LOCUS       sequence-345279-        9009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV-HIVNES-GS-dCas13bt1- (GGS)2-huADAR2dd(E488Q) (REPAIR.t1 
            expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9009)
  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 9009)
  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: pAB1676 REPAIR.t1
FEATURES             Location/Qualifiers
     source          1..9009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(123..143)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        128..457
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      308..443
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     370..389
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             524..1318
                     /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(578..597)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1188..1207
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1492..1613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1529..1548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1583..1602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1662..1678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1662..1678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1675..1697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1686..1702
                     /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(1710..1716,1717..1734,1735..1740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1755..1776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1893..1910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2064..2652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2144..2163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2823..3614,3615..3683))
                     /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     3446..3465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3684..3788)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3863..3882)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4054..4433
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4434..4637
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4588..4608
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4682..4701
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4682..4700
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4736..4745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4757..4783
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(8419..8436)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8425..8649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8695..9009,1..114)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8782..8801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(8992..9009,1..4)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
       61 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
      121 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      181 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
      241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
      301 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
      361 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
      421 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
      481 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
      541 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
      601 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
      661 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
      721 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
      781 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
      841 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
      901 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
      961 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     1021 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     1081 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     1141 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     1201 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     1261 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     1321 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     1381 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     1441 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     1501 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1561 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1621 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     1681 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1741 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1801 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1861 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1921 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1981 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2041 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2701 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2761 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2821 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2881 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2941 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3001 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3061 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3121 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3181 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3241 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3301 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3361 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3421 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3481 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3541 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3601 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3661 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3721 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3781 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     3841 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     3901 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     3961 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     4021 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     4081 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     4141 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     4201 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     4261 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     4321 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     4381 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     4441 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     4501 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     4561 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     4621 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     4681 ttaatacgac tcactatagg gagacccaag ctggctagcg tttaaactta agcttgccac
     4741 catgggatcc cttcaactgc ctccacttga aagactgaca ctgggatccg aattcgagaa
     4801 catcaagaaa accagcaaca aagaggtgta cagcatcgag cagtacgagg gcgagaagaa
     4861 gtggtgcttc gccatcgtgc tgaacagagc ccagaccaac ctggaagaga accccaagct
     4921 gttcgagcag accctgacca gatttgagaa gatcatgaag caggactggt tcaacgagga
     4981 aacgaagaag ctgatctacg agaaagagga agaaaacaaa gtcaaagagg aaatccagat
     5041 cgccgccagc gagcggctga agaacctggc aaactacttc agcgcctacc tgcacgcccc
     5101 tgactgcctg atcttcaacc ggaacgacac catccggatc atcatggaaa aggcctacga
     5161 gaagtcccgc ttcgaggcca agaagaagca gcaagaggac atctccatcg agttccccga
     5221 gctgtttgag gaagaggaca agatcacctc tgccggcgtg gtgttcttcg tcagcttttt
     5281 tatcgagcgg cggttcctga accggctgat gggctatgtg cagggcttca gaaagacaga
     5341 gggcgagtac aacatcacca gacaggtgtt cagcaagtac tgcctgaagg acagctacag
     5401 cgtgcaggcc caggatcacg atgccgtgat gttcagagac atcctgggct acctgagcag
     5461 agtgcccacc gagatctacc agcacatcaa gctgacccgg aagcggagcc aggatcagct
     5521 gagcgagaga aagaccgaca agttcatcct gttcgccctg aagtacctcg aggactacgg
     5581 actgaaggac ctggccgatt acaccgcctg ctttgccaga agcaagatca agagagagaa
     5641 cgaggacacc aaagagacag acggcaacaa gcacaagttc caccgcgaga agcccgtggt
     5701 ggaaatccac ttcgacaaag agaagcagga tcagttctac atcaagcgga acaacgtcat
     5761 cctgaaggcc cagaagaaag gcggccagag caacgtgttc cggatgggag tgtacgagct
     5821 gaagtatctg gtgctgctga gcctgctggg caaagccgaa gaggccatcc agagaatcga
     5881 ccggtacatc agcagcctga agaaacagct gccctacctg gataagatca gcaacgaaga
     5941 gatccagaag tccatcaact ttctgcccag attcgtgcgg agccggctgg gactgctgca
     6001 agtggatgat gagaagagac tgaaaacccg gctggaatac gtgaaggcca agtggaccga
     6061 taagaaagag ggcagcagaa agctggaact gcaccggaag ggcagagata tcctgcggta
     6121 tatcaacgag agatgcgaca gacccctgag ccggaaagag tataacaaca ttctgaagtt
     6181 catcgtgaac aaggacttcg ccggcttcta caacgagctg gaagaactga agcggaccag
     6241 acggctggac aagaacatca tccagaagct gagcggccac accacactga acgctctgca
     6301 cgagcgagtg tgtgacctgg tcctgcaaga gctgggctct ctgcagtccg agaacctgaa
     6361 agagtacatc ggactgatcc ccaaagaaga gaaagaagtc accttccgcg agaaagtgga
     6421 cagaatcctg gaacagcctg tggtgtacaa gggcttcctg agatacgagt tcttcaaaga
     6481 agataagaaa tccttcgcca ggctggtgga agaggctatc aagaccaagt ggagcgactt
     6541 cgacatccct ctgggcgaag agtactacaa catccctagc ctggacagat tcgaccggac
     6601 caacaagaag ctgtacgaga cactggccat ggaccggctg tgtctgatga tggccagaca
     6661 gtactacctg cggctgaacg agaagctggc cgagaaggca cagcacatct actggaaaaa
     6721 agaggacggc cgggaagtca tcatcttcaa gttccagaat ccgaaagagc agaagaagtc
     6781 cttcagcatc aggttcagca tcctggacta caccaagatg tacgtgatgg acgaccccga
     6841 gttcctgagc aggctgtggg agtacttcat ccctaaagag gccaaagaga tcgactacca
     6901 caagcactac gccagagcct tcgacaagta caccaacctg cagaaagaag gcatcgacgc
     6961 catcctgaaa ctggaaggca gaatcatcga gcgccggaag atcaagcccg ccaagaacta
     7021 catcgagttt caagagatta tgaatcggag cggctacaac aacgaccagc aggttgccct
     7081 gaagagagtg gcgaacgccc tgctggccta caacctgaac ttcgagagag agcacctgaa
     7141 gcggttctac ggcgtcgtga agagagaggg catcgagaaa aaatggtccc tgatcgtggg
     7201 agggtcaggc ggtagtcagc tgcatttacc gcaggtttta gctgacgctg tctcacgcct
     7261 ggtcctgggt aagtttggtg acctgaccga caacttctcc tcccctcacg ctcgcagaaa
     7321 agtgctggct ggagtcgtca tgacaacagg cacagatgtt aaagatgcca aggtgataag
     7381 tgtttctaca ggaacaaaat gtattaatgg tgaatacatg agtgatcgtg gccttgcatt
     7441 aaatgactgc catgcagaaa taatatctcg gagatccttg ctcagatttc tttatacaca
     7501 acttgagctt tacttaaata acaaagatga tcaaaaaaga tccatctttc agaaatcaga
     7561 gcgagggggg tttaggctga aggagaatgt ccagtttcat ctgtacatca gcacctctcc
     7621 ctgtggagat gccagaatct tctcaccaca tgagccaatc ctggaagaac cagcagatag
     7681 acacccaaat cgtaaagcaa gaggacagct acggaccaaa atagagtctg gtcaggggac
     7741 gattccagtg cgctccaatg cgagcatcca aacgtgggac ggggtgctgc aaggggagcg
     7801 gctgctcacc atgtcctgca gtgacaagat tgcacgctgg aacgtggtgg gcatccaggg
     7861 atcactgctc agcattttcg tggagcccat ttacttctcg agcatcatcc tgggcagcct
     7921 ttaccacggg gaccaccttt ccagggccat gtaccagcgg atctccaaca tagaggacct
     7981 gccacctctc tacaccctca acaagccttt gctcagtggc atcagcaatg cagaagcacg
     8041 gcagccaggg aaggccccca acttcagtgt caactggacg gtaggcgact ccgctattga
     8101 ggtcatcaac gccacgactg ggaaggatga gctgggccgc gcgtcccgcc tgtgtaagca
     8161 cgcgttgtac tgtcgctgga tgcgtgtgca cggcaaggtt ccctcccact tactacgctc
     8221 caagattacc aagcccaacg tgtaccatga gtccaagctg gcggcaaagg agtaccaggc
     8281 cgccaaggcg cgtctgttca cagccttcat caaggcgggg ctgggggcct gggtggagaa
     8341 gcccaccgag caggaccagt tctcactcac gtaagcggcc gctcgagtct agagggcccg
     8401 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     8461 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     8521 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     8581 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     8641 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     8701 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     8761 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     8821 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     8881 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     8941 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     9001 tgttccaaa
//
