LOCUS       sequence-414475-        5493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Cre recombinase linked to a mScarlet fluorescent 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5493)
  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 5493)
  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-mScarlet
FEATURES             Location/Qualifiers
     source          1..5493
                     /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..3071
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2509..2527)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2777..2796
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3096..3217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3133..3152)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3187..3206
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3285..3873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3365..3384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4044..4835,4836..4904))
                     /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     4667..4686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4905..5009)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5035..5490)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5172..5193)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5384..5403
                     /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 gagcaagggc gaggcagtga
     2401 tcaaggagtt catgcggttc aaggtgcaca tggagggctc catgaacggc cacgagttcg
     2461 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     2521 tgaccaaggg tggccccctg cccttctcct gggacatcct gtcccctcag ttcatgtacg
     2581 gctccagggc cttcaccaag caccccgccg acatccccga ctactataag cagtccttcc
     2641 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgcc gtgaccgtga
     2701 cccaggacac ctccctggag gacggcaccc tgatctacaa ggtgaagctc cgcggcacca
     2761 acttccctcc tgacggcccc gtaatgcaga agaagacaat gggctgggaa gcgtccaccg
     2821 agcggttgta ccccgaggac ggcgtgctga agggcgacat taagatggcc ctgcgcctga
     2881 aggacggcgg ccgctacctg gcggacttca agaccaccta caaggccaag aagcccgtgc
     2941 agatgcccgg cgcctacaac gtcgaccgca agttggacat cacctcccac aacgaggact
     3001 acaccgtggt ggaacagtac gaacgctccg agggccgcca ctccaccggc ggcatggacg
     3061 agctgtacaa gtgaggcgcg cctattatgg tacctaactt gtttattgca gcttataatg
     3121 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3181 ctagttgtgg tttgtccaaa ctcatcaatg tatcttacat atgacatgtg agcaaaaggc
     3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3721 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3961 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4021 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4081 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4141 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4201 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4261 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4321 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4381 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4441 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4501 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4561 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4621 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4681 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4741 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4801 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4861 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4921 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4981 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     5041 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5101 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5161 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5221 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     5281 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5341 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5401 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5461 ccgcgcttaa tgcgccgcta cagggcgcgt ccc
//
