LOCUS       sequence-202297-        7455 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 7455)
  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 7455)
  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..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(671..704)
                     /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(755..788)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3383..3412)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3412..3421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3440..3473
                     /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    3524..3557
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3582..4170
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3635..3655)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4053..4064)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4202..4678
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4401..4420)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4718..4858
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4718..4847
                     /label=AAV2 ITR
     rep_origin      4933..5388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5020..5039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5230..5251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5405..5424)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5524..5546
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5584..5602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5670..5774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5775..5843,5844..6635)
                     /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(5993..6012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6806..7394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7295..7314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggtaccggat cctctagagt
      661 cgactccgga ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg
      721 attgtagctg ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac
      781 gaagttatgg cgcgccgaat tctcatgtac acttgtagag ttcgtccatc ccgtgggtga
      841 ttccggctgc tgtaacaaat tccaacagga ccatgtggtc ccttttctcg ttagggtctt
      901 tagacaaggc actctggtag gacaaataat ggttatcggg cagcagtacg ggtccatcgc
      961 caataggcgt gttctgttgg taatgatcgg ccagctgcac acttccatcc tcgatattgt
     1021 gcctgatttt gaaattcact ttgatcccat tcttctgctt gtcggccata atatagacgt
     1081 tgtgagaatt gtagttgtac tccagtttgt ggccgagaat attcccatcc tctttgaagt
     1141 cgattccctt gagttcgatg cggttgacaa gcgtatcacc ttcaaatttg acctcggctc
     1201 ttgtcttgta gtttccgtcg tccttaaaga agattgtgcg ttcctggaca tatccttcgg
     1261 gcatagcgct tttgaagaaa tcatgccgct tcatatggtc aggataccgg gcaaaacact
     1321 gcaggccgta tccgaaggtt gtgaccagcg tgggccatgg cacagggagc ttacccgtag
     1381 tgcagataaa tttcagcgtc aattttccgt aggtggcgtc cccctctccc tcaccgctta
     1441 cggaaaactt gtgaccgttc acgtcgccat cgagttctac caagataggc accacgcctg
     1501 tgaacagctc ttcgccctta ctcattgtac aagacccgcc gccggacccg cccccatcga
     1561 tgcacagctg gtcgtcataa tcgtcgtcct tcatgccctt cagtctaagg cgtgcaaagt
     1621 cttcaaatac gatgtcgtcg tctgtggcgt agttcgtgtc aaactcgatg aggttagtgt
     1681 ccactgggac atctgtctcc ggtgctgcgg actgaggccg gggcagagga atatgatcgt
     1741 gtggcttagg atgcatgaga acaaaaggga gttcaactga gacgtcccct ccgcgagaaa
     1801 ccaccagttt aaccttcacc ctgtaggaca cgaggatgcc aaggacttct ttgtttgcgc
     1861 cttcctttac gatggtgctg gaagccagat tcgtgtcctc gtgcttgagc ttaccatcca
     1921 gagcgagccc ccgcttttcc cggttatcgc tcagcagggg tgtaattgta tacaccttgc
     1981 aaaatgtgga actggggcta acctggtcgt cctgttccag ctgggcgaca gggcacttat
     2041 actgagcagt agagaacaga catatatcag cgtattgcct tacagatacc tttatcttct
     2101 tgaccgtctt ggtggagttg ttcgtcacat gaacgttcac attcaagggc tcaccatggt
     2161 agtacagttc cttgtccaaa gaggcctcca gatgcaaact tctatctgac ataaggaaat
     2221 gtcttgttgt ttccgctgat ggctgagggc caggcttctc aggtgcaaat tgcactttcc
     2281 gtataaccaa cctcacgctg ttccgcttgt gtgatttctc ttccaggctc ttggcacaaa
     2341 aggcgcgtat ttcaaaatcg accccacacg ccttccccgt gtcctcgggc ccaggctgga
     2401 gggtcacgct gcaggggagg ttttgtggga ttgtgaaaaa gaagggatgg gcgtgttggc
     2461 cgagcttcct gagaagcctg tcttggaggc gggtaggtgg tcgaggaggg ttagggactg
     2521 gagggaaggc ctgatatgtg gcgataaaca ggtcctttct aaaactcaag cccagcacat
     2581 ccaggtcctc ccgcccatat ctaaaggcac aggtcagagt gacaaagact ttccggtctt
     2641 tcagataatc gggatcgacg agcaccactc catccaccgg atctacctta tccagatgat
     2701 ctacgaaatc gcgcttgccg aggtagacgg tgagcttgca attgggacta gatttcttga
     2761 acaccctagt gcccggcttt tctcccatat cgatagatga accgccggca ctgcccgctg
     2821 agcctgcgct atcaaactca ccggtattca tagcttctgt gattgaggcg agtttttgca
     2881 ggttcagtcg aatcactgtg tctgcaaaca ggcgggtgtc ctcctcagag tttccctctg
     2941 gcacgtctac cacataggat tcgagcacca cggtccaaat ccgctcttcc tcctcctctt
     3001 tttcgaatct atgcactgtg gtcactgatt tgtagttgcg caagcggtgt tccccgccgg
     3061 tgatggaaaa ccctgtcact ctccgatcgt cgtccagcag gtcgagccgc tccctggacg
     3121 tattggccgg caggccactg attacattaa cgtccctagt gcatcctacc cgcatctcaa
     3181 agtcttcgct cacgttacaa ctcttgatga aatgcttgta tatctgaggt cggtcgaagc
     3241 ggcgcacgac actccacaca gtctcagggg gggcatggat gcgctgagcg aggagacttg
     3301 aacacctgcc attgcccagc tggtacgtgt gaaattcagc aattgactga gagagctggg
     3361 tgaactcgtc ttgtgtaccg gtcaaatcct cttcactgat cagcttctgt tccatggtgg
     3421 ctctagagga tccgctagca taacttcgta taaagtatcc tatacgaagt tatttgcctt
     3481 aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt
     3541 atgctatacg aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca
     3601 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3661 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3721 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3781 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3841 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3901 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3961 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     4021 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     4081 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     4141 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     4201 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     4261 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     4321 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     4381 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     4441 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     4501 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     4561 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     4621 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     4681 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     4741 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4801 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     4861 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     4921 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4981 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     5041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5221 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     5281 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     5401 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     5461 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     5521 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     5581 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5641 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5701 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5761 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5821 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5881 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5941 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6001 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6061 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6121 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6181 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6241 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6301 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6361 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6421 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6481 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6541 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6601 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6661 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6721 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6781 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6841 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6901 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6961 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7021 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7081 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7141 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7201 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7261 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7321 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7381 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7441 cttttgctca catgt
//
