LOCUS       sequence-201553-        5774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  b2AR-SPARK protease component.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5774)
  AUTHORS   Kim MW, Wang W, Sanchez MI, Coukos R, von Zastrow M, Ting AY
  TITLE     Time-gated detection of protein-protein interactions with 
            transcriptional readout.
  JOURNAL   Elife. 2017 Nov 30;6. doi: 10.7554/eLife.30233.
  PUBMED    29189201
REFERENCE   2  (bases 1 to 5774)
  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: "Elife."; 
            date: "2017-11-30"; volume: "6. doi"; pages: " 10.7554/eLife.30233"
COMMENT     Sequence Label: pAAV-beta-arrestin2-HA-TEV219
FEATURES             Location/Qualifiers
     source          1..5774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        505..708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     659..679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     753..772
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        753..771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      785..794
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2021..2047
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2048..2074
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2795..2812)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2801..3008
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3037..3177
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3037..3166
                     /label=AAV2 ITR
     rep_origin      3252..3707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3339..3358)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3549..3570
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3724..3743)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3843..3865
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3903..3921)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3989..4093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4094..4162,4163..4954)
                     /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(4312..4331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5125..5713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5614..5633
                     /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 gcgttgtaca tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      781 gcttgccacc atgggtgaaa aacccgggac cagggtcttc aagaagtcga gccctaactg
      841 caagctcacc gtgtacttgg gcaagcgtga ctttgtggat cacttggaca aagtggatcc
      901 tgtcgatggt gtggtgcttg tggatcctga ctacttgaag gaccggaaag tgtttgtgac
      961 cctcacctgt gccttccgct atggccgaga agacctggat gtactgggcc tgtctttccg
     1021 caaagatctg ttcatcgcca cctaccaggc cttccccccc atgcccaacc cacctcggcc
     1081 ccccacccgc ctacaggacc gactgctgaa gaagttgggc cagcacgccc accccttttt
     1141 tttcacaata ccccagaatt tgccttgctc cgtcacactg cagccaggac cggaggacac
     1201 agggaaggcc tgtggagtag actttgagat tcgagccttc tgtgccaaat ctatagaaga
     1261 aaaaagccac aaaaggaact ccgtgcggct tatcatcaga aaggtacagt ttgctcctga
     1321 gacacccggc ccccagccat cagctgaaac cacacgccac ttcctcatgt ctgaccggag
     1381 gtccctgcac ctagaggctt ccctggacaa agagctgtac taccatgggg aacccctcaa
     1441 tgtcaacgtc cacgtcacca acaattctgc caaaaccgtc aagaagatca gagtgtctgt
     1501 gagacagtat gccgacattt gcctcttcag caccgcgcag tacaagtgtc ctgtggctca
     1561 gcttgaacaa gatgaccagg tgtctcccag ttccacattc tgcaaggtgt acaccataac
     1621 cccgctgctc agtgacaacc gagagaagcg tggccttgcc cttgatgggc aactcaagca
     1681 cgaagacacc aacctggctt ccagcaccat tgtgaaggag ggagccaaca aggaggtgct
     1741 gggaatccta gtatcctaca gggtcaaggt gaagctggtg gtgtctcgag gcggggatgt
     1801 ctccgtggag ctacctttcg tcctaatgca ccccaagccc cacgaccaca tcacccttcc
     1861 ccgaccccag tcagcccccc gggaaataga catccctgtg gataccaacc tcattgaatt
     1921 cgataccaac tatgccacag acgacgacat cgtgtttgag gactttgcga ggcttcggct
     1981 gaaggggatg aaggatgacg actgtgatga ccagttctgc tacccatacg atgtgccaga
     2041 ttacgcctat ccctatgacg tccctgatta cgctggtgga agtggatcag gcagcggtgg
     2101 atctggagaa agcttgttta agggaccacg tgattacaac ccgatatcga gcaccatttg
     2161 tcatttgacg aatgaatctg atgggcacac aacatcgttg tatggtattg gatttggtcc
     2221 cttcatcatt acaaacaagc acttgtttag aagaaataat ggaacactgt tggtccaatc
     2281 actacatggt gtattcaagg tcaagaacac cacgactttg caacaacacc tcattgatgg
     2341 gagggacatg ataattattc gcatgcctaa ggatttccca ccatttcctc aaaagctgaa
     2401 atttagagag ccacaaaggg aagagcgcat atgtcttgtg acaaccaact tccaaactaa
     2461 gagcatgtct agcatggtgt cagacactag ttgcacattc ccttcatctg atggcatatt
     2521 ctggaagcat tggattcaaa ccaaggatgg gcagtgtggc agtccattag tatcaactag
     2581 agatgggttc attgttggta tacactcagc atcgaatttc accaacacaa acaattattt
     2641 cacaagcgtg ccgaaaaact tcatggaatt gttgacaaat caggaggcgc agcagtgggt
     2701 tagtggttgg cgattaaatg ctgactcagt attgtggggt ggccataaag ttttcatggt
     2761 gtaagaattc ctcgagctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     2821 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2881 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2941 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3001 gctggggagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     3061 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3121 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3241 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3301 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3361 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3421 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3481 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3541 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3601 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3661 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3721 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3781 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3841 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3901 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3961 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4021 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4081 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4141 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4201 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4261 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4321 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4381 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4441 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4501 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4561 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4621 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4681 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4741 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4801 tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4861 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4921 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4981 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5041 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5101 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5161 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5221 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5281 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5341 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5401 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5461 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5521 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5581 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5641 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5701 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5761 ttttgctcac atgt
//
