LOCUS       sequence-434470-        6475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry protein in infected and cre positive cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6475)
  AUTHORS   de Leon Reyes NS, Sierra Diaz P, Nogueira R, Ruiz-Pino A, Nomura Y, 
            de Solis CA, Schulkin J, Asok A, Leroy F
  TITLE     Corticotropin-releasing hormone signaling from prefrontal cortex to 
            lateral septum suppresses interaction with familiar mice.
  JOURNAL   Cell. 2023 Sep 14;186(19):4152-4171.e31. doi: 
            10.1016/j.cell.2023.08.010. Epub 2023 Sep 4.
  PUBMED    37669667
REFERENCE   2  (bases 1 to 6475)
  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: 
            "10.1016/j.cell.2023.08.010"; journalName: "Cell"; date: 
            "2023-09-14- 14"; volume: "186"; issue: "19"; pages: "4152-4171.e31"
COMMENT     Sequence Label: pAAV CMV-DIO-(mCherry-U6)-shRNA(scrambled)
FEATURES             Location/Qualifiers
     source          1..6475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     710..734
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          862..1240
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     protein_bind    complement(1317..1350)
                     /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(1401..1434)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        complement(1439..1679)
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     primer_bind     complement(1489..1508)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1659..1679)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(1686..2396)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEYNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1964..1983)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2233..2251
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2393..2402
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2409..2442
                     /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    2493..2526
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2602..3190
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2655..2675)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3073..3084)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3222..3698
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3421..3440)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3738..3878
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3738..3867
                     /label=AAV2 ITR
     rep_origin      3953..4408
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4040..4059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4250..4271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4425..4444)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4544..4566
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4604..4622)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4690..4794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4795..4863,4864..5655)
                     /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(5013..5032)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5826..6414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6315..6334
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      721 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      781 gggaccgatc cagcctccgc ggattcgaat cccggccggg aacggtgcat tggaacgcgg
      841 attccccgtg ccaagagtga cgtaagtacc gcctatagag tctataggcc cacaaaaaat
      901 gctttcttct tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct
      961 ttctttcagg gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa
     1021 cagtgataat ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata
     1081 taaattgtaa ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca
     1141 ttctgctttt attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct
     1201 tttgctaatc atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt
     1261 ctgtgtgctg gcccatcact ttggcaaaga attgggattc tagagtcgac tccggaataa
     1321 cttcgtatag gatactttat acgaagttat gcagaatggt agctggattg tagctgctat
     1381 tagcaatatg aaacctctta ataacttcgt atagcataca ttatacgaag ttattttcgt
     1441 cctttccaca agatatataa agccaagaaa tcgaaatact ttcaagttac ggtaagcata
     1501 tgatagtcca ttttaaaaca taattttaaa actgcaaact acccaagaaa ttattacttt
     1561 ctacgtcacg tattttgtac taatatcttt gtgtttacag tcaaattaat tccaattatc
     1621 tctctaacag ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa taggccctcg
     1681 gatccttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
     1741 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
     1801 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
     1861 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     1921 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     1981 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     2041 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     2101 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     2161 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     2221 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     2281 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     2341 aagcgcatga actccttgat gatggccatg ttatactcct cgcccttgct caccatggtg
     2401 gcgctagcat aacttcgtat aaagtatcct atacgaagtt atttgcctta acccagaaat
     2461 tatcactgtt attctttaga atggtgcaaa gaataacttc gtataatgta tgctatacga
     2521 agttatcacc ggcctaaggt taagtcgccc tcgttcaaga gacgagggcg acttaacctt
     2581 aggctttttt aattcactag taatcaacct ctggattaca aaatttgtga aagattgact
     2641 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2701 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2761 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2821 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2881 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2941 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3001 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3061 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3121 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3181 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     3241 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     3301 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     3361 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     3421 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     3481 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     3541 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     3601 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     3661 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     3721 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     3781 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     3841 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     3901 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     3961 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4021 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4081 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4141 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     4201 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4261 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     4321 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4381 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     4441 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4501 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4561 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     4621 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4681 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4741 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4921 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4981 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5041 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5101 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5161 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5221 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5281 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5341 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5401 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5461 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5521 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5581 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5641 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5701 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5761 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5821 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5881 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5941 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6001 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6061 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6121 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6181 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6241 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6301 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6361 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6421 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
