LOCUS       sequence-329179-        4663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty constitutive expression vector with piggyBac, IRES, and 
            mClover3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4663)
  AUTHORS   Chu D, Nguyen A, Smith SS, Vavrusova Z, Schneider RA
  TITLE     Stable integration of an optimized inducible promoter system enables
            spatiotemporal control of gene expression throughout avian 
            development.
  JOURNAL   Biol Open. 2020 Oct 6;9(10). pii: bio.055343. doi: 
            10.1242/bio.055343.
  PUBMED    32917762
REFERENCE   2  (bases 1 to 4663)
  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: "Biol 
            Open."; date: "2020-10-6"; volume: "9(10). pii: bio.055343. doi"; 
            pages: " 10.1242/bio.055343"
COMMENT     Sequence Label: pEPIC1.1
FEATURES             Location/Qualifiers
     source          1..4663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   5..39
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        324..823
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(359..382)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     756..775
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             866..931
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(941..946,947..970,971..1006,1007..1030)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     misc_feature    1079..1630
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1223..1240)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1450..1469
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1642..1644,1645..1647,1648..2361)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1687..1708)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2295..2316
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2425..2444)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2490..2545
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2544..2563)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   3048..3110
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        3122..3185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3186..3254,3255..4043)
                     /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(3404..3423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4072..4660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4561..4580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc
       61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga
      121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacaacc
      181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg
      241 ataattatat tgttatttcg tgttctactt acgtgataac ttattatata tatattttct
      301 tgttatagat atcagggaca gccgggtagg ggaggcgctt ttcccaaggc agtctggagc
      361 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac
      421 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc
      481 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
      541 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
      601 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
      661 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
      721 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
      781 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgattgccg ccaccttaag
      841 gtggacatct cgaggttcag caggtgatta caaggaccat gatggcgatt ataaggatca
      901 cgacatcgac tacaaagatg acgatgacaa gggtgccagc agcgcatggt cccaccctca
      961 gtttgagaag ggcggaggct ctggcggcgg aagcggagga tctgcttgga gccaccccca
     1021 gtttgaaaag taatgagaat tcgactggtt cacactatat ctgctctacc cccccctaac
     1081 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     1141 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     1201 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     1261 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     1321 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     1381 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     1441 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     1501 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     1561 aggttaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     1621 gatgataata tggccacaac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1681 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt ccgcggcgag
     1741 ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag
     1801 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcgtggc ctgcttcagc
     1861 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1921 gtccaggagc gcaccatctc tttcaaggac gacggtacct acaagacccg cgccgaggtg
     1981 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2041 gacggcaaca tcctggggca caagctggag tacaacttca acagccacta cgtctatatc
     2101 acggccgaca agcagaagaa ctgcatcaag gctaacttca agatccgcca caacgttgag
     2161 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2221 gtgctgctgc ccgacaacca ctacctgagc catcagtcca agctgagcaa agaccccaac
     2281 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccggaat aacgcacggc
     2341 atggacgagc tgtacaagtg aagcggccgg ccgccagcac agtggtcgat cgaccaatgc
     2401 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     2461 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     2521 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     2581 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     2641 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     2701 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     2761 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     2821 cctctcctga ctactcccag tcatagctgt ccctcttctc ggctgtccct cataaaagtt
     2881 ttgttacttt atagaagaaa ttttgagttt ttgttttttt ttaataaata aataaacata
     2941 aataaattgt ttgttgaatt tattattagt atgtaagtgt aaatataata aaacttaata
     3001 tctattcaaa ttaataaata aacctcgata tacagaccga taaaacacat gcgtcaattt
     3061 tacgcatgat tatctttaac gtacgtcaca atatgattat ctttctaggg ttaacgagaa
     3121 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3181 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3241 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3301 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3361 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3421 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3481 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3541 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3601 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3661 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3721 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3781 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3841 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3901 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3961 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4021 gtgcctcact gattaagcat tggtacccgt agaaaagatc aaaggatctt cttgagatcc
     4081 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4141 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4201 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4261 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4321 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4381 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4441 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4501 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4561 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4621 atttttgtga tgctcgtcag gggggcggag cctatggaaa aac
//
