LOCUS       sequence-418032-        7127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SynCas.v39 for analysis of RNA knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7127)
  AUTHORS   Brogan DJ, Dalla Benetta E, Wang T, Lin CP, Chen F, Li H, Lin C, 
            Komives EA, Akbari OS
  TITLE     Synthetic type III-E CRISPR-Cas effectors for programmable 
            RNA-targeting
  JOURNAL   bioRxiv 2024.02.23.581838; doi: 
            https://doi.org/10.1101/2024.02.23.581838
REFERENCE   2  (bases 1 to 7127)
  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: "bioRxiv 
            2024.02.23.581838; doi: https://doi.org/10.1101/2024.02.23.581838"
COMMENT     Sequence Label: CMV-SynCas.v39 mammalian expression
FEATURES             Location/Qualifiers
     source          1..7127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     532..551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(770..874)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(953..972)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1144..1523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1524..1727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1678..1698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5534..5551
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5574..5591)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5580..5804
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5875..5891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5875..5891)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5888..5910)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5899..5915
                     /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(5923..5929,5930..5947,5948..5953))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5968..5989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6106..6123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6277..6865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6357..6376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7036..7127,1..700,701..769))
                     /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"
ORIGIN
        1 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
       61 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      121 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
      181 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
      241 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      301 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
      361 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
      421 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
      481 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
      541 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
      601 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
      661 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
      721 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
      781 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
      841 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
      901 acgtcgacgg atcgggagat cgatctcccg atcccctagg gtcgactctc agtacaatct
      961 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     1021 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     1081 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     1141 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     1201 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     1261 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     1321 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     1381 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     1441 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     1501 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     1561 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     1621 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     1681 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     1741 gtcagatccg ctagagatcc gcggccgcat gacgactact atgaagattt caattgaatt
     1801 cctcgagccg tttcggatga ccaaatggca ggaaagcact aggcgaaaca agaacaacaa
     1861 ggaatttgtt cgcggccaag cttttgcaag atggcataga aataagaagg ataacacaaa
     1921 agggcggcct tacataactg ggacattgct gcggtccgca gtaattagaa gcgcggaaaa
     1981 cctgctgaca ttgagtgatg ggaaaattag cgaaaagaca tgttgtcctg ggaaatttga
     2041 tacggaagat aaagacaggt tgctgcaatt gaggcagcgg tccacccttc gttggaccga
     2101 taagaacccc tgtccagata acgctgaaac ttactgcccg ttttgcgaac ttctcgggag
     2161 atccggtaac gatggaaaga aggctgagaa gaaggattgg cgctttcgta ttcattttgg
     2221 caatctttcc cttccaggga agcccgattt cgacgggcca aaagctatag gcagccaacg
     2281 ggtacttaac agggtcgatt tcaaatccgg taaggcccat gacttcttta aggcttacga
     2341 agtcgaccat actaggtttc cccgcttcga aggggagatt accatagata ataaagtcag
     2401 cgctgaagcc aggaaactgc tctgtgattc tctcaagttt acggatcggc tgtgtggagc
     2461 tctgtgcgta atcaggttcg atgaatatac accagcagcc gatagtggga aacaaaccga
     2521 aaatgtccaa gcagaaccga atgctaatct cgctgaaaag accgcagagc aaattattag
     2581 cattctggac gataataaga aaaccgaata tacccgcctg cttgctgatg ctattcgttc
     2641 actccgccgc tctagtaaac ttgtcgctgg cttgccaaag gatcatgacg ggaaggatga
     2701 ccattacctc tgggatatcg gcaagaagaa gaaagacgaa aactctgtca ctattaggca
     2761 aatccttacg acctcagcag acaccaagga actcaagaat gccggaaaat ggagagaatt
     2821 ctgcgagaag ctgggtgaag cgctgtacct caagagtaaa gacatgagtg gcggcctgaa
     2881 aattactcgg agaatactgg gcgatgctga attccatgga aagcccgatc ggcttgaaaa
     2941 gagccggtct gtgtcaatcg gatctgtgtt gaaagaaact gtggtatgcg gcgaactggt
     3001 cgctaaaacg cccttcttct tcggagcgat agacgaagat gcaaaacaaa ccgacctgca
     3061 agtactcctc actcccgata acaagtatag actgccaaga agcgccgtgc gaggtatact
     3121 ccgtcgagat cttcaaacct attttgatag cccatgcaat gctgagttgg gtggacggcc
     3181 atgcatgtgt aagacgtgta ggattatgag agggatcacg gtgatggatg cgcgcagtga
     3241 gtacaatgcc ccgccagaaa taaggcatcg tacccgcatt aatcccttca caggcacggt
     3301 tgcggaaggt gccctgttta atatggaagt agcccccgag gggattgtct ttccatttca
     3361 actccggtac cggggctctg aagatgggct gcccgatgca ctgaaaacgg tgttgaaatg
     3421 gtgggctgag gggcaggcat tcatgagtgg cgctgcctca accgggaagg gccgattccg
     3481 gatggaaaat gctaaatatg aaacgctgga tctgagcgac gagaatcaaa ggaatgacta
     3541 tcttaagaat tggggatggc gtgacgagaa ggggctcgag gaactgaaga aacgactgaa
     3601 ctcaggtctg ccagagcccg gtaattatag ggatccaaaa tggacaaaag tagggtatga
     3661 gattaaaata cagtcgcccc ttcttagcaa cgatcctatc gcagcattgt tcgacactgc
     3721 aaaccacgac gcggtcgcgt ataagaaacg agtgtgggag aacggacgta tcatagagcg
     3781 acacactttc aaaggagagt ctatacgagg ggtgctgcgc actgcacttg ggcgtgagta
     3841 cggattgttc gaattaaaac acgaagactg tccttgtagt ctttgcacaa ttttcggtaa
     3901 cgaacacgaa gcgggcaaaa tacgtttcga agacttagaa atcgaggacg gtgttgccga
     3961 aaagcacttc gatcacgttg ctatcgatcg cttcacaggc ggagccgtag acaagaagaa
     4021 attcgacgat ttctctttag ctggttcccc gggaaagccc attaaactca aagggtgctt
     4081 ttggttgcgt gcggatatat ttagtgacga gaagaaggac gagaagaaga aaataataaa
     4141 cgctctttgt gacgtgcgtg atggcttgta ccctctcggt ggcaagggcg gtgtcggata
     4201 cgggtgggta aaagagctga gaataacgga cgatgataag aggatttctc gcctcatgaa
     4261 tcctgccttt gatgagacag atgtagccgt tccagaaaag cccaaaactg atgccgaggt
     4321 tcgcatcgag gcagagaaag tatattaccc acactatttc gtcgaacccc ataagaaggt
     4381 ggaacgcgag gagaaaccct gtggtcatca aaagttccac gaggggcgac tgactggtaa
     4441 aattgagtgt acactgaaaa cgctgagccc tttaataata ccggacagtg aggtcgttac
     4501 ggagaacgga aacggccaca agtcttacca gttcttcaga ttaaataatg aaatcatcat
     4561 tccgggaagc gaaattcggg gtatgatcag ttccgtgtac gaagcactta ctaactcttg
     4621 cttccgtgtc gtggacgaca tcatcccggt ggtgatctcc cgtagcgtgt caccgtacca
     4681 gatcgctcag aagctgccca gtccggatct gcgtccttgc gagcgtgaga ttcttgagga
     4741 ctcagaaatt ggatctttag acaagtgttc agagaaacgc cttttccgcc tgcatgaaga
     4801 cggtctttgc ccatcgtgtc gtctgttcgg cactacacat tataagggac gcgttcggtt
     4861 cggcttcgca aagcacgagg gcggcgaaaa gtggctcatg gacgacaaga ctaaggacga
     4921 gtatctgact ttaccgttgt tggaacggcc gcgcccaaca tggagcatgc caagcgacga
     4981 agacgacgta ccgggacgga aattctacgt acaccacaat ggttgggaga aagttaagca
     5041 agatagttta gataataaaa cgagcaaaac cgagaataat cgaagcgtgg aaccactcga
     5101 taaggataac ctgtttaaat tcgacgtgtt cttcgagaac ctggaaccct gggaacttgg
     5161 tttgcttctc tactcattag aattggaaga ggggttagcc cacaagtttg gaatggcaaa
     5221 agcgttcggc ttcggctcga caaagattga cgccgacaaa atactgttac gtgaagaggc
     5281 cgggaagttc cacccttgcg agaaagcaga cgaatacctt aagaagggct tgaacaagct
     5341 gaccagctgg ttcggtaaga acgaaatcaa tgagcacatg cgaaacttac tcttgttcat
     5401 gacttactat cagaaccttc ccaaggttaa gtaccctgat tttgacggat acgcaaaatg
     5461 gcggtgcagc tacgttgaac aggacaaggt ggaatacttt cagaaccgct ggattgtggc
     5521 ctcttaaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     5581 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     5641 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     5701 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     5761 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     5821 aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5881 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5941 cataaagtgt aaagcctagg atgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6001 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6061 acgcgcggga agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     6721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacac
     6901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataacta
//
