LOCUS       sequence-239363-        7406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Catalytically inactive mutant of PA-Cre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  AUTHORS   Kawano F, Okazaki R, Yazawa M, Sato M
  TITLE     A photoactivatable Cre-loxP recombination system for optogenetic 
            genome engineering.
  JOURNAL   Nat Chem Biol. 2016 Dec;12(12):1059-1064. doi: 
            10.1038/nchembio.2205. Epub 2016 Oct 10.
  PUBMED    27723747
REFERENCE   2  (bases 1 to 7406)
  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"; doi: 
            "10.1038/nchembio.2205"; journalName: "Nat Chem Biol"; date: 
            "2016-12"; volume: "12"; issue: "12"; pages: "1059-1064"
COMMENT     Sequence Label: pcDNA3.1_PA-Cre-Y324F
FEATURES             Location/Qualifiers
     source          1..7406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1526..1546
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1562..1618
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1628..1648
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2997..3014)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3003..3227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3273..3701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3360..3379)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3570..3591
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3710..3730)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3715..4044
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3895..4030
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3957..3976
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4111..4905
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4165..4184)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4775..4794
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5079..5200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5116..5135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5170..5189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5249..5265)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5249..5265)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5262..5284)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5273..5289
                     /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(5297..5303,5304..5321,5322..5327))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5342..5363
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5480..5497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5651..6239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5731..5750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6410..7201,7202..7270))
                     /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     7033..7052
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7271..7375)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttacca ccatggccac ctctgatgaa gtcaggaaga acctgatgga
      961 catgttcagg gacaggcagg ccttctctga acacacctgg aagatgctcc tgtctgtgtg
     1021 cagatcctgg gctgcctggt gcaagctgaa cggtacccat actctttatg cccccggtgg
     1081 atatgacatt atgggatatc tggaccagat cggcaaccgg ccaaacccgc aggtggaact
     1141 gggccccgtg gatacatcct gcgccttgat tctttgtgac ctgaaacaga aagacacccc
     1201 gatagtttac gcgagtgaag ccttcctcta catgacaggt tacagcaacg cagaggtgct
     1261 gggccggaat tgccggtttc tgcaaagccc tgacggcatg gtgaagccca agagcacccg
     1321 gaagtacgtg gatagtaaca caatcaatac tatgcgcaag gcaatcgaca ggaatgccga
     1381 ggtgcaggtt gaagtagtca attttaaaaa gaatggacag cgatttgtta atttcctgac
     1441 tatgatacct gttagggacg aaacaggcga gtatcgatac tctatgggat tccagtgcga
     1501 aacagaaggc ggaagcggtg gcgtgcccaa gaagaagagg aaagtcggat ccggcagcgg
     1561 cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc ccggccccct
     1621 cgaggtgccc aagaagaaga ggaaagtcgg cggacatact ctttatgccc ccggtggata
     1681 tgacattatg ggatatctga ggcagatcag gaaccggcca aacccgcagg tggaactggg
     1741 ccccgtggat acatcctgcg ccttgattct ttgtgacctg aaacagaaag acaccccgat
     1801 agtttacgcg agtgaagcct tcctctacat gacaggttac agcaacgcag aggtgctggg
     1861 ccggaattgc cggtttctgc aaagccctga cggcatggtg aagcccaaga gcacccggaa
     1921 gtacgtggat agtaacacaa tcaatactat gcgcaaggca atcgacagga atgccgaggt
     1981 gcaggttgaa gtagtcaatt ttaaaaagaa tggacagcga tttgttaatt tcctgactat
     2041 gatacctgtt agggacgaaa caggcgagta tcgatactct atgggattcc agtgcgaaac
     2101 agaaggtacc aacaggaaat ggttccctgc tgaacctgag gatgtgaggg actacctcct
     2161 gtacctgcaa gccagaggcc tggctgtgaa gaccatccaa cagcacctgg gccagctcaa
     2221 catgctgcac aggagatctg gcctgcctcg cccttctgac tccaatgctg tgtccctggt
     2281 gatgaggaga atcagaaagg agaatgtgga tgctggggag agagccaagc aggccctggc
     2341 ctttgaacgc actgactttg accaagtcag atccctgatg gagaactctg acagatgcca
     2401 ggacatcagg aacctggcct tcctgggcat tgcctacaac accctgctgc gcattgccga
     2461 aattgccaga atcagagtga aggacatctc ccgcaccgat ggtgggagaa tgctgatcca
     2521 cattggcagg accaagaccc tggtgtccac agctggtgtg gagaaggccc tgtccctggg
     2581 ggttaccaag ctggtggaga gatggatctc tgtgtctggt gtggctgatg accccaacaa
     2641 ctacctgttc tgccgggtca gaaagaatgg tgtggctgcc ccttctgcca cctcccaact
     2701 gtccacccgg gccctggaag ggatctttga ggccacccac cgcctgatct atggtgccaa
     2761 ggatgactct gggcagagat acctggcctg gtctggccac tctgccagag tgggtgctgc
     2821 cagggacatg gccagggctg gtgtgtccat ccctgaaatc atgcaggctg gtggctggac
     2881 caatgtgaac attgtgatga acttcatcag aaacctggac tctgagactg gggccatggt
     2941 gaggctgctc gaagatgggg actaatctag agggcccgtt taaacccgct gatcagcctc
     3001 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3061 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3121 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3181 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3241 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3301 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3361 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3421 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3481 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3541 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3601 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3661 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3721 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3781 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3901 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3961 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4021 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4081 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4141 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4201 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4261 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4321 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4381 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4441 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4501 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4561 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4621 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4681 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4741 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4801 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4861 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4921 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4981 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5041 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5101 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5161 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5221 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5281 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5341 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5401 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5461 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5521 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5581 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5641 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5701 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5761 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5821 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5881 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5941 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6001 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6061 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6121 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6181 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6241 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6301 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6361 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6421 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6481 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6541 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6601 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6661 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6721 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6781 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6841 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6901 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6961 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7021 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7081 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7141 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7201 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7261 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7321 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7381 ttccccgaaa agtgccacct gacgtc
//
