LOCUS       sequence-227444-        7419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ePiggyBac mammalian expression plasmid for tandem dimer 
            MS2cp-tagRFP-T fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7419)
  AUTHORS   Alexander JM, Guan J, Li B, Maliskova L, Song M, Shen Y, Huang B, 
            Lomvardas S, Weiner OD
  TITLE     Live-cell imaging reveals enhancer-dependent Sox2 transcription in 
            the absence of enhancer proximity.
  JOURNAL   Elife. 2019 May 24;8. pii: 41769. doi: 10.7554/eLife.41769.
  PUBMED    31124784
REFERENCE   2  (bases 1 to 7419)
  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: "Elife."; 
            date: "2019-05-24"; volume: "8. pii: 41769. doi"; pages: " 
            10.7554/eLife.41769"
COMMENT     Sequence Label: epB_UbC_tdMS2cp-tagRFP-Tx2
FEATURES             Location/Qualifiers
     source          1..7419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2237..2271
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        2578..3788
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3625..3644
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    3805..3829
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3899..3925
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             3899..3925
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3926..3937
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             4706..5437
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     CDS             5444..6175
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     CDS             6176..6196
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(6231..6255)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6371..6492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6408..6427)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6462..6481
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6678..6740
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6775..6794)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6776..6794)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6804..6821)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6804..6820)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6813..6835)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6962..7417
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7049..7068)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7259..7280
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggggcgc
     2221 gccgcagcta gattaaccct agaaagatag tctgcgtaaa attgacgcat gcattcttga
     2281 aatattgctc tctctttcta aatagcgcga atccgtcgct gtgcatttag gacatctcag
     2341 tcgccgcttg gagctcccgt gaggcgtgct tgtcaatgcg gtaagtgtca ctgattttga
     2401 actataacaa ccgcgtgagt caaaatgacg catgattatc ttttacgtga cttttaagat
     2461 ttaactcata cgataattat attgttattt cgtgttctac ttacgtgata acttattata
     2521 tatatatttt cttgttatag atatccttct tcgacattga ttattgacta gagatctggc
     2581 ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca
     2641 cgtcagacga agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg
     2701 cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac
     2761 gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa
     2821 agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat
     2881 tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc
     2941 ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc
     3001 cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa
     3061 gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag
     3121 caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt
     3181 cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg aggccttcgc
     3241 taatgcggga aagctcttat tcgggtgaga tgggctgggg caccatctgg ggaccctgac
     3301 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat
     3361 gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga
     3421 cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt
     3481 aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac
     3541 aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt
     3601 tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg
     3661 cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat
     3721 gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt
     3781 tgttagacga agctcgagga tatcacaagt ttgtacaaaa aagcaggctt taaaggaacc
     3841 aattcagtcg actggatccg gtaccgaatt cgcggccgcc tagctaacca ccatgggcta
     3901 cccctacgac gtgcccgact acgccatcga aggccgccat atgctagccg ttaaaatggc
     3961 ttctaacttt actcagttcg ttctcgtcga caatggcgga actggcgacg tgactgtcgc
     4021 cccaagcaac ttcgctaacg ggatcgctga atggatcagc tctaactcgc gttcacaggc
     4081 ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag aatcgcaaat acaccatcaa
     4141 agtcgaggtg cctaaaggcg cctggcgttc gtacttaaat atggaactaa ccattccaat
     4201 tttcgccacg aattccgact gcgagcttat tgttaaggca atgcaaggtc tcctaaaaga
     4261 tggaaacccg attccctcag caatcgcagc aaactccggc atctacgcga tctacgccat
     4321 ggcttctatg gcttctaact ttactcagtt cgttctcgtc gacaatggcg gaactggcga
     4381 cgtgactgtc gccccaagca acttcgctaa cgggatcgct gaatggatca gctctaactc
     4441 gcgttcacag gcttacaaag taacctgtag cgttcgtcag agctctgcgc agaatcgcaa
     4501 atacaccatc aaagtcgagg tgcctaaagg cgcctggcgt tcgtacttaa atatggaact
     4561 aaccattcca attttcgcca cgaattccga ctgcgagctt attgttaagg caatgcaagg
     4621 tctcctaaaa gatggaaacc cgattccctc agcaatcgca gcaaactccg gcatctacgc
     4681 ggatccaccg gtcgccaccg gatctatggt gtctaagggc gaagagctga ttaaggagaa
     4741 catgcacatg aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc
     4801 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg
     4861 cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcagaac
     4921 cttcatcaac cacacccagg gcatccccga tttctttaag cagtccttcc ctgagggctt
     4981 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     5041 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc
     5101 caacggccct gtgatgcaga agaaaacact cggctgggag gccaacaccg agatgctgta
     5161 ccccgctgac ggcggcctgg aaggcagaac cgacatggcc ctgaagctcg tgggcggggg
     5221 ccacctgatc tgcaacttca agaccacata cagatccaag aaacccgcta agaacctcaa
     5281 gatgcccggc gtctactatg tggaccacag actggaaaga atcaaggagg ccgacaaaga
     5341 gacgtacgtc gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact
     5401 ggggcacaaa cttaatggca tggacgagct gtacaagcta gggatggtgt ctaagggcga
     5461 agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca
     5521 ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag
     5581 aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt
     5641 catgtacggc agcagaacct tcatcaacca cacccagggc atccccgatt tctttaagca
     5701 gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct
     5761 gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag
     5821 aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc
     5881 caacaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaaccg acatggccct
     5941 gaagctcgtg ggcgggggcc acctgatctg caacttcaag accacataca gatccaagaa
     6001 acccgctaag aacctcaaga tgcccggcgt ctactatgtg gaccacagac tggaaagaat
     6061 caaggaggcc gacaaagaga cgtacgtcga gcagcacgag gtggctgtgg ccagatactg
     6121 cgacctccct agcaaactgg ggcacaaact taatggcatg gacgagctgt acaagcctaa
     6181 gaaaaagcgg aaagtgtaag tgcctaggcg gccgcactcg agatatctag acccagcttt
     6241 cttgtacaaa gtggtgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     6301 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     6361 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6421 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6481 caatgtatct taaggaattc gataaaagtt ttgttacttt atagaagaaa ttttgagttt
     6541 ttgttttttt ttaataaata aataaacata aataaattgt ttgttgaatt tattattagt
     6601 atgtaagtgt aaatataata aaacttaata tctattcaaa ttaataaata aacctcgata
     6661 tacagaccga taaaacacat gcgtcaattt tacgcatgat tatctttaac gtacgtcaca
     6721 atatgattat ctttctaggg ttaatctagc tgcggcgcgc cggtacccaa ttcgccctat
     6781 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     6841 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     6901 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     6961 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     7021 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     7081 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     7141 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     7201 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     7261 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     7321 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     7381 aacgcgaatt ttaacaaaat attaacgctt acaatttag
//
