LOCUS       sequence-414367-        6094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of the Cre recombinase and a puro selectable marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6094)
  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 6094)
  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-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..6094
                     /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..2291
                     /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"
     polyA_signal    2310..2534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2556..3055
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2591..2614)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2988..3007
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3076..3675
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3076..3095)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3572..3592
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3697..3818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3734..3753)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3788..3807
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3886..4474)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3966..3985)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4645..5436,5437..5505))
                     /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     5268..5287
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5506..5610)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5636..6091)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5773..5794)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5985..6004
                     /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 atggcgactg accatggtct agactcgagc tgtgccttct agttgccagc catctgttgt
     2341 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2401 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2461 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2521 ggtgggctct atgggcggcc gcatagatag ctagcgggta ggggaggcgc ttttcccaag
     2581 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     2641 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     2701 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     2761 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     2821 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     2881 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     2941 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     3001 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     3061 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3121 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3181 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3241 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3301 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     3361 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     3421 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     3481 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     3541 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     3601 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     3661 aagcccggtg cctgaggcgc gcctattatg gtacctaact tgtttattgc agcttataat
     3721 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3781 tctagttgtg gtttgtccaa actcatcaat gtatcttaca tatgacatgt gagcaaaagg
     3841 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3901 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3961 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4021 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4081 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4141 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4201 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4261 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4321 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4381 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4441 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4501 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4561 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4621 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4681 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4741 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4801 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4861 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4921 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4981 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5041 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5101 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5161 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5221 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5281 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5341 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5401 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5461 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5521 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5581 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     5641 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5701 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5761 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5821 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5881 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5941 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6001 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6061 gccgcgctta atgcgccgct acagggcgcg tccc
//
