LOCUS       sequence-414477-        5508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Cre recombinase linked to a mTagBFP2 fluorescent 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5508)
  AUTHORS   Philip R, Sharma A, Matellan L, Erpf AC, Hsu WH, Tkach JM, Wyatt 
            HDM, Pelletier L
  TITLE     qTAG: an adaptable plasmid scaffold for CRISPR-based endogenous 
            tagging.
  JOURNAL   EMBO J. 2024 Dec 12. doi: 10.1038/s44318-024-00337-5.
  PUBMED    39668248
REFERENCE   2  (bases 1 to 5508)
  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: "EMBO J. 
            2024 Dec 12. doi: 10.1038/s44318-024-00337-5."
COMMENT     Sequence Label: pEF1-alpha-Cre-2A-mTagBFP2
FEATURES             Location/Qualifiers
     source          1..5508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        79..290
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          291..1229
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1186..1206
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1260..2288
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             2304..2360
                     /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             2376..3086
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    3111..3232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3148..3167)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3202..3221
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3300..3888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3380..3399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4059..4850,4851..4919))
                     /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     4682..4701
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4920..5024)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5050..5505)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5187..5208)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5399..5418
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
       61 tacgccagtc gattcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
      121 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
      181 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
      241 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
      301 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
      361 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
      421 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
      481 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
      541 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
      601 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
      661 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
      721 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
      781 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
      841 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
      901 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
      961 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     1021 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     1081 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     1141 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     1201 tggttcaaag tttttttctt ccatttcagg tgtcgtgagg atccgaattc acgcgtctaa
     1261 tgagcaattt actgaccgta caccaaaatt tgcctgcatt accggtcgat gcaacgagtg
     1321 atgaggttcg caagaacctg atggacatgt tcagggatcg ccaggcgttt tctgagcata
     1381 cctggaaaat gcttctgtcc gtttgccggt cgtgggcggc atggtgcaag ttgaataacc
     1441 ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta tcttctatat cttcaggcgc
     1501 gcggtctggc agtaaaaact atccagcaac atttgggcca gctaaacatg cttcatcgtc
     1561 ggtccgggct gccacgacca agtgacagca atgctgtttc actggttatg cggcggatcc
     1621 gaaaagaaaa cgttgatgcc ggtgaacgtg caaaacaggc tctagcgttc gaacgcactg
     1681 atttcgacca ggttcgttca ctcatggaaa atagcgatcg ctgccaggat atacgtaatc
     1741 tggcatttct ggggattgct tataacaccc tgttacgtat agccgaaatt gccaggatca
     1801 gggttaaaga tatctcacgt actgacggtg ggagaatgtt aatccatatt ggcagaacga
     1861 aaacgctggt tagcaccgca ggtgtagaga aggcacttag cctgggggta actaaactgg
     1921 tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc gaataactac ctgttttgcc
     1981 gggtcagaaa aaatggtgtt gccgcgccat ctgccaccag ccagctatca actcgcgccc
     2041 tggaagggat ttttgaagca actcatcgat tgatttacgg cgctaaggat gactctggtc
     2101 agagatacct ggcctggtct ggacacagtg cccgtgtcgg agccgcgcga gatatggccc
     2161 gcgctggagt ttcaataccg gagatcatgc aagctggtgg ctggaccaat gtaaatattg
     2221 tcatgaacta tatccgtaac ctggatagtg aaacaggggc aatggtgcgc ctgctggaag
     2281 atggcgaccc cgggggatcc ggagctacta acttcagcct gctgaagcag gccggagacg
     2341 tggaggagaa ccctggacct tctagaggaa gcggaatggt gtctaagggc gaagagctga
     2401 ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca
     2461 agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg
     2521 tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg
     2581 gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc
     2641 ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta
     2701 cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga
     2761 acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg
     2821 agacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg
     2881 tgggcgggag ccatctgatc gcaaacgcca agaccacata tagatccaag aaacccgcta
     2941 agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg
     3001 ccaacaacga gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc
     3061 ctagcaaact ggggcacaag ctgaattgag gcgcgcctat tatggtacct aacttgttta
     3121 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3181 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tacatatgac
     3241 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3301 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3361 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3421 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3481 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3541 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3601 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3661 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3721 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3781 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3841 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3901 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3961 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4021 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4081 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4141 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4201 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4261 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4321 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4381 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4441 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4501 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4561 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4621 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4681 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4741 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4801 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4861 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4921 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4981 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5041 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5101 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5161 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5221 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5281 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5341 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5401 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5461 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccc
//
