LOCUS       sequence-291991-        5733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive PGK driven expression of N terminal dCpf1 (Cas12a) 
            406-split half with leucine zipper.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5733)
  AUTHORS   Kempton HR, Goudy LE, Love KS, Qi LS
  TITLE     Multiple Input Sensing and Signal Integration Using a Split Cas12a 
            System.
  JOURNAL   Mol Cell. 2020 Jan 30. pii: S1097-2765(20)30037-X. doi: 
            10.1016/j.molcel.2020.01.016.
  PUBMED    32027839
REFERENCE   2  (bases 1 to 5733)
  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: "Mol Cell. 
            2020 Jan 30. pii: S1097-2765(20)30037-X. doi: 
            10.1016/j.molcel.2020.01.016."
COMMENT     Sequence Label: pSLQ8518 PB-PGK-N term (1-406) dLbCpf1-EE12RR3...
FEATURES             Location/Qualifiers
     source          1..5733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..500
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(36..59)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     433..452
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        complement(1936..1954)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1937..1954)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1974..1991)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1980..2204
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2420..2482
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2533..2553)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2533..2551)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2572..2588)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2572..2588)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2585..2607)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2596..2612
                     /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(2620..2626,2627..2644,2645..2650))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2665..2686
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2803..2820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2974..3562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3054..3073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3733..4524,4525..4593))
                     /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     4356..4375
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4594..4698)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4724..5179)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4861..4882)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5073..5092
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5306..5328
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5320..5337
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5321..5337
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5347..5366
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5347..5365
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5395..5429
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
       61 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
      121 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
      181 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
      241 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
      301 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
      361 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
      421 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
      481 cctcatctcc gggcctttcg agaattcggt accatgagca agctggagaa gtttacaaac
      541 tgctactccc tgtctaagac cctgaggttc aaggccatcc ctgtgggcaa gacccaggag
      601 aacatcgaca ataagcggct gctggtggag gacgagaaga gagccgagga ttataagggc
      661 gtgaagaagc tgctggatcg ctactatctg tcttttatca acgacgtgct gcacagcatc
      721 aagctgaaga atctgaacaa ttacatcagc ctgttccgga agaaaaccag aaccgagaag
      781 gagaataagg agctggagaa cctggagatc aatctgcgga aggagatcgc caaggccttc
      841 aagggcaacg agggctacaa gtccctgttt aagaaggata tcatcgagac aatcctgcca
      901 gagttcctgg acgataagga cgagatcgcc ctggtgaaca gcttcaatgg ctttaccaca
      961 gccttcaccg gcttctttga taacagagag aatatgtttt ccgaggaggc caagagcaca
     1021 tccatcgcct tcaggtgtat caacgagaat ctgacccgct acatctctaa tatggacatc
     1081 ttcgagaagg tggacgccat ctttgataag cacgaggtgc aggagatcaa ggagaagatc
     1141 ctgaacagcg actatgatgt ggaggatttc tttgagggcg agttctttaa ctttgtgctg
     1201 acacaggagg gcatcgacgt gtataacgcc atcatcggcg gcttcgtgac cgagagcggc
     1261 gagaagatca agggcctgaa cgagtacatc aacctgtata atcagaaaac caagcagaag
     1321 ctgcctaagt ttaagccact gtataagcag gtgctgagcg atcgggagtc tctgagcttc
     1381 tacggcgagg gctatacatc cgatgaggag gtgctggagg tgtttagaaa caccctgaac
     1441 aagaacagcg agatcttcag ctccatcaag aagctggaga agctgttcaa gaattttgac
     1501 gagtactcta gcgccggcat ctttgtgaag aacggccccg ccatcagcac aatctccaag
     1561 gatatcttcg gcgagtggaa cgtgatccgg gacaagtgga atgccgagta tgacgatatc
     1621 cacctgaaga agaaggccgt ggtgaccgag aagtacgagg acgatcggag aaagtccttc
     1681 aagaagatcg gctccttttc tctggagcag ctgcaggagt acgccgacgc caagcttgga
     1741 ggaggtggtg gaggtggagg tggaggtact agtctggaga tcgaggccgc cttcctggag
     1801 cgcgagaaca ccgccctgga gacccgcgtg gccgagctgc gccagcgcgt gcagcgcctg
     1861 cgcaaccgcg tgagccagaa ccgcacccgc tacggccccc tgggcggcgg caagtaactc
     1921 gagagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     1981 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2041 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2101 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2161 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggtcgaggc
     2221 ggccgctgat cacaattgtc tagttaaaag ttttgttact ttatagaaga aattttgagt
     2281 ttttgttttt ttttaataaa taaataaaca taaataaatt gtttgttgaa tttattatta
     2341 gtatgtaagt gtaaatataa taaaacttaa tatctattca aattaataaa taaacctcga
     2401 tatacagacc gataaaacac atgcgtcaat tttacgcatg attatcttta acgtacgtca
     2461 caatatgatt atctttctag ggttaatcta gtatacgcgt atgcggccgc ttaattaatc
     2521 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     2581 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2641 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2701 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2761 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2821 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2881 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2941 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3001 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3061 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3121 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3181 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3241 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3301 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3361 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3421 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3481 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3541 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3601 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3661 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3721 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3781 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3841 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3901 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3961 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4021 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4081 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4141 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4201 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4261 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4321 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4381 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4441 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4501 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4561 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4621 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4681 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     4741 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4801 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4861 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4921 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4981 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5041 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5101 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5161 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5221 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     5281 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     5341 cgcgcgtaat acgactcact atagggcgaa ttggggcgcg ccattctaga ttaaccctag
     5401 aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc tctttctaaa
     5461 tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga gctcccgtga
     5521 ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc gcgtgagtca
     5581 aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg ataattatat
     5641 tgttatttca tgttctactt acgtgataac ttattatata tatattttct tgttatagat
     5701 atcaactaga atgctagcat gggcccagat ctc
//
