LOCUS       sequence-345287-        8922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-HIVNES-GS-dCas13bt3- (GGS)2-huADAR2dd(E488Q) (REPAIR.t3 
            expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8922)
  AUTHORS   Kannan S, Altae-Tran H, Jin X, Madigan VJ, Oshiro R, Makarova KS, 
            Koonin EV, Zhang F
  TITLE     Compact RNA editors with small Cas13 proteins.
  JOURNAL   Nat Biotechnol. 2021 Aug 30. pii: 10.1038/s41587-021-01030-2. doi: 
            10.1038/s41587-021-01030-2.
  PUBMED    34462587
REFERENCE   2  (bases 1 to 8922)
  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: "Nat 
            Biotechnol. 2021 Aug 30. pii: 10.1038/s41587-021-01030-2. doi: 
            10.1038/s41587-021-01030-2."
COMMENT     Sequence Label: pAB1677 REPAIR.t3
FEATURES             Location/Qualifiers
     source          1..8922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      683..692
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             704..730
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(4279..4296)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4285..4509
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4555..4983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4642..4661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4852..4873
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4992..5012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4997..5326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5177..5312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5239..5258
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5393..6187
                     /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(5447..5466)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6057..6076
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6361..6482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6398..6417)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6452..6471
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6531..6547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6531..6547)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6544..6566)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6555..6571
                     /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(6579..6585,6586..6603,6604..6609))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6624..6645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6762..6779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6933..7521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7013..7032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7692..8483,8484..8552))
                     /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     8315..8334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8553..8657)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8732..8751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttgccaccat gggatccctt caactgcctc cacttgaaag
      721 actgacactg ggatccgccc aggtgtccaa gcagaccagc aagaagagag agctgagcat
      781 cgacgagtac cagggcgcca gaaagtggtg cttcacaatc gccttcaaca aggccctggt
      841 caaccgggac aagaacgacg gcctgttcgt ggaaagcctg ctgcggcacg agaagtacag
      901 caagcacgat tggtacgacg aggacacacg ggccctgatc aagtgttcta cccaggccgc
      961 caatgccaag gccgaggctc tggcaaacta cttcagcgcc tacagacaca gccccggctg
     1021 cctgactttt accgccgagg atgagctgcg gaccatcatg gaaagagcct acgagcgggc
     1081 catcttcgag tgcaggcgga gagaaaccga agtgatcatc gagttcccca gcctgttcga
     1141 gggcgacaga attacaacag ccggcgtggt gttcttcgtc agcttctttg tggaacgcag
     1201 agtgctggac agactgtatg gcgccgtgtc cggcctgaag aagaatgagg gccagtacaa
     1261 gctgacccgg aaggccctga gcatgtactg cctgaaggac agcagattca ccaaggcctg
     1321 ggacaagcgg gtgctgctgt tcagagatat tctggctcag ctgggcagaa tccccgccga
     1381 ggcctacgaa tattatcacg gcgagcaggg cgacaagaag cgggccaacg ataacgaggg
     1441 cacaaacccc aagcggcaca aggacaagtt tatcgagttc gccctgcact acctggaagc
     1501 ccagcacagc gagatctgct ttggcagacg gcacatcgtg cgggaagaag ctggcgctgg
     1561 cgacgagcac aagaagcaca gaacaaaggg caaagtggtg gtggacttct ccaagaagga
     1621 cgaggaccag agctactaca tcagcaagaa caacgtgatc gtgcggatcg acaagaatgc
     1681 cggacctcgg agctaccgga tgggcctgaa tgagctgaag tacctggtcc tgctgagcct
     1741 gcaaggcaag ggcgacgatg ccattgccaa gctgtacaga taccggcagc acgtggaaaa
     1801 catcctggac gtggtcaaag tgaccgacaa ggataaccat gtgtttctcc ccagattcgt
     1861 gctggaacag cacggcatcg gcagaaaggc cttcaagcag agaatcgacg gcagagtgaa
     1921 gcacgtgcgc ggcgtgtggg agaagaaaaa ggccgccacc aacgagatga ccctgcacga
     1981 gaaagcccgg gacatcctgc agtacgtgaa cgagaactgc acccgcagct tcaaccccgg
     2041 cgagtacaat agactgctcg tgtgcctcgt gggcaaagac gtcgagaatt tccaggccgg
     2101 gctgaagaga ctgcagctgg ccgagagaat tgatggccgg gtgtacagca tcttcgccca
     2161 gacctccacc atcaatgaga tgcaccaggt cgtgtgcgac cagatcctga accggctgtg
     2221 cagaatcggc gaccagaaac tgtacgacta cgtcggcctg ggcaagaagg atgagatcga
     2281 ctacaagcag aaggtggcct ggttcaaaga gcacatctct atcagacggg gcttcctgcg
     2341 gaagaagttt tggtacgata gcaagaaggg cttcgccaag ctggtcgagg aacacctgga
     2401 atctggcggc ggacagagag atgtgggcct cgataagaag tactaccaca tcgacgccat
     2461 cggaagattt gagggcgcca atcctgctct gtacgagaca ctggccagag acaggctgtg
     2521 cctgatgatg gcccagtact ttctgggcag cgtgcgcaaa gagctgggca acaagatcgt
     2581 gtggtccaac gacagcatcg agctgcctgt ggaaggctcc gtgggcaacg agaaatccat
     2641 cgtgttcagc gtgtccgact acggcaagct gtatgtgctg gacgacgccg agtttctggg
     2701 aagaatctgc gagtacttca tgccccacga gaagggcaag atcagatacc acaccgtgta
     2761 cgagaaaggc ttccgggcct acaacgacct gcagaaaaaa tgcgtggaag ccgtgctggc
     2821 cttcgaggaa aaagtggtca aggccaagaa gatgtccgag aaagagggcg cccactacat
     2881 cgactttcgc gagattctgg cccagaccat gtgcaaagag gccgagaaaa ccgccgtgaa
     2941 caaagtggcg agagccttct ttgcccacca cctgaagttc gtgatcgatg agttcggcct
     3001 gtttagcgac gtgatgaaga agtacggcat cgagaaagaa tggaagttcc ccgtgaaggg
     3061 agggtcaggc ggtagtcagc tgcatttacc gcaggtttta gctgacgctg tctcacgcct
     3121 ggtcctgggt aagtttggtg acctgaccga caacttctcc tcccctcacg ctcgcagaaa
     3181 agtgctggct ggagtcgtca tgacaacagg cacagatgtt aaagatgcca aggtgataag
     3241 tgtttctaca ggaacaaaat gtattaatgg tgaatacatg agtgatcgtg gccttgcatt
     3301 aaatgactgc catgcagaaa taatatctcg gagatccttg ctcagatttc tttatacaca
     3361 acttgagctt tacttaaata acaaagatga tcaaaaaaga tccatctttc agaaatcaga
     3421 gcgagggggg tttaggctga aggagaatgt ccagtttcat ctgtacatca gcacctctcc
     3481 ctgtggagat gccagaatct tctcaccaca tgagccaatc ctggaagaac cagcagatag
     3541 acacccaaat cgtaaagcaa gaggacagct acggaccaaa atagagtctg gtcaggggac
     3601 gattccagtg cgctccaatg cgagcatcca aacgtgggac ggggtgctgc aaggggagcg
     3661 gctgctcacc atgtcctgca gtgacaagat tgcacgctgg aacgtggtgg gcatccaggg
     3721 atcactgctc agcattttcg tggagcccat ttacttctcg agcatcatcc tgggcagcct
     3781 ttaccacggg gaccaccttt ccagggccat gtaccagcgg atctccaaca tagaggacct
     3841 gccacctctc tacaccctca acaagccttt gctcagtggc atcagcaatg cagaagcacg
     3901 gcagccaggg aaggccccca acttcagtgt caactggacg gtaggcgact ccgctattga
     3961 ggtcatcaac gccacgactg ggaaggatga gctgggccgc gcgtcccgcc tgtgtaagca
     4021 cgcgttgtac tgtcgctgga tgcgtgtgca cggcaaggtt ccctcccact tactacgctc
     4081 caagattacc aagcccaacg tgtaccatga gtccaagctg gcggcaaagg agtaccaggc
     4141 cgccaaggcg cgtctgttca cagccttcat caaggcgggg ctgggggcct gggtggagaa
     4201 gcccaccgag caggaccagt tctcactcac gtaagcggcc gctcgagtct agagggcccg
     4261 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     4321 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     4381 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     4441 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     4501 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     4561 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4621 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4681 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4741 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4801 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4861 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4921 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4981 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5041 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5101 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5161 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5221 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     5281 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     5341 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     5401 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     5461 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     5521 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     5581 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     5641 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     5701 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     5761 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5821 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5881 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     5941 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     6001 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     6061 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     6121 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     6181 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     6241 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6301 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     6361 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6421 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6481 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6541 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6601 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6661 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6721 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6781 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6841 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6901 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6961 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7021 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7081 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7141 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     7201 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     7261 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     7321 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     7381 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     7441 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     7501 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     7561 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     7621 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     7681 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     7741 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7801 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7861 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7921 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7981 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     8041 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     8101 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     8161 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     8221 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     8281 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     8341 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     8401 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     8461 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     8521 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     8581 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     8641 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     8701 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     8761 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     8821 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     8881 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
