LOCUS       sequence-345282-        7749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-HIVNES-GS-Cas13bt3 (human codon optimized Cas13bt3 expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7749)
  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 7749)
  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: pAB1679 CMV-HIVNES-GS-Cas13bt3
FEATURES             Location/Qualifiers
     source          1..7749
                     /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(3106..3123)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3112..3336
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3382..3810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3469..3488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3679..3700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3819..3839)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3824..4153
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4004..4139
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4066..4085
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4220..5014
                     /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(4274..4293)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4884..4903
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5188..5309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5225..5244)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5279..5298
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5358..5374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5358..5374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5371..5393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5382..5398
                     /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(5406..5412,5413..5430,5431..5436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5451..5472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5589..5606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5760..6348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5840..5859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6519..7310,7311..7379))
                     /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     7142..7161
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7380..7484)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7559..7578)
                     /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 tgagaaacta cttcagccac 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 caaagtgcgg agagccttct ttcaccacca cctgaagttc gtgatcgatg agttcggcct
     3001 gtttagcgac gtgatgaaga agtacggcat cgagaaagaa tggaagttcc ccgtgaagta
     3061 agcggccgct cgagtctaga gggcccgttt aaacccactg atcagcctcg actgtgcctt
     3121 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3181 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3241 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3301 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3361 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3421 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3481 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3541 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3601 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3661 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3721 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3781 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3841 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3901 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3961 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4021 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4081 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4141 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4201 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4261 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4321 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4381 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4441 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4501 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4561 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4621 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4681 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4741 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4801 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4861 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4921 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4981 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5041 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5101 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5161 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5221 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5281 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5341 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5401 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5461 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5521 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5581 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5641 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5701 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5761 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5821 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5881 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5941 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6001 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6061 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6121 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6181 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6241 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6301 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6361 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6421 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6481 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6541 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6601 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6661 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6721 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6781 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6841 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6901 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6961 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7021 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7081 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7141 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7201 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7261 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7321 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7381 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7441 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7501 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     7561 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     7621 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     7681 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     7741 atacgcgtt
//
