LOCUS       sequence-437816-        4082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ffector expresses Cas7.1 domain (truncated from DiCas7-11), N to A 
            mutation at amino acid 152.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4082)
  TITLE     CRISPR-Cas transgene repressible element
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4082)
  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            "Unpublished"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: 1203AD-CMV-Cas7.1-N152A-mutant
FEATURES             Location/Qualifiers
     source          1..4082
                     /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(2094..2297)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2123..2143)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2298..2677
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2849..2868
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2947..3051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3052..3120,3121..3912)
                     /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(3270..3289)
                     /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 agaacctgat tacgcacaga gctccacaca gccgatccgt
     1381 aaacttgaga gaatcacaga gcagtttcct ggcttcagcg ctgactttat tatctatggt
     1441 aatctcccct tcgaagcggg gaaacctagt atggtcgact tcgtaagcct taaagaagtc
     1501 atgggcctta ccggatttga aatcgaccct gttaagtacc cgttggctgc ctatagcttt
     1561 tggcccgtcg aaatcgggct tccctggaag ggaaagagcg ccaaaatgaa tacgaaagcg
     1621 ccaatccttc ttctcagcct tctttccatc gttaccggat ctcccgagaa gttcgcaaaa
     1681 cgggcagtaa gtttcagcgt tatctggaca ggggttctta tcggtccaac gaagggtgga
     1741 ccgctgcctc aattgcagca acctgtcttt atcttccgta tcaaatttcc caggacaaca
     1801 tgtcttttcg ctaattttcc catcactcaa tgtcagcagg ttttccgcgc ttctaattac
     1861 tgcggaccgc agcaatgtcc cagttatgta aggccgccct tttgtgttat ccttcttatt
     1921 tctatgccat cttgcaaaag cttggccgcg aacaaattcc ttgttgttct tgtttcgcct
     1981 agtgctttcc tgccatttgg tcatccgaaa cggctcgagg aattcaattg aaatcttcat
     2041 agtagtcgtc atgcggccgc ggatctctag cggatctgac ggttcactaa accagctctg
     2101 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     2161 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     2221 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     2281 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     2341 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     2401 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     2461 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     2521 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     2581 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     2641 attgattact attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca
     2701 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag
     2761 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa
     2821 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtcgac
     2881 cctaggggat cgggagatcg atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     2941 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3001 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3061 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3121 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3181 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3241 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3301 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3361 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3421 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3481 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3541 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3601 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3661 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3721 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3781 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3841 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3901 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     3961 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4021 ccttaacgtg agttttcgtt ccactgagtg tcagaccccg tagaaaagat caaaggatct
     4081 tc
//
