LOCUS       sequence-239376-        7886 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase (Fluc)-based bioluminescent reporter for Cre-loxP
            DNA recombination reaction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7886)
  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 7886)
  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_Floxed-STOP Fluc
FEATURES             Location/Qualifiers
     source          1..7886
                     /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"
     protein_bind    complement(917..950)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1100..1234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1354..1488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1608..1742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1759..1792)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      1799..1808
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3477..3494)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3483..3707
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3753..4181
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3840..3859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4050..4071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4190..4210)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4195..4524
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4375..4510
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4437..4456
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4591..5385
                     /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(4645..4664)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5255..5274
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5559..5680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5729..5745)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5729..5745)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5742..5764)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5753..5769
                     /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(5777..5783,5784..5801,5802..5807))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5822..5843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5960..5977)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6131..6719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6211..6230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6890..7681,7682..7750))
                     /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     7513..7532
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7751..7855)
                     /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 aagcttataa cttcgtatag catacattat acgaagttat ggtaccgcag
      961 atctgcgact ctagaggatc tgcgactcta gaggatcata atcagccata ccacatttgt
     1021 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     1081 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     1141 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1201 caaactcatc aatgtatctt atcatgtctg gatctgcgac tctagaggat cataatcagc
     1261 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1321 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     1381 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1441 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg cgactctaga
     1501 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1561 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1621 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1681 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     1741 tccccatcaa gcatcgatat aacttcgtat agcatacatt atacgaagtt atgaattcgc
     1801 cgccatggaa gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga
     1861 cgggaccgcc ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac
     1921 catcgccttt accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat
     1981 gagcgttcgg ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt
     2041 ggtgtgcagc gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg
     2101 tgtggctgtg gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg
     2161 catcagccag cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt
     2221 gcaaaagaag ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca
     2281 gggcttccaa agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta
     2341 cgacttcgtg cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag
     2401 tggcagtacc ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt
     2461 cagtcatgcc cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag
     2521 cgtggtgcca tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg
     2581 ctttcgggtc gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga
     2641 ctataagatt caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac
     2701 tctcatcgac aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct
     2761 cagcaaggag gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg
     2821 ctacggcctg acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc
     2881 tggcgcagta ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg
     2941 taagacactg ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag
     3001 cggctacgtt aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca
     3061 cagcggcgac atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa
     3121 gagcctgatc aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct
     3181 gcaacacccc aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga
     3241 gctgcccgcc gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt
     3301 ggactatgtg gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt
     3361 ggacgaggtg cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct
     3421 cattaaggcc aagaagtaac tcgagtctag agggcccgtt taaacccgct gatcagcctc
     3481 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3541 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3601 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3661 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3721 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3781 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3841 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3901 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3961 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4021 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4081 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4141 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4201 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4261 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4321 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4381 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4441 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4501 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4561 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4621 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4681 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4741 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     4801 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4861 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4921 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4981 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5041 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5101 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5161 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5221 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5281 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5341 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5401 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5461 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5521 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5581 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5641 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5701 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5761 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5821 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5881 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5941 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6001 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6061 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6121 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6181 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6241 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6301 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6361 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6421 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6481 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6541 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6601 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6661 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6721 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6781 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6841 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6901 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6961 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7021 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7081 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7141 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7201 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7261 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7321 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7381 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7441 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7501 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7561 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7621 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7681 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7741 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7801 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7861 ttccccgaaa agtgccacct gacgtc
//
