LOCUS       sequence-292003-        7332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive PGK driven expression of leucine zipper with C terminal
            dCpf1 (Cas12a) 406-split half fused to intein for 3-way split 
            activator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7332)
  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 7332)
  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: pSLQ8636 PB-PGK-RR12EE345L-C term (406) dLbCpf...
FEATURES             Location/Qualifiers
     source          1..7332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..529
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(65..88)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     462..481
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3201..3227
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3207..3233
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(3564..3582)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3565..3582)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3602..3619)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3608..3832
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4048..4110
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4161..4181)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4161..4179)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4200..4216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4200..4216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4213..4235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4224..4240
                     /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(4248..4254,4255..4272,4273..4278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4293..4314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4431..4448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4602..5190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4682..4701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5361..6152,6153..6221))
                     /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     5984..6003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6222..6326)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6352..6807)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6489..6510)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6701..6720
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6934..6956
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6948..6965
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6949..6965
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6975..6994
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6975..6993
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   7023..7057
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 actagaatgc tagcatgggc ccagatctcg ggtaggggag gcgcttttcc caaggcagtc
       61 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
      121 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
      181 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
      241 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
      301 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
      361 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
      421 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
      481 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga gaattcggta
      541 ccatgctgga gatccgcgcc gccttcctgc gccagcgcaa caccgccctg cgcaccgagg
      601 tggccgagct ggagcaggag gtgcagcgcc tggagaacga ggtgagccag tacgagaccc
      661 gctacggccc cctgggcggc ggcaaggcta gcggaggagg tggtggaggt ggaggtggag
      721 gtcatatgga tctgtctgtg gtggagaagc tgaaggagat catcatccag aaggtggatg
      781 agatctacaa ggtgtatggc tcctctgaga agctgttcga cgccgatttt gtgctggaga
      841 agagcctgaa gaagaacgac gccgtggtgg ccatcatgaa ggacctgctg gattctgtga
      901 agagcttcga gaattacatc aaggccttct ttggcgaggg caaggagaca aacagggacg
      961 agtccttcta tggcgatttt gtgctggcct acgacatcct gctgaaggtg gaccacatct
     1021 acgatgccat ccgcaattat gtgacccaga agccctactc taaggataag ttcaagctgt
     1081 attttcagaa ccctcagttc atgggcggct gggacaagga taaggagaca gactatcggg
     1141 ccaccatcct gagatacggc tccaagtact atctggccat catggataag aagtacgcca
     1201 agtgcctgca gaagatcgac aaggacgatg tgaacggcaa ttacgagaag atcaactata
     1261 agctgctgcc cggccctaat aagatgctgc caaaggtgtt cttttctaag aagtggatgg
     1321 cctactataa ccccagcgag gacatccaga agatctacaa gaatggcaca ttcaagaagg
     1381 gcgatatgtt taacctgaat gactgtcaca agctgatcga cttctttaag gatagcatct
     1441 cccggtatcc aaagtggtcc aatgcctacg atttcaactt ttctgagaca gagaagtata
     1501 aggacatcgc cggcttttac agagaggtgg aggagcaggg ctataaggtg agcttcgagt
     1561 ctgccagcaa gaaggaggtg gataagctgg tggaggaggg caagctgtat atgttccaga
     1621 tctataacaa ggacttttcc gataagtctc acggcacacc caatctgcac accatgtact
     1681 tcaagctgct gtttgacgag aacaatcacg gacagatcag gctgagcgga ggagcagagc
     1741 tgttcatgag gcgcgcctcc ctgaagaagg aggagctggt ggtgcaccca gccaactccc
     1801 ctatcgccaa caagaatcca gataatccca agaaaaccac aaccctgtcc tacgacgtgt
     1861 ataaggataa gaggttttct gaggaccagt acgagctgca catcccaatc gccatcaata
     1921 agtgccccaa gaacatcttc aagatcaata cagaggtgcg cgtgctgctg aagcacgacg
     1981 ataaccccta tgtgatcggc atcgccaggg gcgagcgcaa tctgctgtat atcgtggtgg
     2041 tggacggcaa gggcaacatc gtggagcagt attccctgaa cgagatcatc aacaacttca
     2101 acggcatcag gatcaagaca gattaccact ctctgctgga caagaaggag aaggagaggt
     2161 tcgaggcccg ccagaactgg acctccatcg agaatatcaa ggagctgaag gccggctata
     2221 tctctcaggt ggtgcacaag atctgcgagc tggtggagaa gtacgatgcc gtgatcgccc
     2281 tggaggacct gaactctggc tttaagaata gccgcgtgaa ggtggagaag caggtgtatc
     2341 agaagttcga gaagatgctg atcgataagc tgaactacat ggtggacaag aagtctaatc
     2401 cttgtgcaac aggcggcgcc ctgaagggct atcagatcac caataagttc gagagcttta
     2461 agtccatgtc tacccagaac ggcttcatct tttacatccc tgcctggctg acatccaaga
     2521 tcgatccatc taccggcttt gtgaacctgc tgaaaaccaa gtataccagc atcgccgatt
     2581 ccaagaagtt catcagctcc tttgacagga tcatgtacgt gcccgaggag gatctgttcg
     2641 agtttgccct ggactataag aacttctctc gcacagacgc cgattacatc aagaagtgga
     2701 agctgtactc ctacggcaac cggatcagaa tcttccggaa tcctaagaag aacaacgtgt
     2761 tcgactggga ggaggtgtgc ctgaccagcg cctataagga gctgttcaac aagtacggca
     2821 tcaattatca gcagggcgat atcagagccc tgctgtgcga gcagtccgac aaggccttct
     2881 actctagctt tatggccctg atgagcctga tgctgcagat gcggaacagc atcacaggcc
     2941 gcaccgacgt ggattttctg atcagccctg tgaagaactc cgacggcatc ttctacgata
     3001 gccggaacta tgaggcccag gagaatgcca tcctgccaaa gaacgccgac gccaatggcg
     3061 cctataacat cgccagaaag gtgctgtggg ccatcggcca gttcaagaag gccgaggacg
     3121 agaagctgga taaggtgaag atcgccatct ctaacaagga gtggctggag tacgcccaga
     3181 ccagcgtgaa gcacggatcc tatccctatg acgtgcccga ttatgccagc ctgggctgtc
     3241 tgtcatacga aactgagata ctgacggtgg aatatggtct ccttcctatt gggaaaatag
     3301 tggaaaagag gatcgagtgt acggtttact cagtggacaa caatgggaat atctatacgc
     3361 aacctgtcgc tcagtggcat gatcgagggg aacaggaggt gtttgagtac tgtttggagg
     3421 acggttctct tattcgggct actaaggacc acaagtttat gacagttgat ggtcagatgt
     3481 tgcccatcga cgagatattt gagcgggaac tcgatttgat gcgggttgat aatctgccga
     3541 attaactcga gagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     3601 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3661 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3721 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3781 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3841 gtcgaggcgg ccgctgatca caattgtcta gttaaaagtt ttgttacttt atagaagaaa
     3901 ttttgagttt ttgttttttt ttaataaata aataaacata aataaattgt ttgttgaatt
     3961 tattattagt atgtaagtgt aaatataata aaacttaata tctattcaaa ttaataaata
     4021 aacctcgata tacagaccga taaaacacat gcgtcaattt tacgcatgat tatctttaac
     4081 gtacgtcaca atatgattat ctttctaggg ttaatctagt atacgcgtat gcggccgctt
     4141 aattaatcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     4201 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4261 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4321 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4381 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4441 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4501 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4561 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5041 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5281 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5341 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5401 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5461 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5521 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5581 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5641 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5701 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5761 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5821 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5881 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5941 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6001 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6061 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6121 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6181 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6241 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6301 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     6361 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     6421 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6481 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     6541 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     6601 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     6661 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     6721 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6781 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     6841 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6901 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6961 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggggcgcgcc attctagatt
     7021 aaccctagaa agatagtctg cgtaaaattg acgcatgcat tcttgaaata ttgctctctc
     7081 tttctaaata gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc
     7141 tcccgtgagg cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta taacgaccgc
     7201 gtgagtcaaa atgacgcatg attatctttt acgtgacttt taagatttaa ctcatacgat
     7261 aattatattg ttatttcatg ttctacttac gtgataactt attatatata tattttcttg
     7321 ttatagatat ca
//
