LOCUS       sequence-437448-        8246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Deactivated DiCas7-11 effector with CMV promoter/enhancer and 
            nuclear localization signal sequence at 5' and 3' end of the coding 
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8246)
  TITLE     CRISPR-Cas transgene repressible element
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8246)
  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            "Unpublished"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: 1203N-CMV-dDiCas7-11-NLS-bGHpolyA
FEATURES             Location/Qualifiers
     source          1..8246
                     /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"
     CDS             complement(1351..1371)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6190..6210)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(6258..6461)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6287..6307)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6462..6841
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     7013..7032
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7111..7215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7216..7284,7285..8076)
                     /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(7434..7453)
                     /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 aagcggccgc caccttcctc tttttcttag gtccggatcc
     1381 tgcccacggt gtccacggcg tggtcagctt cttctgccga tcttctttct tgaattcccc
     1441 gtctttgagt tcttcgtaac cgctattccc attaggatcg tccttctttc gcagcatggg
     1501 gtagcaaacc cggggtgctt ggtcgccctc aggaaaccaa agaagtttct taagattttc
     1561 tataaaatcg agctcgtccc ggaaccattc cttcagttta gcaaatcctt ttcccaacca
     1621 gttgggaatt tcggaattgc cattccgcca ttgcttccaa tcttttctca ggcgcactga
     1681 ttcgacgtca atttcaactg aaccaaatcc catagacttt gccattccaa gtttatgcgc
     1741 cagacccttt tcgagttgca ggctgtggat gagcaaaccc agctcccatt ctttcaggtt
     1801 ctcaaaggca atttcgaagc taaaggaatt cccgccggcg agagcttcta cagtgcggtt
     1861 attagggctt tgttcaatag ctttgcctgt tgtgggatgg ttcccgtcct taatggtttt
     1921 ccaggcgtgg tggtggacgt aaaactttct gccaggcact ttaaacttat tgtcggatcc
     1981 cggtatactc caagtcgggc gaggccgttc caacaatgag agcatcaccg gcccgccgtg
     2041 aaacggatcg ccgggattct tgcctgggat cagccactcc ggatcgtttt cgagtgacgc
     2101 gaatccaaac ctgactctgc ccttgtaaga tccggttcca aagagccggc aagcagggca
     2161 gagtcctttg gggtggcgca agaggagtct tttctcggga tatgcattca aggcactgag
     2221 atcaccatct tccacccaat ctccatggca cggccgcaga tcggcgctca tacgtttgcc
     2281 gatcatccta tcgtctacag ttcgggaaat ccgcactggg acgatgtcct ctacgtactt
     2341 ttcattatgt ttgaaataca caagatctcc ggacttcagc ttttccacgt tttcccgggc
     2401 aactctagac tgaaatacgc tctccggcac tgcttgagga ttgttgttat agaccctgag
     2461 cagttctttg tatttgattc ttgctttctc ggggatctcg tattcctcgt tctccttcag
     2521 atcgaagaag aaggtttcgc aatatttagc catggtatag taattgccct tatcatcctc
     2581 acaacaaaac acaggttcat tcttcctttt cctattctta aaatcgttcg tcctaattgt
     2641 tttccagtcg ttgggaactg atgggagtgt tccttgaaag ttattgatca catccccttt
     2701 cttgtcacta aactctactt tgctggggcc gacgacatga aggtagccct ctttacactc
     2761 ccatccgtct gggtcgttct cgggtatctc ttgaaaggcc ttttggtatc tactatctct
     2821 tacccagcaa tccagtttct caccattctg tatgccttcg agtttctcct ggtcaaaatt
     2881 gtcaggttta tgccacaagt ggaaagaagt gcagccatta ttggtgacca cgttaaaata
     2941 ataagagcca ttcttttgct ctataaatgt ggcgatgcct tctttgcgtc ttttccattt
     3001 attgtcttgt ttcttctgtg ggtgtttagc aggatcaacg aggctcagtc gtttaataaa
     3061 acgcttgacc cacatccgaa caggtttctc accagcaatt tcttgctcat caagaatgtc
     3121 aaaatgtttc tgcgttttgt catagaaagg cacccttgca gtttcggaaa acttttgtat
     3181 atgtttccca tcggcggtca ctcgacctgg gagaaagtct tgcagcacat tctggtgatc
     3241 agcatccatc cgccatgaca accgtttagt ctcatcaaat atcctgaagc aagaatttgt
     3301 cacagtttcg tagacactac tcaccattcc tctcaattcg ctcccgggaa tcatgatttg
     3361 tttatgaagt ctgaagaaag catatgattt atgatattcg tccttttcct tccgagcttc
     3421 tttatccgca ggtctgaaga aatcgtcatt actagtatct gggacgatga ggggtgtctt
     3481 ggtaatgagc ttacaccgaa ttttacctgt cagtcgcccc tcgtggaact tttgatgacc
     3541 acagggtttc tcctcgcgtt ccaccttctt atggggttcg acgaaatagt gtgggtaata
     3601 tactttctct gcctcgatgc gaacctcggc atcagttttg ggcttttctg gaacggctac
     3661 atctgtctca tcaaaggcag gattcatgag gcgagaaatc ctcttatcat ctccctttat
     3721 acccagactc ttgacctgtc cgtagccgat ggcgctcttt ccaccgaggg gataaaggcc
     3781 attattcaca tctgcaagag ccttaccgag tgccttacag tactcttcgt cctcgagaac
     3841 gtccctgcgg atccagaagg agcctttcaa catgagcggt cttgcagggc taccgggcag
     3901 aggggaatca tcgaatttct tcttatcagc tgcaccgcca gtgaatctat cgatggccac
     3961 atgatcgaag gtcacaggtt ccgggtcact ttcgaatacg agatcttcaa accttatttt
     4021 gccggcttca tactcacttc caaagatttg gcacaggaga cattcacagt cactgtgagt
     4081 caactcagtc aaagggacgc catcttccat atgaatgcgt gccacggcac ttctgatcac
     4141 ccctctaaaa gattcggcct tgtatgcaca tacgggtttc gcctcttccc cttcagcttt
     4201 atacttaacg aaggtaacca ccgctgtacc acgtttgtcc acggctgcgc ggattgggtc
     4261 gccattaata aaagggcttg ccatctcaat ggaaacgtta atctcgtgcc attttggatc
     4321 cctataatta ccgggctctg gcagacctga gttcagtcgt ttcttcagtt cctcgagccc
     4381 cttctcgtca cgccatcccc aattcttaag atagtcattc ctttgattct cgtcgctcag
     4441 atccagcgtt tcatatttag cattttccat ccggaatcgg cccttcccgg ttgaggcagc
     4501 gccactcatg aatgcctgcc cctcagccca ccatttcaac accgttttca gtgcatcggg
     4561 cagcccatct tcagagcccc ggtaccggag ttgaaatgga aagacaatcc cctcgggggc
     4621 tacttccata ttaaacaggg caccttccgc aaccgtgcct gtgaagggat taatgcgggt
     4681 acgatgcctt atttctggcg gggcattgta ctcactgcgc gcatccatca ccgtgatccc
     4741 tctcataatc ctacacgtct tacacatgca tggccgtcca cccaactcag cattgcatgg
     4801 gctatcaaaa taggtttgaa gatctcgacg gagtatacct cgcacggcgc ttcttggcag
     4861 tctatacttg ttatcgggag tgaggagtac ttgcagtgcg gtttgttttg catcttcgtc
     4921 tatcgctccg aagaagaagg gcgttttagc gaccagttcg ccgcatacca cagtttcttt
     4981 caacacagat ccgattgaca cagaccggct cttttcaagc cgatcgggct ttccatggaa
     5041 ttcagcatcg cccagtattc tccgagtaat tttcaggccg ccactcatgt ctttactctt
     5101 gaggtacagc gcttcaccca gcttctcgca gaattctctc cattttccgg cattcttgag
     5161 ttccttggtg tctgctgagg tcgtaaggat ttgcctaata gtgacagagt tttcgtcttt
     5221 cttcttcttg ccgatatccc agaggtaatg gtcatccttc ccgtcatgat cctttggcaa
     5281 gccagcgaca agtttactag agcggcggag tgaacgaata gcatcagcaa gcaggcgggt
     5341 atattcggtt ttcttattat cgtccagaat gctaataatt tgctctgcgg tcttttcagc
     5401 gagattagca ttcggttctg cttggacatt ttcggtttgt ttcccactat cggctgctgg
     5461 tgtatattca tcgaacctga ttacgcacag agctccacac agccgatccg taaacttgag
     5521 agaatcacag agcagtttcc tggcttcagc gctgacttta ttatctatgg taatctcccc
     5581 ttcgaagcgg ggaaacctag tatggtcgac ttcgtaagcc ttaaagaagt catgggcctt
     5641 accggatttg aaatcgaccc tgttaagtac ccgttggctg cctatagctt ttggcccgtc
     5701 gaaatcgggc ttccctggaa gggaaagatt gccaaaatga atacgaaagc gccaatcctt
     5761 cttctcagcc ttctttccat cgttaccgga tctcccgaga agttcgcaaa acgggcagta
     5821 agtttcagcg ttatctggac aggggttctt atcggtccaa cgaagggtgg accgctgcct
     5881 caattgcagc aacctgtctt tatcttccgt atcaaatttc ccaggacaac atgtcttttc
     5941 gctaattttc ccatcactca atgtcagcag gttttccgcg cttctaatta ctgcggaccg
     6001 cagcaatgtc ccagttatgt aaggccgccc ttttgtgtta tccttcttat ttctatgcca
     6061 tcttgcaaaa gcttggccgc gaacaaattc cttgttgttc ttgtttcgcc tagtgctttc
     6121 ctgccatttg gtcatccgaa acggctcgag gaattcaatt gaaatcttca tagtagtcgt
     6181 gctggcctcc acctttctct tcttcttggg gctcatgcgg ccgcggatct ctagcggatc
     6241 tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc
     6301 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
     6361 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
     6421 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
     6481 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     6541 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     6601 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     6661 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
     6721 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
     6781 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt
     6841 caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga
     6901 ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac
     6961 tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca
     7021 gagcagattg tactgagagt cgaccctagg ggatcgggag atcgatctcc cgatccgtcg
     7081 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     7141 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     7201 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     7261 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     7321 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     7381 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     7441 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7501 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7561 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7621 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7681 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7741 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7801 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7861 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7921 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7981 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     8041 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     8101 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     8161 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agtgtcagac
     8221 cccgtagaaa agatcaaagg atcttc
//
