LOCUS       sequence-377069-        9036 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Express full length ADAR1p150 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9036)
  AUTHORS   Jiang K, Koob J, Chen XD, Krajeski RN, Zhang Y, Volf V, Zhou W, 
            Sgrizzi SR, Villiger L, Gootenberg JS, Chen F, Abudayyeh OO
  TITLE     Programmable eukaryotic protein synthesis with RNA sensors by 
            harnessing ADAR.
  JOURNAL   Nat Biotechnol. 2022 Oct 27. pii: 10.1038/s41587-022-01534-5. doi: 
            10.1038/s41587-022-01534-5.
  PUBMED    36302988
REFERENCE   2  (bases 1 to 9036)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2022 Oct 27. pii: 10.1038/s41587-022-01534-5. doi: 
            10.1038/s41587-022-01534-5."
COMMENT     Sequence Label: pDY1173_ADAR1p150
FEATURES             Location/Qualifiers
     source          1..9036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        155..534
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        535..738
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     689..709
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     783..802
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        783..801
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4547..4564)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4553..4777
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4823..5251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4910..4929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5120..5141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5260..5280)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5265..5594
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5445..5580
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5507..5526
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5661..6455
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5715..5734)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6325..6344
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6629..6750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6666..6685)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6720..6739
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6799..6815)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6799..6815)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6812..6834)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6823..6839
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6847..6853,6854..6871,6872..6877))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6892..6913
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7030..7047)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7201..7789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7281..7300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7960..8751,8752..8820))
                     /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     8583..8602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8821..8925)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9000..9019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
       61 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
      121 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
      181 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      241 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      301 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      361 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      421 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      481 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      541 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      601 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      661 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      721 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
      781 attaatacga ctcactatag ggagacccaa gctggctagc ccaccatgaa gcttatgaat
      841 ccgcggcagg ggtattccct cagcggatac tacacccatc catttcaagg ctatgagcac
      901 agacagctca gataccagca gcctgggcca ggatcttccc ccagtagttt cctgcttaag
      961 caaatagaat ttctcaaggg gcagctccca gaagcaccgg tgattggaaa gcagacaccg
     1021 tcactgccac cttccctccc aggactccgg ccaaggtttc cagtactact tgcctccagt
     1081 accagaggca ggcaagtgga catcaggggt gtccccaggg gcgtgcatct cggaagtcag
     1141 gggctccaga gagggttcca gcatccttca ccacgtggca ggagtctgcc acagagaggt
     1201 gttgattgcc tttcctcaca tttccaggaa ctgagtatct accaagatca ggaacaaagg
     1261 atcttaaagt tcctggaaga gcttggggaa gggaaggcca ccacagcaca tgatctgtct
     1321 gggaaacttg ggactccgaa gaaagaaatc aatcgagttt tatactccct ggcaaagaag
     1381 ggcaagctac agaaagaggc aggaacaccc cctttgtgga aaatcgcggt ctccactcag
     1441 gcttggaacc agcacagcgg agtggtaaga ccagacggtc atagccaagg agccccaaac
     1501 tcagacccga gtttggaacc ggaagacaga aactccacat ctgtctcaga agatcttctt
     1561 gagcctttta ttgcagtctc agctcaggct tggaaccagc acagcggagt ggtaagacca
     1621 gacagtcata gccaaggatc cccaaactca gacccaggtt tggaacctga agacagcaac
     1681 tccacatctg ccttggaaga tcctcttgag tttttagaca tggccgagat caaggagaaa
     1741 atctgcgact atctcttcaa tgtgtctgac tcctctgccc tgaatttggc taaaaatatt
     1801 ggccttacca aggcccgaga tataaatgct gtgctaattg acatggaaag gcagggggat
     1861 gtctatagac aagggacaac ccctcccata tggcatttga cagacaagaa gcgagagagg
     1921 atgcaaatca agagaaatac gaacagtgtt cctgaaaccg ctccagctgc aatccctgag
     1981 accaaaagaa acgcagagtt cctcacctgt aatataccca catcaaatgc ctcaaataac
     2041 atggtaacca cagaaaaagt ggagaatggg caggaacctg tcataaagtt agaaaacagg
     2101 caagaggcca gaccagaacc agcaagactg aaaccacctg ttcattacaa tggcccctca
     2161 aaagcagggt atgttgactt tgaaaatggc cagtgggcca cagatgacat cccagatgac
     2221 ttgaatagta tccgcgcagc accaggtgag tttcgagcca tcatggagat gccctccttc
     2281 tacagtcatg gcttgccacg gtgttcaccc tacaagaaac tgacagagtg ccagctgaag
     2341 aaccccatca gcgggctgtt agaatatgcc cagttcgcta gtcaaacctg tgagttcaac
     2401 atgatagagc agagtggacc accccatgaa cctcgattta aattccaggt tgtcatcaat
     2461 ggccgagagt ttcccccagc tgaagctgga agcaagaaag tggccaagca ggatgcagct
     2521 atgaaagcca tgacaattct gctagaggaa gccaaagcca aggacagtgg aaaatcagaa
     2581 gaatcatccc actattccac agagaaagaa tcagagaaga ctgcagagtc ccagaccccc
     2641 accccttcag ccacatcctt cttttctggg aagagccccg tcaccacact gcttgagtgt
     2701 atgcacaaat tggggaactc ctgcgaattc cgtctcctgt ccaaagaagg ccctgcccat
     2761 gaacccaagt tccaatactg tgttgcagtg ggagcccaaa ctttccccag tgtgagtgct
     2821 cccagcaaga aagtggcaaa gcagatggcc gcagaggaag ccatgaaggc cctgcatggg
     2881 gaggcgacca actccatggc ttctgataac cagcctgaag gtatgatctc agagtcactt
     2941 gataacttgg aatccatgat gcccaacaag gtcaggaaga ttggcgagct cgtgagatac
     3001 ctgaacacca accctgtggg tggccttttg gagtacgccc gctcccatgg ctttgctgct
     3061 gaattcaagt tggtcgacca gtccggacct cctcacgagc ccaagttcgt ttaccaagca
     3121 aaagttgggg gtcgctggtt cccagccgtc tgcgcacaca gcaagaagca aggcaagcag
     3181 gaagcagcag atgcggctct ccgtgtcttg attggggaga acgagaaggc agaacgcatg
     3241 ggtttcacag aggtaacccc agtgacaggg gccagtctca gaagaactat gctcctcctc
     3301 tcaaggtccc cagaagcaca gccaaagaca ctccctctca ctggcagcac cttccatgac
     3361 cagatagcca tgctgagcca ccggtgcttc aacactctga ctaacagctt ccagccctcc
     3421 ttgctcggcc gcaagattct ggccgccatc attatgaaaa aagactctga ggacatgggt
     3481 gtcgtcgtca gcttgggaac agggaatcgc tgtgtaaaag gagattctct cagcctaaaa
     3541 ggagaaactg tcaatgactg ccatgcagaa ataatctccc ggagaggctt catcaggttt
     3601 ctctacagtg agttaatgaa atacaactcc cagactgcga aggatagtat atttgaacct
     3661 gctaagggag gagaaaagct ccaaataaaa aagactgtgt cattccatct gtatatcagc
     3721 actgctccgt gtggagatgg cgccctcttt gacaagtcct gcagcgaccg tgctatggaa
     3781 agcacagaat cccgccacta ccctgtcttc gagaatccca aacaaggaaa gctccgcacc
     3841 aaggtggaga acggagaagg cacaatccct gtggaatcca gtgacattgt gcctacgtgg
     3901 gatggcattc ggctcgggga gagactccgt accatgtcct gtagtgacaa aatcctacgc
     3961 tggaacgtgc tgggcctgca aggggcactg ttgacccact tcctgcagcc catttatctc
     4021 aaatctgtca cattgggtta ccttttcagc caagggcatc tgacccgtgc tatttgctgt
     4081 cgtgtgacaa gagatgggag tgcatttgag gatggactac gacatccctt tattgtcaac
     4141 caccccaagg ttggcagagt cagcatatat gattccaaaa ggcaatccgg gaagactaag
     4201 gagacaagcg tcaactggtg tctggctgat ggctatgacc tggagatcct ggacggtacc
     4261 agaggcactg tggatgggcc acggaatgaa ttgtcccggg tctccaaaaa gaacattttt
     4321 cttctattta agaagctctg ctccttccgt taccgcaggg atctactgag actctcctat
     4381 ggtgaggcca agaaagctgc ccgtgactac gagacggcca agaactactt caaaaaaggc
     4441 ctgaaggata tgggctatgg gaactggatt agcaaacccc aggaggaaaa gaacttttat
     4501 ctctgcccag tatagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     4561 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4621 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4681 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4741 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4801 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4861 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4921 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4981 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5041 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5101 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5161 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5221 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     5281 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5341 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5401 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5461 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5521 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5581 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     5641 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5701 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5761 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5821 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5881 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5941 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     6001 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     6061 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6121 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6181 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6241 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6301 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6361 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6421 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6481 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6541 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6601 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     6661 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     6721 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     6781 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     6841 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6901 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6961 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7021 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7081 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7141 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7201 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7261 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7321 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7381 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7441 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7501 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7561 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7621 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     7681 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7741 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7801 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7861 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7921 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8041 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8161 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8401 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8461 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8641 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8821 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8881 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8941 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     9001 acaatctgct ctgatgccgc atagttaagc cagtat
//
