LOCUS       sequence-464724-        6804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre BFP Reporter; loxP-STOP-loxP-BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6804)
  AUTHORS   Tous C, Kinstlinger IS, Rice MEL, Deng J, Wong WW
  TITLE     Multiplexing light-inducible recombinases to control cell fate, 
            Boolean logic, and cell patterning in mammalian cells.
  JOURNAL   Sci Adv. 2025 May 9;11(19):eadt1971. doi: 10.1126/sciadv.adt1971. 
            Epub 2025 May 9.
  PUBMED    40344052
REFERENCE   2  (bases 1 to 6804)
  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: "Sci Adv."; 
            date: "2025-05-9"; volume: "11(19):eadt1971. doi"; pages: " 
            10.1126/sciadv.adt1971. Epub 2025 May 9"
COMMENT     Sequence Label: IK216
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(95..114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      908..1496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1397..1416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1650..1667
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1735..1869
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1889..2023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2565..2584
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2583..2638
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2684..2703
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(2789..3490)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     protein_bind    3570..3603
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3612..3746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3926..4720)
                     /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(4037..4056)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4647..4666
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     protein_bind    4763..4796
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4844..4863)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          4871..5887
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(4925..4947)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(5888..6164)
                     /label=chicken beta-actin promoter
     enhancer        6166..6545
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6576..6680
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6681..6749,6750..6804,1..737)
                     /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"
ORIGIN
        1 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
       61 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
      121 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      181 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      241 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      301 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      361 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
      421 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
      481 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
      541 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
      601 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
      661 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      721 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
      781 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
      841 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
      901 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
      961 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1021 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     1081 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     1141 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     1201 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     1261 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     1321 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     1381 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     1441 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     1501 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     1561 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     1621 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     1681 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agcggatcca
     1741 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     1801 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     1861 aaacaagtta gctttttgca aaagcctagg cctccaaaaa agcctcctca ctacttctgg
     1921 aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca
     1981 tggggcggag aatgggcgga actgggcgga gttaggggcg ggactatggt tgctgactaa
     2041 ttgagatgcg gatccgctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gggctgcagg tcgagggatc tccataagag aagagggaca
     2281 gctatgactg ggagtagtca ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa
     2341 aattttaggg atgttaaaga aaaaaataac acaaaacaaa atataaaaaa aatctaacct
     2401 caagtcaagg cttttctatg gaataaggaa tggacagcag ggggctgttt catatactga
     2461 tgacctcttt atagccaacc tttgttcatg gcagccagca tatgggcata tgttgccaaa
     2521 ctctaaacca aatactcatt ctgatgtttt aaatgatttg ccctcccata tgtccttccg
     2581 agtgagagac acaaaaaatt ccaacacact attgcaatga aaataaattt cctttattag
     2641 ccagaagtca gatgctcaag gggcttcatg atgtccccat aatttttggc agagggaaaa
     2701 agatctcagt ggtatttgtg agccagggca ttggccacac cagccaccac cttctgatag
     2761 gcagcctgca cctgaggagt gcggccgctt aattaagctt gtgccccagt ttgctaggga
     2821 ggtcgcagta tctggccact gccacctcgt gctgctcgac gtaggtctcg ttgttggcct
     2881 ccttgattct ttccagtctg tagtccacat agtagacgcc aggcatcttg aggttcttag
     2941 cgggtttctt ggatctatat gtggtcttga tgtttgcgat cagatggctc ccgcccacga
     3001 gcttcagggc catgtcgttt ctgccttcca ggccgccgtc agcggggtac agcgtctcgg
     3061 tgaaggcctc ccagccgagt gttttcttct gcatcacagg gccgttggat gtgaagttca
     3121 cccctctgat cttgacgttg tagatgaggc agccgtcctg gaggctggtg tcctgggtag
     3181 cggtcagcac gcccccgtct tcgtatgtgg tgactctctc ccatgtgaag ccctcaggga
     3241 aggactgctt gaagaagtcg gggatgccct gggtgtggtt gatgaaggtc ttgctgccgt
     3301 agaggaagct agtagccagg atgtcgaagg cgaaggggag agggccgccc tcgaccacct
     3361 tgattctcat ggtctgggtg ccctcgtagg gcttgccttc gccctcggat gtgcacttga
     3421 agtgatggtt gtccacggtg ccctccatgt acagcttcat gtgcatgttc tccttaatca
     3481 gctcgctcat ggtggcggca ccggtggatc ccgggcccgc ggtaccgtcg actgcagaat
     3541 tcaatttaaa tcgtaccgag cgactcgaga taacttcgta taatgtatgc tatacgaagt
     3601 tattcggatt tgatccagac atgataagat acattgatga gtttggacaa accacaacta
     3661 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3721 ccattataag ctgcaataaa caagttatcg atggggtggg cgaagaactc cagcatgaga
     3781 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     3841 tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg
     3901 gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga tagaaggcga
     3961 tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca gcccattcgc
     4021 cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag cggtccgcca
     4081 cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc atgatattcg
     4141 gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga
     4201 gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat
     4261 cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt
     4321 cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca gccatgatgg
     4381 atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc acttcgccca
     4441 atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg caaggaacgc
     4501 ccgtcgtggc cagccacgat agccgcgctg cctcgtcctg cagttcattc agggcaccgg
     4561 acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg aacacggcgg
     4621 catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc tccacccaag
     4681 cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat cctcatcctg
     4741 tctcttgatc agatcaaatc cgataacttc gtataatgta tgctatacga agttatacgc
     4801 gttcgctcga attaatcaat tctttgccaa aatgatgaga cagcacaata accagcacgt
     4861 tgcccaggag ctgtaggaaa aagaagaagg catgaacatg gttagcagag gctctagagc
     4921 cgccggtcac acgccagaag ccgaaccccg ccctgccccg tcccccccga aggcagccgt
     4981 ccccccgcgg acagccccga ggctggagag ggagaagggg acggcggcgc ggcgacgcac
     5041 gaaggccctc cccgcccatt tccttcctgc cggcgccgca ccgcttcgcc ccgcgcccgc
     5101 tagagggggt gcggcggcgc ctcccagatt tcggctccgc acagatttgg gacaaaggaa
     5161 gtccctgcgc cctctcgcac gattaccata aaaggcaatg gctgcggctc gccgcgcctc
     5221 gacagccgcc ggcgctccgg gggccgccgc gcccctcccc cgagccctcc ccggcccgag
     5281 gcggccccgc cccgcccggc acccccacct gccgccaccc cccgcccggc acggcgagcc
     5341 ccgcgccacg ccccgtacgg agccccgcac ccgaagccgg gccgtgctca gcaactcggg
     5401 gaggggggtg cagggggggt tgcagcccga ccgacgcgcc cacaccccct gctcaccccc
     5461 ccacgcacac accccgcacg cagcctttgt tcccctcgca gccccccccg caccgcgggg
     5521 caccgccccc ggccgcgctc ccctcgcgca cactgcggag cgcacaaagc cccgcgccgc
     5581 gcccgcagcg ctcacagccg ccgggcagcg cggagccgca cgcggcgctc cccacgcaca
     5641 cacacacgca cgcacccccc gagccgctcc ccccgcacaa agggccctcc cggagcccct
     5701 caaggctttc acgcagccac agaaaagaaa caagccgtca ttaaaccaag cgctaattac
     5761 agcccggagg agaagggccg tcccgcccgc tcacctgtgg gagtaacgcg gtcagtcaga
     5821 gccggggcgg gcggcgcgag gcggcggcgg agcggggcac ggggcgaagg cagcgcgcag
     5881 cgactcccgc ccgccgcgcg cttcgctttt tatagggccg ccgccgccgc cgcctcgcca
     5941 taaaaggaaa ctttcggagc gcgccgctct gattggctgc cgccgcacct ctccgcctcg
     6001 ccccgccccg cccctcgccc cgccccgccc cgcctggcgc gcgccccccc ccccccccgc
     6061 ccccatcgct gcacaaaata attaaaaaat aaataaatac aaaattgggg gtggggaggg
     6121 gggggagatg gggagagtga agcagaacgt ggggctcacc tcgaccatgg taatagcgat
     6181 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     6241 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     6301 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     6361 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     6421 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     6481 atatgggcta tgaactaatg accccgtaat tgattactat taataactag tcaataatca
     6541 atgtcgaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6601 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6661 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6721 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6781 ctgaagatca gttgggtgca cgag
//
