LOCUS       sequence-437823-        4148 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Effector only expresses Cas7.1 domain (truncated from DiCas7-11 
            effector), nuclear localization signal sequence on 5' and 3' end of 
            the coding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4148)
  TITLE     CRISPR-Cas transgene repressible element
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4148)
  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            "Unpublished"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: 1203T-CMV-Cas7.1-NLS-bGHpolyA
FEATURES             Location/Qualifiers
     source          1..4148
                     /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(2092..2112)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(2160..2363)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2189..2209)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2364..2743
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2915..2934
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3013..3117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3118..3186,3187..3978)
                     /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(3336..3355)
                     /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 gtccgaacct
     1381 gattacgcac agagctccac acagccgatc cgtaaacttg agagaatcac agagcagttt
     1441 cctggcttca gcgctgactt tattatctat ggtaatctcc ccttcgaagc ggggaaacct
     1501 agtatggtcg acttcgtaag ccttaaagaa gtcatgggcc ttaccggatt tgaaatcgac
     1561 cctgttaagt acccgttggc tgcctatagc ttttggcccg tcgaaatcgg gcttccctgg
     1621 aagggaaaga ttgccaaaat gaatacgaaa gcgccaatcc ttcttctcag ccttctttcc
     1681 atcgttaccg gatctcccga gaagttcgca aaacgggcag taagtttcag cgttatctgg
     1741 acaggggttc ttatcggtcc aacgaagggt ggaccgctgc ctcaattgca gcaacctgtc
     1801 tttatcttcc gtatcaaatt tcccaggaca acatgtcttt tcgctaattt tcccatcact
     1861 caatgtcagc aggttttccg cgcttctaat tactgcggac cgcagcaatg tcccagttat
     1921 gtaaggccgc ccttttgtgt tatccttctt atttctatgc catcttgcaa aagcttggcc
     1981 gcgaacaaat tccttgttgt tcttgtttcg cctagtgctt tcctgccatt tggtcatccg
     2041 aaacggctcg aggaattcaa ttgaaatctt catagtagtc gtgctggcct ccacctttct
     2101 cttcttcttg gggctcatgc ggccgcggat ctctagcgga tctgacggtt cactaaacca
     2161 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
     2221 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     2281 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     2341 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     2401 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     2461 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     2521 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     2581 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     2641 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     2701 cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt atatctggcc
     2761 cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat gcaattgtcg
     2821 gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc gacctccaac
     2881 acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     2941 gtcgacccta ggggatcggg agatcgatct cccgatccgt cgacgtcagg tggcactttt
     3001 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3061 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3121 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3181 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3241 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3301 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3361 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3421 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3481 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3541 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3601 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3661 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3721 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3781 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3841 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3901 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3961 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4021 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4081 aaaatccctt aacgtgagtt ttcgttccac tgagtgtcag accccgtaga aaagatcaaa
     4141 ggatcttc
//
