LOCUS       sequence-348532-        7473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DREADD hM4D (Gi) only in Cre expressing cells and not in 
            Flp expressing cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7473)
  TITLE     INTRSECT DREADD Gi plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7473)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-nEF-Con/Foff DREADD Gi-mCherry
FEATURES             Location/Qualifiers
     source          1..7473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        177..388
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             401..669
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    697..730
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     regulatory      823..832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1437..1470)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    complement(1521..1554)
                     /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)."
     primer_bind     1753..1771
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    2891..2924
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    2975..3008
                     /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)."
     primer_bind     3154..3173
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3458..3491)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    3600..4188
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3653..3673)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4071..4082)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4220..4696
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4419..4438)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4736..4876
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4736..4865
                     /label=AAV2 ITR
     rep_origin      4951..5406
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5038..5057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5248..5269
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5423..5442)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5542..5564
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5602..5620)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5688..5792
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5793..5861,5862..6653)
                     /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(6011..6030)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6824..7412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7313..7332
                     /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 gcgtaaggat ctgcgatcgc tccggtgccc gtcagtgggc
      181 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
      241 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
      301 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
      361 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
      421 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
      481 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
      541 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
      601 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
      661 ttacagatcc aagctgtgac cggcgcctac ggatccgaag ttcctattct tcaaatagta
      721 taggaacttc gcagagtagt agctggattg tagctgctat tagcaatcta aaacctctta
      781 gaagttccta ttcttcaaaa ggtataggaa cttcggcgcg ccgccaccat ggccaacttc
      841 acacctgtca atggcagctc gggcaatcag tccgtgcgcc tggtcacgtc atcatcccac
      901 aatcgctatg agacggtgga aatggtcttc attgccacag tgacaggctc cctgagcctg
      961 gtgactgtcg tgggcaacat cctggtgatg ctgtccatca aggtcaacag gcagctgcag
     1021 acagtcaaca actacttcct cttcagcctg gcgtgtgctg atctcatcat aggcgccttc
     1081 tccatgaacc tctacaccgt gtacatcatc aagggctact ggcccctggg cgccgtggtc
     1141 tgcgacctgt ggctggccct ggactgcgtg gtgagcaacg cctccgtcat gaaccttctc
     1201 atcatcagct ttgaccgcta cttctgcgtc accaagcctc tcacctaccc tgcccggcgc
     1261 accaccaaga tggcaggcct catgattgct gctgcctggg tactgtcctt cgtgctctgg
     1321 gcgcctgcca tcttgttctg gcagtttgtg gtgggtaagc ggacggtgcc cgacaaccag
     1381 tgcttcatcc agttcctgtc caacccagca gtgacgtaag tgtcggggtt gctagcataa
     1441 cttcgtatag tataccttat acgaagttat tctttgcacc attctaaaga ataacagtga
     1501 taatttctgg gttaaggcaa ataacttcgt ataggatact ttatacgaag ttatggtacc
     1561 cctcctgtac tcacctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     1621 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     1681 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     1741 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     1801 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     1861 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct caccatggta
     1921 gcgaccggtg gatcccgcct ggcagtgccg atgttccgat actggcacag cagcaggtgc
     1981 cggaaggtct ttttaaaggt ggcgttgcac agagcatagc aggcagggtt gatggtgctg
     2041 ttgacgtagc agagccagta gccaatggac cacaccgtgt cagggatgca gctctggcag
     2101 aaggtgttca ccaggaccat gacgttgtag ggcgtccagg tgaggatgaa ggccagcaga
     2161 atggcaaaga tcgttcgtgt cactttgcgc tcccgggccg ccatctgccg cttcttgcgc
     2221 acctggttgc gagcgatgct ggcgaacttg cgggccacgt tggccgcagg gcgcatgcca
     2281 gccggcgtgg caggcacaat ctcaatggct gtcacacact cattgcctgt ctgcttcgtc
     2341 acaatctgga tcttggacca tctggaggct gggttgaggg cccgcggctg caggggaggg
     2401 gcgggcatgg cgggcgtggt ggcctctgtg gtggacagct ctgtggctgg gcgttccttg
     2461 gtgttctggg tggcactgcc tgagctggac tcattggaag tgtccttatc agccacgggg
     2521 cgcggtggcg gtggcagcgc tggcgggggg gcctcctcca gcttgccatt gcgcagctcc
     2581 tcccgggcgg cctccccggg cgggggcttc ttgacgctct gcttcattag tgggctcttg
     2641 aggaaggcca gcgtcttggc tttcttctcc ttcgggccct cgggccggtg cttgtggact
     2701 cggctgcgac tggccaggga gatgtggatg tacagcaccg tcatgatgac cacaggcagg
     2761 tagaagccag caatggctgt gccaaagcta aacacatgag aaatagagtc ataagcacat
     2821 gggtcacata caggagatat gtatataaca ttaatacaat tttataaaaa aaaggggggg
     2881 cacaactagt ataacttcgt ataaggtata ctatacgaag ttattaagag gtttcatatt
     2941 gctaatagca gctacaatcc agctaccatt ctgcataact tcgtataaag tatcctatac
     3001 gaagttatgt cgactggaga gtggccagcc cttctcatgt tcagagaaca tggttaactg
     3061 gttaagtcat gtcgtcccac aggactcctc cctgcaggac ggcgagttca tctacaaggt
     3121 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     3181 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     3241 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     3301 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     3361 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     3421 caccggcggc atggacgagc tgtacaagta agagctcgaa gttcctatac tatttgaaga
     3481 ataggaactt cttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag
     3541 agaagttcct ataccttttg aagaatagga acttcgaatt cgatatcaag cttatcgata
     3601 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3661 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3721 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3781 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3841 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3901 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3961 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     4021 cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4081 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4141 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag
     4201 cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc
     4261 cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat
     4321 tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa
     4381 ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag
     4441 tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct
     4501 gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt
     4561 gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc
     4621 tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc
     4681 tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa
     4741 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4801 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4861 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4921 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4981 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     5041 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5101 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5161 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5221 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     5281 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     5341 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     5401 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     5461 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5521 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5581 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5641 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5701 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5761 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5821 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5881 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5941 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6001 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6061 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6121 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6181 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6241 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6301 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6361 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6421 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6481 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6541 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6601 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6661 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6721 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6781 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6841 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6901 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6961 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7021 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7081 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7141 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7201 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7261 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7321 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7381 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7441 tcctggcctt ttgctggcct tttgctcaca tgt
//
