LOCUS       sequence-209198-        7444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  beta arrestin 2 fused to PYLcs for abscisic acid mediated 
            recruitment of ABIcs labelled proteins..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7444)
  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 7444)
  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: JGE302_pAAV-hSyn-DIO-Myc-PYLcs-bArrestin2-YFP ...
FEATURES             Location/Qualifiers
     source          1..7444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        180..627
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(665..698)
                     /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(749..782)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3377..3406)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3406..3415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3434..3467
                     /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    3518..3551
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3576..4164
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3629..3649)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4047..4058)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4196..4672
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4395..4414)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4712..4852
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4712..4841
                     /label=AAV2 ITR
     rep_origin      4927..5382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5014..5033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5224..5245
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5399..5418)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5518..5540
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5578..5596)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5664..5768
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5769..5837,5838..6629)
                     /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(5987..6006)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6800..7388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7289..7308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga
      181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac
      241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag
      301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg
      361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg
      421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc
      481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc
      541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga
      601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggtacc ggatcctcta gagtcgactc
      661 cggaataact tcgtatagga tactttatac gaagttatgc agaatggtag ctggattgta
      721 gctgctatta gcaatatgaa acctcttaat aacttcgtat agcatacatt atacgaagtt
      781 atggcgcgcc gaattctcat gtacacttgt agagttcgtc catcccgtgg gtgattccgg
      841 ctgctgtaac aaattccaac aggaccatgt ggtccctttt ctcgttaggg tctttagaca
      901 aggcactctg gtaggacaaa taatggttat cgggcagcag tacgggtcca tcgccaatag
      961 gcgtgttctg ttggtaatga tcggccagct gcacacttcc atcctcgata ttgtgcctga
     1021 ttttgaaatt cactttgatc ccattcttct gcttgtcggc cataatatag acgttgtgag
     1081 aattgtagtt gtactccagt ttgtggccga gaatattccc atcctctttg aagtcgattc
     1141 ccttgagttc gatgcggttg acaagcgtat caccttcaaa tttgacctcg gctcttgtct
     1201 tgtagtttcc gtcgtcctta aagaagattg tgcgttcctg gacatatcct tcgggcatag
     1261 cgcttttgaa gaaatcatgc cgcttcatat ggtcaggata ccgggcaaaa cactgcaggc
     1321 cgtatccgaa ggttgtgacc agcgtgggcc atggcacagg gagcttaccc gtagtgcaga
     1381 taaatttcag cgtcaatttt ccgtaggtgg cgtccccctc tccctcaccg cttacggaaa
     1441 acttgtgacc gttcacgtcg ccatcgagtt ctaccaagat aggcaccacg cctgtgaaca
     1501 gctcttcgcc cttactcatt gtacaagacc cgccgccgga cccgccccca tcgatgcaca
     1561 gctggtcgtc ataatcgtcg tccttcatgc ccttcagtct aaggcgtgca aagtcttcaa
     1621 atacgatgtc gtcgtctgtg gcgtagttcg tgtcaaactc gatgaggtta gtgtccactg
     1681 ggacatctgt ctccggtgct gcggactgag gccggggcag aggaatatga tcgtgtggct
     1741 taggatgcat gagaacaaaa gggagttcaa ctgagacgtc ccctccgcga gaaaccacca
     1801 gtttaacctt caccctgtag gacacgagga tgccaaggac ttctttgttt gcgccttcct
     1861 ttacgatggt gctggaagcc agattcgtgt cctcgtgctt gagcttacca tccagagcga
     1921 gcccccgctt ttcccggtta tcgctcagca ggggtgtaat tgtatacacc ttgcaaaatg
     1981 tggaactggg gctaacctgg tcgtcctgtt ccagctgggc gacagggcac ttatactgag
     2041 cagtagagaa cagacatata tcagcgtatt gccttacaga tacctttatc ttcttgaccg
     2101 tcttggtgga gttgttcgtc acatgaacgt tcacattcaa gggctcacca tggtagtaca
     2161 gttccttgtc caaagaggcc tccagatgca aacttctatc tgacataagg aaatgtcttg
     2221 ttgtttccgc tgatggctga gggccaggct tctcaggtgc aaattgcact ttccgtataa
     2281 ccaacctcac gctgttccgc ttgtgtgatt tctcttccag gctcttggca caaaaggcgc
     2341 gtatttcaaa atcgacccca cacgccttcc ccgtgtcctc gggcccaggc tggagggtca
     2401 cgctgcaggg gaggttttgt gggattgtga aaaagaaggg atgggcgtgt tggccgagct
     2461 tcctgagaag cctgtcttgg aggcgggtag gtggtcgagg agggttaggg actggaggga
     2521 aggcctgata tgtggcgata aacaggtcct ttctaaaact caagcccagc acatccaggt
     2581 cctcccgccc atatctaaag gcacaggtca gagtgacaaa gactttccgg tctttcagat
     2641 aatcgggatc gacgagcacc actccatcca ccggatctac cttatccaga tgatctacga
     2701 aatcgcgctt gccgaggtag acggtgagct tgcaattggg actagatttc ttgaacaccc
     2761 tagtgcccgg cttttctccc atatcgatag atgaaccgcc ggcactgccc gctgagcctg
     2821 cgctatcaaa ctcaccggta ttcatagctt ctgtgattga ggcgagtttt tgcaggttca
     2881 gtcgaatcac tgtgtctgca aacaggcggg tgtcctcctc agagtttccc tctggcacgt
     2941 ctaccacata ggattcgagc accacggtcc aaatccgctc ttcctcctcc tctttttcga
     3001 atctatgcac tgtggtcact gatttgtagt tgcgcaagcg gtgttccccg ccggtgatgg
     3061 aaaaccctgt cactctccga tcgtcgtcca gcaggtcgag ccgctccctg gacgtattgg
     3121 ccggcaggcc actgattaca ttaacgtccc tagtgcatcc tacccgcatc tcaaagtctt
     3181 cgctcacgtt acaactcttg atgaaatgct tgtatatctg aggtcggtcg aagcggcgca
     3241 cgacactcca cacagtctca gggggggcat ggatgcgctg agcgaggaga cttgaacacc
     3301 tgccattgcc cagctggtac gtgtgaaatt cagcaattga ctgagagagc tgggtgaact
     3361 cgtcttgtgt accggtcaaa tcctcttcac tgatcagctt ctgttccatg gtggctctag
     3421 aggatccgct agcataactt cgtataaagt atcctatacg aagttatttg ccttaaccca
     3481 gaaattatca ctgttattct ttagaatggt gcaaagaata acttcgtata atgtatgcta
     3541 tacgaagtta tgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     3601 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3661 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3721 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3781 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3841 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3901 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3961 tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc
     4021 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     4081 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     4141 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     4201 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     4261 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     4321 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     4381 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     4441 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     4501 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     4561 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     4621 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     4681 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     4741 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4801 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     4861 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4921 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4981 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     5041 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5101 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     5161 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5221 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     5281 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     5341 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     5401 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     5461 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     5521 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     5581 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5641 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5701 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5761 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5821 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5881 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5941 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6001 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6061 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6121 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6181 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6241 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6301 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6361 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6421 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6481 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6541 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6601 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6661 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6721 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6781 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6841 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6901 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6961 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7021 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7081 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7141 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7201 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7261 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7321 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7381 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7441 ctca
//
