LOCUS       sequence-209200-        8443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hM3DREADD fused to ABIcs for abscisic acid mediated recruitment of 
            PYLcs labelled proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8443)
  AUTHORS   Roth BL, Gotoh Y, Giguere P, Nichols D
  TITLE     A Chemogenetic Platform for Spatio-temporal Control of beta-arrestin
            Translocation and Signaling at G protein-Coupled Receptors
  JOURNAL   bioRxiv 251769
REFERENCE   2  (bases 1 to 8443)
  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: "bioRxiv 
            251769"
COMMENT     Sequence Label: JGE301_pAAV-hSyn-DIO-Flag-hM3Dq-V2(2D)-ABIcs-m...
FEATURES             Location/Qualifiers
     source          1..8443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   26..155
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        200..647
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(685..718)
                     /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(769..802)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4357..4380)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    4453..4486
                     /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    4537..4570
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4595..5183
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4648..4668)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5066..5077)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5215..5691
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5414..5433)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5731..5871
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5731..5860
                     /label=AAV2 ITR
     rep_origin      5946..6401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6033..6052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6243..6264
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6418..6437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6537..6559
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6597..6615)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6683..6787
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6788..6856,6857..7648)
                     /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(7006..7025)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7819..8407
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8308..8327
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag
       61 gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
      121 agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgtg tgtctagact
      181 gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg
      241 gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa
      301 ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac
      361 tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc
      421 gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg
      481 gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg
      541 agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc
      601 agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc
      661 ggatcctcta gagtcgactc cggaataact tcgtatagga tactttatac gaagttatgc
      721 agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaat aacttcgtat
      781 agcatacatt atacgaagtt atggcgcgcc gaattcctaa ccggtcttgt agagctcatc
      841 catacctccg gtggaatggc ggccttcggc tctctcatat tgttccacga tagtgtagtc
      901 ctcattgtga gaggtgatgt ccagcttgat attgacatta taagcgccag ggagctgcac
      961 aggtttcttg gccttgtatg tggttttgac ctctgcgtcg tagtgacctc cgtctttcaa
     1021 tttcagtctc tgtttgattt ctcctttcag agcgccatcc tcgggataca ttcgttctga
     1081 agatgcctcc cagcccatag tctttttctg cattacgggt ccgtcagagg ggaagttagt
     1141 gccgcgcaat ttcactttat agataaactc accgtcctgc agggaagaat cctgggtcac
     1201 cgtcaccacc cctccatcct cgaaattcat cacccgctcc cacttaaaac cttcggggaa
     1261 agacagtttc aggtagtcag gtatatctgc tgggtgtttg acataggcct tgcttccgta
     1321 cataaattgg ggggagagaa tgtcccaggc aaatgggaga ggacccccct tggtgacttt
     1381 gagtttagca gtttgagtcc cttcatacgg tcgaccttcc ccttcgccct ctatttcgaa
     1441 ctcgtggcca ttgacgctac cctccatatg aactttgaac cgcataaact cctttattat
     1501 ggccatgttg tcctcctctc ccttggacac cataccggtg gacccgcccc cagagccgcc
     1561 acctgtacac ttcagatcca ccacgaccac agagatgtta tccttagacc cccgctgaat
     1621 ggcgagcttt gacaggtatt ctgcggcact catggccgcg gggtccttgc cttccttccg
     1681 tctttcatcg gccagcagag atgcgtcgcc tgccacagcg ttttttttat gccagagcag
     1741 aatccgtttt cttgccattt cacaggcctc ctcgtcagtc atcacgtccc aaactccgtc
     1801 agaagccaga atcagacagt cgtcttcctt aacgcgtttc acggcagtga cttcagggtc
     1861 ggggatgatg gagggtttca gataccgatc gccgatggac ctactcatag caagaacccc
     1921 gaatacgcgg gctccattcc actggatcac cttaccacct gcggcctcga tcctggcggc
     1981 ctcgtcttct cgatcgggct tgtgatccac ggacaagggc agggcagttt tccctcgaca
     2041 cagcactgcc ctggagtctc cacagttggc cacaaagata tgtgaaggga acacgactgc
     2101 cactacagaa gtgctcccca cggtctcggg agccacgctt tcaatctctg agtccactcg
     2161 caaaaaactg ttgaagagcg cttttttcca cttttccagc caggtgtctc cgtcacagag
     2221 catcggcttt tctttagcta tttcctctgc gagagcgaga tgcatgcgct ccctgcagta
     2281 attagccacc tgagatccgc catgcccgtc ataaacgccg aaaaagtgag cagctgactg
     2341 aggatcaaat cggccgtcaa gcatagaccc ggagctggac tgaaggaatc gtggaatggt
     2401 agaaacggca gcctccattt ccggtcttct cccgcagatg gacgtgaatc catagagggg
     2461 cactgtacag cttccccccc cacttccccc tccatcgatc gatgaagtgt ccttggccag
     2521 ggaggagctg gcggtggtgc aggactcatc ttggggaccc aggtcgggtg ggtcgcgtcc
     2581 cagggcttgt tcgggggctc gcttgtggaa aatcacggac tgtctctgct ggtactgctg
     2641 cttccgtctc tttttcttgt cgcattggca caaaaggagc atcttgaatg tcgtcctgaa
     2701 ggttttgttg cacaaggcgt aacaaacagg attcacggta gagttgatgt aacacagcca
     2761 gtagccaaga ttccagaagg ttttaggaat acaggaatca caaaatgtgt tcacgagcac
     2821 catgatgttg taaggggtcc atgttatgat gaatgccaac aggattgcac taagtgtttg
     2881 ggcggctttt ttttccttga ccaggctcat cctttttctc ttagtgatct gggaccgggt
     2941 cttcagcgcg aacctctttg cgagagttgc ttccttaaag gacaagggca gggtagcggt
     3001 agatttgccg acgctgctgt tcacgtcact agtttttgca gtgtctacag cagactcgag
     3061 ttgaattggc agcttagaga aagacttggg gaaactacct ccatcatcaa cggatttctg
     3121 ggcctgcagt ttatctgctt tgcgctcgag gtcgaccatg cccagttcct cctcgggcac
     3181 ctggaggtta tcgctgctgg gaagcttagt tgagttcagg atagtggaat gtcctggcaa
     3241 tttcagcaca atactgtaga tagcgcgggt ttcactgccg atatcctctt cgtcagagct
     3301 tgcggagttt tccagggaag cggcagcgtc gttattattc caggagtcac tacttgagtg
     3361 atcctggtcc atctgttctg agctgggttt ccaagacttg gtggtgaacc aaaaatgaca
     3421 ccttccatat ttccgccggt tggagcgctt catggactgc tgttggagct cgtatgatga
     3481 gcaagatcga gaagagccgg tgggatgcac aaagttctct gtttccgcct ccgtcccgct
     3541 tgcttgaagg ccggccagct ctttagtccg cttctcagtc tctttgtaga tgcgccaata
     3601 aaggatggtc atgatggtga caggcatgta aaagcccgcg attgcagtac cgaaggtgat
     3661 agtaggttcg gacaagaact gtataaaaca ctcgccgggg ggcacggtcc gcttcccaac
     3721 gaaatactgc cagaacagga tagctggggc ccagagtacg aaggaaatca cccaggccag
     3781 gccaatcata actccagccc gtttggttgt cctcttagct cgataggtca aaggcctggt
     3841 aatggagaaa tagcgatcaa aagatatgac gaggagattc atgactgaag cgtttgaggc
     3901 cacacagtct atggccagcc agaggtcgca cgcgaggttt cccagtgccc agcggttcat
     3961 aatgatgtag gtggtaaaga gattcataga gatcactccg atgatcaagt cagcgcacgc
     4021 caggctcagc aggaagtaat tattcacggt tttaagctgc ttattgactt taaaggagac
     4081 aatcaccagg atgttcccta ttatcgttac gagagccaaa atgccagtga ggaaagcaat
     4141 gaaaaccacc tgccaaacgg tgtgccctcc cagtgggtca tcagtggtgc cgtcggggga
     4201 ggaaaaattt ccagctgccc ttgaaacatt ataggagccg aagtgagtca cggtgcctgg
     4261 aggcaggcct gcgtcggatg ggctatgaat ccaagagctg gagatattgg gaaagagtgg
     4321 actggtcgta gaattattgt gaagagtcat atcgatcttg tcatcatcgt ctttatagtc
     4381 ggcgaaaacc aagcagaaaa tataggacag agcgatgata gttttcatgg tggctctaga
     4441 ggatccgcta gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag
     4501 aaattatcac tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat
     4561 acgaagttat gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     4621 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4681 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4741 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4801 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4861 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4921 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4981 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     5041 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5101 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5161 ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg
     5221 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     5281 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     5341 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     5401 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     5461 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     5521 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     5581 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     5641 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     5701 taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc
     5761 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     5821 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     5881 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     5941 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6001 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     6061 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6121 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     6181 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6241 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     6301 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     6361 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     6421 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     6481 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     6541 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     6601 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     6661 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     6721 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     6781 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     6841 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6901 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6961 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     7021 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     7081 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     7141 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     7201 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     7261 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     7321 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     7381 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     7441 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     7501 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     7561 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     7621 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     7681 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7741 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7801 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7861 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7921 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7981 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     8041 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     8101 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     8161 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     8221 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     8281 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     8341 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     8401 atggaaaaac gccagcaacg cggccttttt acggttcctg gcc
//
