LOCUS       sequence-318357-        4297 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of 3xFLAG-NLS-VP16-FRB under the ZF1/2x6-C 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4297)
  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 4297)
  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: pJB003 ZF1/2x6-C 3xFLAG-NLS-VP16-FRB
FEATURES             Location/Qualifiers
     source          1..4297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      305..314
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             314..379
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1063..1080)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1069..1293
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1339..1767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1426..1445)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1636..1657
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1790..1986
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1837..1972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1899..1918
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2106..2227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2143..2162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2197..2216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2276..2292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2276..2292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2289..2311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2300..2316
                     /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(2324..2330,2331..2348,2349..2354))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2369..2390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2507..2524)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2678..3266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2758..2777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3437..4228,4229..4297))
                     /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     4060..4079
                     /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 tctgaattgg agtgaggacg tgtagagtga
      181 ggacgtgcgg agtgaggacg tgtagagtga ggacgtgcgg agtgaggacg tgtagagtga
      241 ggacgtgccc tctagagggt atataatggg ggccactagt ctactaccag agctcatcgc
      301 tagcgccacc atggattata aggatcacga tggcgactac aaggaccacg atattgacta
      361 caaagacgat gacgataaaa tggctcctaa gaaaaagcgc aaagtcggta tccatggcgt
      421 tcccgtcgac ggtggactcc aagccccacc aaccgatgtt tctttaggtg atgagttaca
      481 tttggacggt gaggacgtag ctatggcaca tgcagatgcc ttagacgatt ttgacttgga
      541 tatgttagga gatggagatt caccaggacc tggtttcaca ccacacgatt cagctccata
      601 cggtgctttg gacatggccg atttcgaatt tgaacagatg tttactgatg ccttgggtat
      661 agacgaatac ggtggtggag gatccggtgg tggcgggtct ggaggaggag gtagtggtgg
      721 aggtggctct ggtaccatgc ttgagatgtg gcatgaaggc ctggaagagg catctcgttt
      781 gtactttggg gaaaggaacg tgaaaggcat gtttgaggtg ctggagccct tgcatgctat
      841 gatggaacgg ggaccccaga ctctgaagga aacatccttt aatcaggcct atggtcgaga
      901 tttaatggag gcccaagagt ggtgcaggaa gtacatgaaa tcagggaatg tcaaggacct
      961 cctccaagcc tgggacctct attatcatgt gttccgacga atctcaaaga ctagatgagc
     1021 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     1081 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1141 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1201 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagtcaata
     1261 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1321 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1381 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1441 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1501 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1561 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1621 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1681 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1741 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt taggctcaat
     1801 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     1861 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     1921 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     1981 tgcaaaaagc tcccgggagc ttgtatatcc attttcggag gttgggcttc ggaatcgttt
     2041 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     2101 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2161 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2221 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     2281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2341 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2461 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2521 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2581 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2641 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2701 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2761 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2821 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2881 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2941 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3001 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3061 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3121 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3181 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3241 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3301 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3361 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3421 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3481 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3541 gcttaccatc tggccccagt gctgcaatga taccgcgaga tccacgctca ccggctccag
     3601 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3661 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3721 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3781 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3841 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3901 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3961 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4021 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4081 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4141 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4201 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4261 agggaataag ggcgacacgg aaatgttgaa tactcat
//
