     5341 agccgagggc caggccgctt ccgggccctg tgcgttcctg tctccagatc ctcttctgag
     5401 atgagttttt gttccatggt acctagtata acttcgtata ggatacttta tacgaagtta
     5461 tcattgggat tcttcctatt ttgatccaag catcaccatc gaccctctag tccagatctc
     5521 accatcgacc cataacttcg tatagcatac attatacgaa gttatgtccc tcgaagaggt
     5581 tcgcgaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     5641 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     5701 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     5761 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     5821 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     5881 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     5941 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     6001 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     6061 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     6121 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     6181 ccccgcatcg ataccgtcga cccgggcggc cgctcataaa gctcgactgt gccttctagt
     6241 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6301 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     6361 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     6421 aggcatgctg gggatgcggt ccggactgta caattgcatt cattttatgt ttcaggttca
     6481 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     6541 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     6601 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     6661 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     6721 agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6781 tgggcgct
//
