LOCUS       sequence-317685-        7206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of PC 3xFLAG-FKBP-FGFR4-TEVp under the 
            EF1-alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7206)
  AUTHORS   Muldoon JJ, Kandula V, Hong M, Donahue PS, Boucher JD, Bagheri N, 
            Leonard JN
  TITLE     Model-guided design of mammalian genetic programs.
  JOURNAL   Sci Adv. 2021 Feb 19;7(8). pii: 7/8/eabe9375. doi: 
            10.1126/sciadv.abe9375. Print 2021 Feb.
  PUBMED    33608279
REFERENCE   2  (bases 1 to 7206)
  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: "2021-02-19"; volume: "7(8). pii: 7/8/eabe9375. doi"; pages: "
            10.1126/sciadv.abe9375. Print 2021 Feb"
COMMENT     Sequence Label: pJM600 EF1-alpha PC 3xFLAG-FKBP-FGFR4-TEVp in TUPV2
FEATURES             Location/Qualifiers
     source          1..7206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        486..1668
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          717..1659
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1616..1636
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1670..1694
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1853..1918
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(1919..1921,1922..2242)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     misc_feature    2304..2356
                     /label=MCS
                     /note="multiple cloning site"
     CDS             2495..3199
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="ESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKH
                     LFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQR
                     EERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIV
                     GIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEE
                     PFQPVKEATQLMN"
     primer_bind     complement(3249..3266)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3255..3479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4248..4676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4335..4354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4545..4566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4699..4895
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4746..4881
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4808..4827
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5015..5136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5052..5071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5106..5125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5185..5201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5185..5201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5198..5220)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5209..5225
                     /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)."
     promoter        complement(join(5233..5239,5240..5257,5258..5263))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5278..5299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5416..5433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5587..6175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5667..5686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6346..7137,7138..7206))
                     /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     6969..6988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtcgacg gatcgggaga tcgaagacgg gcaaaaccga tgttgacgga
      181 ctaatcctga caagatctgg ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
      241 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
      301 ctgcttcgcg atgtacgggc cagatatacg cgtgagtaat tcatacaaaa ggactcgccc
      361 ctgccttggg gaatcccagg gaccgtcgtt aaactcccac taacgtagaa cccagagatc
      421 gctgcgttcc cgccccctca cccgcccgct ctcgtcatca ctgaggtgga gaagagcatg
      481 cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
      541 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
      601 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      661 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
      721 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
      781 gaattacttc cacgcccctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
      841 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
      901 gaggcctggc ttgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
      961 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
     1021 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
     1081 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
     1141 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
     1201 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     1261 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     1321 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     1381 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     1441 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     1501 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     1561 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     1621 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgaga gctcgtttag
     1681 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagagaac
     1741 gggaccgatc caaattctcg aggctagcgc catcatgtgc cgagccatct ctcttaggcg
     1801 cttgctgctg ctgctgctgc agctgtcaca actcctagct gtcactcaag gggattacaa
     1861 ggatcacgat ggggattaca aagatcatga cattgactac aaggatgacg atgataagat
     1921 gggagtgcag gtggaaacca tctccccagg agacgggcgc accttcccca agcgcggcca
     1981 gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaatttg attcctcccg
     2041 ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga
     2101 agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta
     2161 tgcctatggt gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga
     2221 tgtggagctt ctaaaactgg aaagtgctgg tggtagtgct ggtggtagtg ctggtggtag
     2281 tgctggtggt agtgctggtg gtcctcgagc tcaagcttcg aattctgcag tcgacggtac
     2341 cgcgggcccg ggatccaccg gatctagagg cggatctgga ggctctgggg gttcaggcgg
     2401 atcaggaggc agtggcgggt caaccggtat tatactgtat gctagtggca gtctggcatt
     2461 ggcagtcctc ctgctgctcg ccgggttgga gaacgagagc ttgtttaagg ggccgcgtga
     2521 ttacaacccg atatcgagca ccatttgtca tttgacgaat gaatctgatg ggcacacaac
     2581 atcgttgtat ggtattggat ttggtccctt catcattaca aacaagcact tgtttagaag
     2641 aaataatgga acactgttgg tccaatcact acatggtgta ttcaaggtca agaacaccac
     2701 gactttgcaa caacacctca ttgatgggag ggacatgata attattcgca tgcctaagga
     2761 tttcccacca tttcctcaaa agctgaaatt tagagagcca caaagggaag agcgcatatg
     2821 tcttgtgaca accaacttcc aaactaagag catgtctagc atggtgtcag acactagttg
     2881 cacattccct tcatctgatg gcatattctg gaagcattgg attcaaacca aggatgggca
     2941 gtgtggcagt ccattagtat caactagaga tgggttcatt gttggtatac actcagcatc
     3001 gaatttcacc aacacaaaca attatttcac aagcgtgccg aaaaacttca tggaattgtt
     3061 gacaaatcag gaggcgcagc agtgggttag tggttggcga ttaaatgctg actcagtatt
     3121 gtgggggggc cataaagttt tcatggtgaa acctgaagag ccttttcagc cagttaagga
     3181 agcgactcaa ctcatgaatt agtagcggcc gctcgagtct agagggcccg tttaaacccg
     3241 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3301 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3361 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3421 caagggggag gattgggaag tcaatagcag gcatgctggg gatgcggtgg gctctatggc
     3481 ttctgaggca ggtcgaagaa ctatttcctg gctgttacgc gagcgcatcg cgataagctc
     3541 tagagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg
     3601 gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag
     3661 ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg
     3721 aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaagcttt
     3781 aggctgaaag agagatttag aatgacagaa tcatagaacg gcctgcgtac ggatggggct
     3841 ctagctctag agcttgatat cgaattcctg cagccccgct agagggacag ccccccccca
     3901 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
     3961 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
     4021 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
     4081 agcctgcaga cacctggggg gatacgggga aaaagcttta ggctgaaaga gagatttaga
     4141 atgacagaat catagaacgg cctgggcgcg cctggtggcg aacgatcaac agacgggaga
     4201 ggactaaagt cttcgctgac agcgaccgga ctctaggggg tatccccacg cgccctgtag
     4261 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4321 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4381 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4441 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4501 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4561 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4621 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4681 ctgtggaatg tgtgtcagtt aggctcaatt agtcagcaac catagtcccg cccctaactc
     4741 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4801 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4861 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4921 ttttcggagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4981 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5041 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5101 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5161 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5221 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5281 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5341 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5401 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5461 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5521 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5581 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5641 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5701 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5761 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5821 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5881 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5941 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6001 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6061 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6121 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6181 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6241 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6301 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6361 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6421 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6481 accgcgagat ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6541 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6601 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6661 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6721 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6781 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6841 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6901 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6961 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7021 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7081 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7141 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7201 actcat
//
