LOCUS       sequence-198690-        7405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of hM4Di-mCherry under CBA promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7405)
  AUTHORS   Hou XH, Hyun M, Taranda J, Huang KW, Todd E, Feng D, Atwater E, 
            Croney D, Zeidel ML, Osten P, Sabatini BL
  TITLE     Central Control Circuit for Context-Dependent Micturition.
  JOURNAL   Cell. 2016 Sep 22;167(1):73-86.e12. doi: 10.1016/j.cell.2016.08.073.
  PUBMED    27662084
REFERENCE   2  (bases 1 to 7405)
  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.2016.08"; journalName: "Cell"; date: "2016-09-22- 
            22"; volume: "167"; issue: "1"; pages: "73-86.e12"
COMMENT     Sequence Label: pAAV-CBA-DIO-hM4Di-mCherry
FEATURES             Location/Qualifiers
     source          1..7405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     1573..1595
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1657..1676
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(1733..1766)
                     /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(1817..1850)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1859..2569)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2137..2156)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2406..2424
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2566..2575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4024..4033
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4040..4073
                     /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    4124..4157
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4182..4770
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4235..4255)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4653..4664)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4802..5278
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5001..5020)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5318..5458
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5318..5447
                     /label=AAV2 ITR
     rep_origin      5533..5988
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5620..5639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5830..5851
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6005..6024)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6124..6146
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6184..6202)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6270..6374
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6375..6443,6444..7235)
                     /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(6593..6612)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      841 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      901 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      961 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     1021 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     1081 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     1141 cgatgggggc gggggggggg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     1201 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     1261 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgacgct gccttcgccc
     1321 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
     1381 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
     1441 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
     1501 tttgtgcggg gggagcggct cggggctgtc cgcgggggga cggctgcctt cgggggggac
     1561 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
     1621 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     1681 ctcatcattt tggcaaagaa ttggtaccgg atcctctaga gtcgactccg gaataacttc
     1741 gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc tgctattagc
     1801 aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat ggcgcgcctt
     1861 acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt
     1921 ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg
     1981 cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt
     2041 ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg
     2101 ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt
     2161 cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg
     2221 aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact
     2281 tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt
     2341 aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc
     2401 cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct
     2461 cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca
     2521 tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatg gtggcgaccg
     2581 gtggatcccg cctggcagtg ccgatgttcc gatactggca cagcagcagg tgccggaagg
     2641 tctttttaaa ggtggcgttg cacagagcat agcaggcagg gttgatggtg ctgttgacgt
     2701 agcagagcca gtagccaatg gaccacaccg tgtcagggat gcagctctgg cagaaggtgt
     2761 tcaccaggac catgacgttg tagggcgtcc aggtgaggat gaaggccagc agaatggcaa
     2821 agatcgttcg tgtcactttg cgctcccggg ccgccatctg ccgcttcttg cgcacctggt
     2881 tgcgagcgat gctggcgaac ttgcgggcca cgttggccgc agggcgcatg ccagccggcg
     2941 tggcaggcac aatctcaatg gctgtcacac actcattgcc tgtctgcttc gtcacaatct
     3001 ggatcttgga ccatctggag gctgggttga gggcccgcgg ctgcagggga ggggcgggca
     3061 tggcgggcgt ggtggcctct gtggtggaca gctctgtggc tgggcgttcc ttggtgttct
     3121 gggtggcact gcctgagctg gactcattgg aagtgtcctt atcagccacg gggcgcggtg
     3181 gcggtggcag cgctggcggg ggggcctcct ccagcttgcc attgcgcagc tcctcccggg
     3241 cggcctcccc gggcgggggc ttcttgacgc tctgcttcat tagtgggctc ttgaggaagg
     3301 ccagcgtctt ggctttcttc tccttcgggc cctcgggccg gtgcttgtgg actcggctgc
     3361 gactggccag ggagatgtgg atgtacagca ccgtcatgat gaccacaggc aggtagaagc
     3421 cagcaatggc tgtgccaaag gtcactgctg ggttggacag gaactggatg aagcactggt
     3481 tgtcgggcac cgtccgctta cccaccacaa actgccagaa caagatggca ggcgcccaga
     3541 gcacgaagga cagtacccag gcagcagcaa tcatgaggcc tgccatcttg gtggtgcgcc
     3601 gggcagggta ggtgagaggc ttggtgacgc agaagtagcg gtcaaagctg atgatgagaa
     3661 ggttcatgac ggaggcgttg ctcaccacgc agtccagggc cagccacagg tcgcagacca
     3721 cggcgcccag gggccagtag cccttgatga tgtacacggt gtagaggttc atggagaagg
     3781 cgcctatgat gagatcagca cacgccaggc tgaagaggaa gtagttgttg actgtctgca
     3841 gctgcctgtt gaccttgatg gacagcatca ccaggatgtt gcccacgaca gtcaccaggc
     3901 tcagggagcc tgtcactgtg gcaatgaaga ccatttccac cgtctcatag cgattgtggg
     3961 atgatgacgt gaccaggcgc acggactgat tgcccgagct gccattgaca ggtgtgaagt
     4021 tggccatggt ggcgctagca taacttcgta taaagtatcc tatacgaagt tatttgcctt
     4081 aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt
     4141 atgctatacg aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca
     4201 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     4261 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     4321 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     4381 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     4441 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     4501 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     4561 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     4621 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     4681 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     4741 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     4801 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     4861 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     4921 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     4981 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     5041 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     5101 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     5161 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     5221 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     5281 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     5341 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     5401 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     5461 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5521 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5581 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     5641 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5701 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5761 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5821 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     5881 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     5941 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     6001 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     6061 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6121 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6181 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6241 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6301 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6361 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6421 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6481 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6541 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6601 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6661 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6721 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6781 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6841 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6901 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6961 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7021 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7081 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7141 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7201 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7261 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7321 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7381 ccgtagaaaa gatcaaagga tcttc
//
