LOCUS       sequence-414488-        5238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the Cre recombinase linked to a miRFP670nano3 
            fluorescent marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5238)
  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 5238)
  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-miRFP670nano3
FEATURES             Location/Qualifiers
     source          1..5238
                     /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"
     polyA_signal    2841..2962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2878..2897)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2932..2951
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3030..3618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3110..3129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3789..4580,4581..4649))
                     /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     4412..4431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4650..4754)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4780..5235)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4917..4938)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5129..5148
                     /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 gcggaatggc aaacctggac aagatgctga
     2401 acaccaccgt gaccgaggtg cgcaagttcc tgcaagcaga cagagtgtgc gtgttcaagt
     2461 tcgaggaaga ttactccggc accgtcagcc acgaagccgt ggacgacaga tggattagca
     2521 tcctgaaaac ccaggtgcag gacagatact tcatggaaac cagaggcgag gaatacgtcc
     2581 acggcagata ccaggccatc gccgacatct acacagccaa tctggtcgag tgctacagag
     2641 acctgctgat cgagtttcag gtgcgggcca ttctggctgt ccccatcctg caaggcaaga
     2701 agctgtgggg cctgctggtg gcccaccaac tggccggccc tcgggagtgg cagacctggg
     2761 aaatcgactt cctgaaacag caagccgtgg tgatgggcat cgccatccag cagagctgag
     2821 gcgcgcctat tatggtacct aacttgttta ttgcagctta taatggttac aaataaagca
     2881 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     2941 ccaaactcat caatgtatct tacatatgac atgtgagcaa aaggccagca aaaggccagg
     3001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4681 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4741 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     4801 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4861 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4921 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4981 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     5041 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     5101 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     5161 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     5221 cgctacaggg cgcgtccc
//
