LOCUS       sequence-345283-        7836 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-HIVNES-GS-Cas13bt1 (human codon optimized Cas13bt1 expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7836)
  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 7836)
  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: pAB1678 CMV-HIVNES-GS-Cas13bt1
FEATURES             Location/Qualifiers
     source          1..7836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2024..2041)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2030..2254
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2300..2728
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2387..2406)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2597..2618
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2737..2757)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2742..3071
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2922..3057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2984..3003
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3138..3932
                     /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(3192..3211)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3802..3821
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4106..4227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4143..4162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4197..4216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4276..4292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4276..4292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4289..4311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4300..4316
                     /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(4324..4330,4331..4348,4349..4354))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4369..4390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4507..4524)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4678..5266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4758..4777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5437..6228,6229..6297))
                     /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     6060..6079
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6298..6402)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6477..6496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6668..7047
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7048..7251
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7202..7222
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7296..7315
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7296..7314
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      7350..7359
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7371..7397
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
ORIGIN
        1 tgtttgagga agaggacaag atcacctctg ccggcgtggt gttcttcgtc agctttttta
       61 tcgagcggcg gttcctgaac cggctgatgg gctatgtgca gggcttcaga aagacagagg
      121 gcgagtacaa catcaccaga caggtgttca gcaagtactg cctgaaggac agctacagcg
      181 tgcaggccca ggatcacgat gccgtgatgt tcagagacat cctgggctac ctgagcagag
      241 tgcccaccga gatctaccag cacatcaagc tgacccggaa gcggagccag gatcagctga
      301 gcgagagaaa gaccgacaag ttcatcctgt tcgccctgaa gtacctcgag gactacggac
      361 tgaaggacct ggccgattac accgcctgct ttgccagaag caagatcaag agagagaacg
      421 aggacaccaa agagacagac ggcaacaagc acaagttcca ccgcgagaag cccgtggtgg
      481 aaatccactt cgacaaagag aagcaggatc agttctacat caagcggaac aacgtcatcc
      541 tgaaggccca gaagaaaggc ggccagagca acgtgttccg gatgggagtg tacgagctga
      601 agtatctggt gctgctgagc ctgctgggca aagccgaaga ggccatccag agaatcgacc
      661 ggtacatcag cagcctgaag aaacagctgc cctacctgga taagatcagc aacgaagaga
      721 tccagaagtc catcaacttt ctgcccagat tcgtgcggag ccggctggga ctgctgcaag
      781 tggatgatga gaagagactg aaaacccggc tggaatacgt gaaggccaag tggaccgata
      841 agaaagaggg cagcagaaag ctggaactgc accggaaggg cagagatatc ctgcggtata
      901 tcaacgagag atgcgacaga cccctgagcc ggaaagagta taacaacatt ctgaagttca
      961 tcgtgaacaa ggacttcgcc ggcttctaca acgagctgga agaactgaag cggaccagac
     1021 ggctggacaa gaacatcatc cagaagctga gcggccacac cacactgaac gctctgcacg
     1081 agcgagtgtg tgacctggtc ctgcaagagc tgggctctct gcagtccgag aacctgaaag
     1141 agtacatcgg actgatcccc aaagaagaga aagaagtcac cttccgcgag aaagtggaca
     1201 gaatcctgga acagcctgtg gtgtacaagg gcttcctgag atacgagttc ttcaaagaag
     1261 ataagaaatc cttcgccagg ctggtggaag aggctatcaa gaccaagtgg agcgacttcg
     1321 acatccctct gggcgaagag tactacaaca tccctagcct ggacagattc gaccggacca
     1381 acaagaagct gtacgagaca ctggccatgg accggctgtg tctgatgatg gccagacagt
     1441 actacctgcg gctgaacgag aagctggccg agaaggcaca gcacatctac tggaaaaaag
     1501 aggacggccg ggaagtcatc atcttcaagt tccagaatcc gaaagagcag aagaagtcct
     1561 tcagcatcag gttcagcatc ctggactaca ccaagatgta cgtgatggac gaccccgagt
     1621 tcctgagcag gctgtgggag tacttcatcc ctaaagaggc caaagagatc gactaccaca
     1681 agcactacgc cagagccttc gacaagtaca ccaacctgca gaaagaaggc atcgacgcca
     1741 tcctgaaact ggaaggcaga atcatcgagc gccggaagat caagcccgcc aagaactaca
     1801 tcgagtttca agagattatg aatcggagcg gctacaacaa cgaccagcag gttgccctga
     1861 agagagtgcg gaacgccctg ctgcactaca acctgaactt cgagagagag cacctgaagc
     1921 ggttctacgg cgtcgtgaag agagagggca tcgagaaaaa atggtccctg atcgtgtaag
     1981 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     2041 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2101 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2161 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2221 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2281 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2341 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2401 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2461 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2521 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2581 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2641 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2701 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2761 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2821 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2881 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2941 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3001 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3061 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3121 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3181 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3241 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3301 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3361 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3421 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3481 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3541 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3601 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3661 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3721 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3781 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3841 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3901 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3961 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4021 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4081 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4141 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4201 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4261 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4321 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4381 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4441 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4501 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4561 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4861 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4921 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4981 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5041 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5101 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5161 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5521 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5581 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5641 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5701 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5761 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5821 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5881 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5941 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6001 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6061 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6121 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6181 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6241 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6301 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6361 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6421 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     6481 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     6541 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     6601 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     6661 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     6721 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     6781 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     6841 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     6901 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     6961 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     7021 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     7081 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     7141 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     7201 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     7261 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     7321 caagctggct agcgtttaaa cttaagcttg ccaccatggg atcccttcaa ctgcctccac
     7381 ttgaaagact gacactggga tccgaattcg agaacatcaa gaaaaccagc aacaaagagg
     7441 tgtacagcat cgagcagtac gagggcgaga agaagtggtg cttcgccatc gtgctgaaca
     7501 gagcccagac caacctggaa gagaacccca agctgttcga gcagaccctg accagatttg
     7561 agaagatcat gaagcaggac tggttcaacg aggaaacgaa gaagctgatc tacgagaaag
     7621 aggaagaaaa caaagtcaaa gaggaaatcc agatcgccgc cagcgagcgg ctgaagaacc
     7681 tgagaaacta cttcagccac tacctgcacg cccctgactg cctgatcttc aaccggaacg
     7741 acaccatccg gatcatcatg gaaaaggcct acgagaagtc ccgcttcgag gccaagaaga
     7801 agcagcaaga ggacatctcc atcgagttcc ccgagc
//
