LOCUS       sequence-214269-        7831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a zinc finger specific to the human VEGFA locus, which is 
            linked to a VPR transcriptional activation domain via a StaPL(AI) 
            module, in mammalian cells. [AI = asunaprevir inhibited]..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7831)
  AUTHORS   Jacobs CL, Badiee RK, Lin MZ
  TITLE     StaPLs: versatile genetically encoded modules for engineering 
            drug-inducible proteins.
  JOURNAL   Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z.
            Epub 2018 Jul 2.
  PUBMED    29967496
REFERENCE   2  (bases 1 to 7831)
  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"; doi: 
            "10.1038/s41592-018-0041-z"; journalName: "Nat Methods"; date: 
            "2018-07"; volume: "15"; issue: "7"; pages: "523-526"
COMMENT     Sequence Label: pCS6-ZF(VEGFA)-StaPL(AI)-YFP-VPR
FEATURES             Location/Qualifiers
     source          1..7831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..317
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        318..521
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     472..492
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     518..542
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     647..663
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     647..663
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        674..692
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     674..691
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    693..717
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      736..745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1051..1077
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1804..1806,1807..1809,1810..2520)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1849..1870)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2110..2129)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2457..2478
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2542..2691
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             3139..3510
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     CDS             3529..4098
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             4165..4185
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4189..4209
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(4248..4272)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4273..4292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4274..4292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4303..4320)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4303..4319)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4312..4334)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    4724..4858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4761..4780)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4815..4834
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4984..5438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5071..5090)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5280..5301
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             join(5725..5793,5794..6585)
                     /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     complement(5943..5962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    6687..6720
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      6798..7386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7287..7306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7540..7557
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(7631..7735)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 attagatgca tgtcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
       61 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      121 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      181 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      241 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      301 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      361 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      421 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      481 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      541 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      601 gcctccggac tctagcctag gccgcgggac ggataacaat ttcacacagg aaacagctat
      661 gaccattagg cctatttagg tgacactata gaacaagttt gtacaaaaaa gcaggctggt
      721 accggtccgg aattcgccac catgggcaaa aagaagcagc acatctgcca catccagggc
      781 tgcggcaagg tgtacggcac caccagcaac ctgaggaggc accttaggtg gcacaccggg
      841 gagaggccct tcatgtgcac ctggagctac tgcggaaaaa ggttcaccag gagcagcaac
      901 ttgcagaggc acaagaggac ccacacaggc gagaagaagt tcgcctgccc cgagtgcccc
      961 aagaggttta tgaggagcga ccatctgagc aggcacatca agacccacca gaacaagaaa
     1021 ggcgggggca gtggcgggag cgggcgcgcc tatccctacg atgtgcccga ttacgctggt
     1081 ggatccggcg ggtcagatga gatggaagag tgctctcagc acggcggatc tggaggttct
     1141 acaggctgcg tggtcatagt gggcaggatc gtcttgtccg gatccggcac tagtgcgccc
     1201 atcacggcgt acgcccagca gacgagaggc ctcctagggt gtataatcac cagcctgact
     1261 ggccgggaca aaaaccaagt ggagggtgag gtccagatca tgtcaactgc tacccaaacc
     1321 ttcctggcaa cgtgcatcaa tggggtatgc tgggcagtct accacggggc cggaacgagg
     1381 accatcgcat cacccaaggg tcctgtcatc cagatgtata ccaatgtgga ccaagacctt
     1441 gtgggctggc ccgctcctca aggttcccgc tcattgacac cctgtacctg cggctcctcg
     1501 gacctttacc tggtcacgag gcacgccgat gtcattcccg tgcgccggcg aggtgatggc
     1561 aggggtagcc tgctttcgcc ccggcccatt tcctacttga aaggctcctc ggggggtccg
     1621 ctgttgtgcc ccgcgggaca cgccgtgggc ctattcaggg ccgcggtgtg cacccgtgga
     1681 gtggctaaag cggtggactt tatccctgtg gagaacctag agacaaccat gagatccccg
     1741 gtgttcacgg acaactcctc tccaccagca gtcaccctga cgcacggggg ttcaggcggt
     1801 agtatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1861 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1921 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     1981 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     2041 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2101 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2161 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2221 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     2281 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     2341 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     2401 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     2461 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     2521 ggaggctccg gaggtagcgg cgatgcttta gacgattttg acttagatat gcttggttca
     2581 gacgcgttag acgacttcga cctagacatg ttaggctcag atgcattgga cgacttcgat
     2641 ttagatatgt tgggctccga tgccctagat gactttgatc tagatatgct aggtagtgga
     2701 ggatccggag gatcccagta tctgcccgac acagatgata gacaccgaat cgaagagaaa
     2761 cgcaagcgaa cgtatgaaac cttcaaatcg atcatgaaga aatcgccctt ctcgggtccg
     2821 accgatccca ggcccccacc gagaaggatt gcggtcccgt cccgctcgtc ggccagcgtg
     2881 ccgaagcctg cgccgcagcc ctaccccttc acgtcgagcc tgagcacaat caattatgac
     2941 gagttcccga cgatggtgtt cccctcggga caaatctcac aagcctcggc gctcgcacca
     3001 gcgcctcccc aagtccttcc gcaagcgcct gccccagcgc ctgcaccggc aatggtgtcc
     3061 gccctcgcac aggcccctgc gcccgtcccc gtgctcgcgc ctggaccgcc ccaggcggtc
     3121 gctccaccgg ctccgaagcc gacgcaggcc ggagagggaa cactctccga agcacttctt
     3181 caactccagt ttgatgacga ggatcttgga gcactccttg gaaactcgac agaccctgcg
     3241 gtgtttaccg acctcgcgtc agtagataac tccgaatttc agcagctttt gaaccagggt
     3301 atcccggtcg cgccacatac aacggagccc atgttgatgg aataccccga agcaatcacg
     3361 agacttgtga cgggagcgca gcggcctccc gatcccgcac ccgcaccttt gggggcacct
     3421 ggcctcccta acggactttt gagcggcgac gaggatttct cctccatcgc cgatatggat
     3481 ttctcagcct tgctgtcaca gatttccagc ggctctggca gcggcagccg ggattccagg
     3541 gaagggatgt ttttgccgaa gcctgaggcc ggctccgcta ttagtgacgt gtttgagggc
     3601 cgcgaggtgt gccagccaaa acgaatccgg ccatttcatc ctccaggaag tccatgggcc
     3661 aaccgcccac tccccgccag cctcgcacca acaccaaccg gtccagtaca tgagccagtc
     3721 gggtcactga ccccggcacc agtccctcag ccactggacc cagcgcccgc agtgactccc
     3781 gaggccagtc acctgttgga ggaccccgat gaagagacga gccaggctgt caaagccctt
     3841 cgggagatgg ccgatactgt gattccccag aaggaagagg ctgcaatctg tggccaaatg
     3901 gacctttccc atccgccccc aaggggccat ctggatgagc tgacaaccac acttgagtcc
     3961 atgaccgagg atctgaacct ggactcaccc ctgaccccgg aattgaacga gattctggat
     4021 accttcctga acgacgagtg cctcttgcat gccatgcata tcagcacagg actgtccatc
     4081 ttcgacacat ctctgtttgg cggatcaggc ggctccgatg agatggaaga gtgctctcag
     4141 cacggcggct ccggcggctc cgaccctaag aaaaaacgga aagttgatcc aaaaaagaag
     4201 agaaaggtat agctagagta tccctcgagg ggcccaagct tacgcgtacc cagctttctt
     4261 gtacaaagtg gtccctatag tgagtcgtat tataagctag gcactggccg tcgttttaca
     4321 acgtcgtgac tgggaaaact gctagcttgg gatctttgtg aaggaacctt acttctgtgg
     4381 tgtgacataa ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt
     4441 tttaagtgta taatgtgtta aactagctgc atatgcttgc tgcttgagag ttttgcttac
     4501 tgagtatgat ttatgaaaat attatacaca ggagctagtg attctaattg tttgtgtatt
     4561 ttagattcac agtcccaagg ctcatttcag gcccctcagt cctcacagtc tgttcatgat
     4621 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     4681 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4741 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4801 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcga
     4861 tcctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt ggctggcgta
     4921 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     4981 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5041 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5101 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5161 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5221 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag
     5281 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5341 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5401 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     5461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5521 ccaggtcttg gactggtgag aacggcttgc tcggcagctt cgatgtgtgc tggagggaga
     5581 ataaaggtct aagatgtgcg atagagggaa gtcgcattga attatgtgct gtgtagggat
     5641 cgctggtatc aaatatgtgt gcccacccct ggcatgagac aataaccctg ataaatgctt
     5701 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5761 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5821 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5881 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5941 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6001 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6061 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6121 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6181 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6241 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6301 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6361 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6421 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6481 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6541 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6601 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6661 aagatccttt ttgataatct catgccataa cttcgtataa tgtatgctat acgaagttat
     6721 ggcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6781 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6841 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6901 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6961 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7021 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7081 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7141 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7201 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7261 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7321 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7381 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7441 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7501 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7561 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7621 agagcttgca attcgcgctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7681 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7741 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     7801 aaaataggcg tattacgagg ccctttcacg c
//
