LOCUS       sequence-355958-        7475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses iCre linked to mCherry. The promoter is a fragment of the 
            hdc gene promoter which drives pan neuronal expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7475)
  AUTHORS   Miracca G, Anuncibay-Soto B, Tossell K, Yustos R, Vyssotski AL, 
            Franks NP, Wisden W
  TITLE     NMDA Receptors in the Lateral Preoptic Hypothalamus Are Essential 
            for Sustaining NREM and REM Sleep.
  JOURNAL   J Neurosci. 2022 May 31. pii: JNEUROSCI.0350-21.2022. doi: 
            10.1523/JNEUROSCI.0350-21.2022.
  PUBMED    35649726
REFERENCE   2  (bases 1 to 7475)
  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: "J Neurosci.
            2022 May 31. pii: JNEUROSCI.0350-21.2022. doi: 
            10.1523/JNEUROSCI.0350-21.2022."
COMMENT     Sequence Label: pAAV-iCre-2A-cherry
FEATURES             Location/Qualifiers
     source          1..7475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     CDS             1423..1443
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2458..2511
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2527..3237
                     /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(2672..2690)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2940..2959
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3301..3889
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3354..3374)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3772..3783)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3916..3933)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3922..4146
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4356..4685)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4370..4505
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4670..4690
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4931..5519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5011..5030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5690..6481,6482..6550))
                     /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     6313..6332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6551..6655)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6723..6741
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6779..6801)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6901..6920
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7134..7153
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7443..7461
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7443..7460
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctaggt accgaccacc ccccatgaag tctgttgtgt aagtacaggt aatgctttag
      241 acactgtttt tctcagattg tgttcaagct cttgggtgca ggcatccgca gtccagagcc
      301 tctgaggaac aaagggacct cgatataaag aacaaaaagc tctcggtatt cgggaaaact
      361 ccgaaggatt tttgttgttt tgtggcagtg ttggggttca ggacaaacac accaccccag
      421 aatatgattt caggagaaac tgccatcccc gcccccacta gtacgcttcc tttgtgttat
      481 ctcgagaagc cctggacgta ggaggagttc tcaaaagctg ttctttcgta agagatggtc
      541 aggaagaaac gttgattagg aaaagtactc gactcaggaa gaggaggggt acagataact
      601 gctgtcacca gaaagatttc catctgacaa agtggcaatt cttccccctt acgaatgtgt
      661 gtcccgcact ctgctgcacc caggctccgc tctgttctcc agctctgctt gctgtagaag
      721 cgtccatcca ctaaagggac ttccagattt attggcactg aacaggtttt tcctccgtta
      781 gcttaaacag tagtcgaggc aaagaaccca gaacaagtgc tatgtagtaa tggataaaaa
      841 gaatccttca tatcctgttg ttgatctgtc ctgagctcag cctggctagc tatgggcttc
      901 tctgagccgg ctctgccctc acctcataga gaagccaggg caggagcaac tccaccctct
      961 gtgctactta ttgtgaatgc ccttggccta tttctggcct ctgagctagg agctttattc
     1021 aaggctgggg ctctatggtc tgaatgcagg ggcagggatg atagagggct cttttacctt
     1081 gtctatccag agctgcttgt aggcttggga ggctaggtgc cccaactggt ctaaaaacac
     1141 catgttctct gattctgttg atcaaagttc aaatcagaaa ggacggggac aagagaggct
     1201 gaaattaagc ctgaaggaag ggactttatg ggggcggggc taaaggaggg gaatagggaa
     1261 ctccttaaat aagagatgca ctggctgcca gagagtgcac agcacagaca aaggcgacca
     1321 agaagccctg ttgacagctg ccttccagcc tcctctgtct gtctgccagg aggagcaatc
     1381 caaggaattc agagctcacg tcgaagcttg tccaccatgg tgcccaagaa gaagaggaaa
     1441 gtctccaacc tgctgactgt gcaccaaaac ctgcctgccc tccctgtgga tgccacctct
     1501 gatgaagtca ggaagaacct gatggacatg ttcagggaca ggcaggcctt ctctgaacac
     1561 acctggaaga tgctcctgtc tgtgtgcaga tcctgggctg cctggtgcaa gctgaacaac
     1621 aggaaatggt tccctgctga acctgaggat gtgagggact acctcctgta cctgcaagcc
     1681 agaggcctgg ctgtgaagac catccaacag cacctgggcc agctcaacat gctgcacagg
     1741 agatctggcc tgcctcgccc ttctgactcc aatgctgtgt ccctggtgat gaggagaatc
     1801 agaaaggaga atgtggatgc tggggagaga gccaagcagg ccctggcctt tgaacgcact
     1861 gactttgacc aagtcagatc cctgatggag aactctgaca gatgccagga catcaggaac
     1921 ctggccttcc tgggcattgc ctacaacacc ctgctgcgca ttgccgaaat tgccagaatc
     1981 agagtgaagg acatctcccg caccgatggt gggagaatgc tgatccacat tggcaggacc
     2041 aagaccctgg tgtccacagc tggtgtggag aaggccctgt ccctgggggt taccaagctg
     2101 gtggagagat ggatctctgt gtctggtgtg gctgatgacc ccaacaacta cctgttctgc
     2161 cgggtcagaa agaatggtgt ggctgcccct tctgccacct cccaactgtc cacccgggcc
     2221 ctggaaggga tctttgaggc cacccaccgc ctgatctatg gtgccaagga tgactctggg
     2281 cagagatacc tggcctggtc tggccactct gccagagtgg gtgctgccag ggacatggcc
     2341 agggctggtg tgtccatccc tgaaatcatg caggctggtg gctggaccaa tgtgaacatt
     2401 gtgatgaact acatcagaaa cctggactct gagactgggg ccatggtgag gctgctcgag
     2461 ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tgcaccggga
     2521 tccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     2581 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     2641 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     2701 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     2761 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     2821 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2881 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2941 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     3001 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     3061 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     3121 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     3181 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaggc
     3241 tagcagaatt ctgcagatat ccagcacagt ggcggccgca agctttaaca gcttatcgat
     3301 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3361 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3421 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3481 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3541 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3601 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3661 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3721 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3781 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3841 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     3901 cgactcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3961 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4021 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4081 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     4141 ctatggcttc tgaggcggaa agaaccagct ggggctcgac tagagcatgg ctacgtagat
     4201 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     4261 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4321 ggcggcctca gtgagcgagc gagcgcgcag agctttttgc aaaagcctag gcctccaaaa
     4381 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     4441 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
     4501 gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
     4561 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
     4621 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
     4681 cacacattcc acagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4741 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4801 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4861 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4921 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4981 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5041 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5101 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5161 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5221 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5281 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5341 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5401 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5461 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5521 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5581 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5641 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5701 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5761 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5821 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5881 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5941 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6001 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6061 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6121 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6181 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6241 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6301 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6361 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6421 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6481 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6541 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6601 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6661 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6721 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     6781 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     6841 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     6901 cggcatcaga gcagattgta ctgagagtgc accattcgac gctctccctt atgcgactcc
     6961 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     7021 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     7081 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     7141 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     7201 ccggcgtaga ggatctggct agcgatgacc ctgctgattg gttcgctgac catttccggg
     7261 tgcgggacgg cgttaccaga aactcagaag gttcgtccaa ccaaaccgac tctgacggca
     7321 gtttacgaga gagatgatag ggtctgcttc agtaagccag atgctacaca attaggcttg
     7381 tacatattgt cgttagaacg cggctacaat taatacataa ccttatgtat catacacata
     7441 cgatttaggt gacactatag aatacacgga attaa
//
