LOCUS       sequence-437431-        5114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Effector expresses Cas7.1, Linker1, Cas11, Linker 2, and deactivated
            Cas7.2 domains from DiCas7-11, H to A mutation at amino acid 43 in 
            Cas7.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5114)
  TITLE     CRISPR-Cas transgene repressible element
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5114)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 1203AE-CMV-Cas7.1-7.2-H43A-mutant
FEATURES             Location/Qualifiers
     source          1..5114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1062..1286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1275..1292
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1315..1332)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(3126..3329)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3155..3175)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3330..3709
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3881..3900
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3979..4083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4084..4152,4153..4944)
                     /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     complement(4302..4321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctcttcccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcatcctag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtatc gagccccagc tggttctttc cgcctcagaa gccatagagc ccaccgcatc
     1081 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
     1141 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
     1201 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
     1261 agatggctgg caactagaag gcacagtcga ggctgatcag cgggtttaaa ctcaatggtg
     1321 atggtgatga tgaccggttt aatccagcgt ttcatattta gcattttcca tccggaatcg
     1381 gcccttcccg gttgaggcag cgccactcat gaatgcctgc ccctcagccc accatttcaa
     1441 caccgttttc agtgcatcgg gcagcccatc ttcagagccc cggtaccgga gttgaaatgg
     1501 aaagacaatc ccctcggggg ctacttccat attaaacagg gcaccttccg caaccgtgcc
     1561 tgtgaaggga ttaatgcggg tacgatgcct tatttctggc ggggcattgt actcactgcg
     1621 cgcatccatc accgtgatcc ctctcataat cctacacgtc ttacacatgc atggccgtcc
     1681 acccaactca gcattgcatg ggctatcaaa ataggtttga agatctcgac ggagtatacc
     1741 tcgcacggcg cttcttggca gtctatactt gttatcggga gtgaggagta cttgcagtgc
     1801 ggtttgtttt gcatcttcgt ctatcgctcc gaagaagaag ggcgttttag cgaccagttc
     1861 gccgcatacc acagtttctt tcaacacaga tccgattgac acagaccggc tcttttcaag
     1921 ccgatcgggc tttccatgga attcagcatc gcccagtatt ctccgagtaa ttttcaggcc
     1981 gccactcatg tctttactct tgaggtacag cgcttcaccc agcttctcgc agaattctct
     2041 ccattttccg gcattcttga gttccttggt gtctgctgag gtcgtaagga tttgcctaat
     2101 agtgacagag ttttcgtctt tcttcttctt gccgatatcc cagaggtaat ggtcatcctt
     2161 cccgtcatga tcctttggca agccagcgac aagtttacta gagcggcgga gtgaacgaat
     2221 agcatcagca agcaggcggg tatattcggt tttcttatta tcgtccagaa tgctaataat
     2281 ttgctctgcg gtcttttcag cgagattagc attcggttct gcttggacat tttcggtttg
     2341 tttcccacta tcggctgctg gtgtatattc atcgaacctg attacgcaca gagctccaca
     2401 cagccgatcc gtaaacttga gagaatcaca gagcagtttc ctggcttcag cgctgacttt
     2461 attatctatg gtaatctccc cttcgaagcg gggaaaccta gtatggtcga cttcgtaagc
     2521 cttaaagaag tcatgggcct taccggattt gaaatcgacc ctgttaagta cccgttggct
     2581 gcctatagct tttggcccgt cgaaatcggg cttccctgga agggaaagat tgccaaaatg
     2641 aatacgaaag cgccaatcct tcttctcagc cttctttcca tcgttaccgg atctcccgag
     2701 aagttcgcaa aacgggcagt aagtttcagc gttatctgga caggggttct tatcggtcca
     2761 acgaagggtg gaccgctgcc tcaattgcag caacctgtct ttatcttccg tatcaaattt
     2821 cccaggacaa catgtctttt cgctaatttt cccatcactc aatgtcagca ggttttccgc
     2881 gcttctaatt actgcggacc gcagcaatgt cccagttatg taaggccgcc cttttgtgtt
     2941 atccttctta tttctagccc atcttgcaaa agcttggccg cgaacaaatt ccttgttgtt
     3001 cttgtttcgc ctagtgcttt cctgccattt ggtcatccga aacggctcga ggaattcaat
     3061 tgaaatcttc atagtagtcg tcatgcggcc gcggatctct agcggatctg acggttcact
     3121 aaaccagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     3181 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     3241 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     3301 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     3361 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     3421 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     3481 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     3541 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     3601 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     3661 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat
     3721 ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa
     3781 ttgtcggtca agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc
     3841 tccaacacac aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta
     3901 ctgagagtcg accctagggg atcgggagat cgatctcccg atccgtcgac gtcaggtggc
     3961 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4321 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag tgtcagaccc cgtagaaaag
     5101 atcaaaggat cttc
//
