LOCUS       sequence-329182-        6925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty dox inducible vector with piggyBac, insulators, and a DTS. The
            constitutive mClover3 is fused to H2B to allow for visualization of 
            the nucleus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6925)
  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 6925)
  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: pPIDCB2
FEATURES             Location/Qualifiers
     source          1..6925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(4..592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(84..103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(675..1466,1467..1535))
                     /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     1298..1317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1536..1606)
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   1610..1672
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    1929..2153
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2160..2855)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2435..2454)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2704..2722
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2852..2861
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2879..3329
                     /label=TRE3G BI promoter
                     /note="3rd-generation Tet-responsive bidirectional promoter
                     that can be activated by binding of Tet-On(R) 3G"
     protein_bind    2959..2977
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2995..3013
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3031..3049
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3067..3085
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3103..3121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3139..3157
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3175..3193
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(3431..3450)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3496..3551
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3550..3569)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        3867..4366
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3902..3925)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4299..4318
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4376..4753
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      4766..4775
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4772..4774,4775..4777,4778..5488)
                     /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(4817..4838)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     5425..5446
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5498..5554
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5555..6301
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    6305..6529
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6887..6921
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
       61 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      121 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      181 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      241 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      301 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      361 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      421 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      481 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      541 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      601 ttttggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
      661 gggactttcc acacctacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
      781 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
      841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
      901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
      961 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     1021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     1081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     1141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     1321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1501 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaattaac cctagaaaga
     1621 taatcatatt gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca tgtgttttat
     1681 cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt attatattta
     1741 cacttacata ctaataataa attcaacaaa caatttattt atgtttattt atttattaaa
     1801 aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa agaggggcgg aagggacgtt
     1861 aggagggagg cagggaggca gggaggcagg gaggaacgga gggagagcac gagtcatcta
     1921 gaggacagcc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
     1981 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
     2041 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
     2101 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgtcac
     2161 ttgtacagct cgtccatgcc gccggtggag tggcggccct cggagcgttc gtactgttcc
     2221 accacggtgt agtcctcgtt gtgggaggtg atgtccaact tgcggtcgac gttgtaggcg
     2281 ccgggcatct gcacgggctt cttggccttg taggtggtct tgaagtccgc caggtagcgg
     2341 ccgccgtcct tcaggcgcag ggccatctta atgtcgccct tcagcacgcc gtcctcgggg
     2401 tacaaccgct cggtggacgc ttcccagccc attgtcttct tctgcattac ggggccgtca
     2461 ggagggaagt tggtgccgcg gagcttcacc ttgtagatca gggtgccgtc ctccagggag
     2521 gtgtcctggg tcacggtcac ggcgccgccg tcctcgaagt tcatcacgcg ctcccacttg
     2581 aagccctcgg ggaaggactg cttatagtag tcggggatgt cggcggggtg cttgatgaag
     2641 gccctggagc cgtacatgaa ctgaggggac aggatgtccc aggagaaggg cagggggcca
     2701 cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg
     2761 ccctcgatct cgaactcgtg gccgttcatg gagccctcca tgtgcacctt gaaccgcatg
     2821 aactccttga tcactgcctc gcccttgctc accatggtgg cggcggtgaa ttctccaggc
     2881 gatctgacgg ttcactaaac gagctctgct tatataggcc tcccaccgta cacgccacct
     2941 cgacatactc gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta
     3001 tcagtgatag agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga
     3061 gtttactccc tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag
     3121 aacgtatcta cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta
     3181 tcagtgatag agaacgtata agctttaggc gtgtacggtg ggcgcctata aaagcagagc
     3241 tcgtttagtg aaccgtcaga tcgcctggag caattccaca acacttttgt cttataccaa
     3301 ctttccgtac cacttcctac cctcgtaaag ccgccacctt aagagtcgtc acctgcagta
     3361 caagtgaagc ggccggccgc cagcacagtg gtcgatcgac caatgccctg gctcacaaat
     3421 accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     3481 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     3541 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     3601 atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc
     3661 tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa
     3721 aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa
     3781 catccctaaa attttcctta catgttttac tagccagatt tttcctcctc tcctgactac
     3841 tcccagtcat agctgtccct cttctcgggt aggggaggcg cttttcccaa ggcagtctgg
     3901 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
     3961 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
     4021 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
     4081 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
     4141 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
     4201 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
     4261 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
     4321 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcggccg ccaccatgcc
     4381 agagccagcg aagtctgctc ccgccccgaa aaagggctcc aagaaggcgg tgactaaggc
     4441 gcagaagaaa ggcggcaaga agcgcaagcg cagccgcaag gagagctatt ccatctatgt
     4501 gtacaaggtt ctgaagcagg tccaccctga caccggcatt tcgtccaagg ccatgggcat
     4561 catgaattcg tttgtgaacg acattttcga gcgcatcgca ggtgaggctt cccgcctggc
     4621 gcattacaac aagcgctcga ccatcacctc cagggagatc cagacggccg tgcgcctgct
     4681 gctgcctggg gagttggcca agcacgccgt gtccgagggt actaaggcca tcaccaagta
     4741 caccagcgct aaggttccac cggtcgccac catggtgagc aagggcgagg agctgttcac
     4801 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     4861 ccgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac
     4921 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcgtggc
     4981 ctgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     5041 cgaaggctac gtccaggagc gcaccatctc tttcaaggac gacggtacct acaagacccg
     5101 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     5161 cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttca acagccacta
     5221 cgtctatatc acggccgaca agcagaagaa ctgcatcaag gctaacttca agatccgcca
     5281 caacgttgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     5341 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc catcagtcca agctgagcaa
     5401 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccggaat
     5461 aacgcacggc atggacgagc tgtacaaggg cagcggcgcc accaacttca gcctgctgaa
     5521 gcaggccggc gacgtggagg agaaccccgg ccccatgtct agactggaca agagcaaagt
     5581 cataaactct gctctggaat tactcaatgg agtcggtatc gaaggcctga cgacaaggaa
     5641 actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga agaacaagcg
     5701 ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc actcctgccc
     5761 cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat accgctgtgc
     5821 tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa cagagaaaca
     5881 gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct ccctggagaa
     5941 cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat tggaggaaca
     6001 ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta tgcccccact
     6061 tctgaaacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct tccttttcgg
     6121 cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg gcgggccgac
     6181 cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg acgactttga
     6241 ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc tccccgggta
     6301 acgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     6361 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     6421 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     6481 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggg gctgtctacc
     6541 actgtcgctc cctccgttcc tccctgcctc cctgcctccc tgcctccctc ctaacgtccc
     6601 ttccgcccct ctgatatcta taacaagaaa atatatatat aataagttat cacgtaagta
     6661 gaacacgaaa taacaatata attatcgtat gagttaaatc ttaaaagtca cgtaaaagat
     6721 aatcatgcgt cattttgact cacgcggttg ttatagttca aaatcagtga cacttaccgc
     6781 attgacaagc acgcctcacg ggagctccaa gcggcgactg agatgtccta aatgcacagc
     6841 gacggattcg cgctatttag aaagagagag caatatttca agaatgcatg cgtcaatttt
     6901 acgcagacta tctttctagg gttaa
//
