LOCUS       sequence-238584-        7470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac transposon plasmid for live animal optical imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7470)
  AUTHORS   Karlsson J, Tzeng SY, Hemmati S, Luly KM, Choi O, Rui Y, Wilson DR, 
            Kozielski KL, Quinones-Hinojosa A, Green JJ
  TITLE     Photocrosslinked Bioreducible Polymeric Nanoparticles for Enhanced 
            Systemic siRNA Delivery as Cancer Therapy.
  JOURNAL   Adv Funct Mater. 2021 Apr 22;31(17). doi: 10.1002/adfm.202009768. 
            Epub 2021 Feb 22.
  PUBMED    34650390
REFERENCE   2  (bases 1 to 7470)
  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: "Adv Funct 
            Mater."; date: "2021-04-22"; volume: "31(17). doi"; pages: " 
            10.1002/adfm.202009768. Epub 2021 Feb 22"
COMMENT     Sequence Label: PB-Antares2
FEATURES             Location/Qualifiers
     source          1..7470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   68..130
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    427..460
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        556..859
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1109..2125
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2049..2071
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2133..2152
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2205..2214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2895..3401
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="FTLEDFVGDWRQTAGYNLSQVLEQGGVSSLFQNLGVSVTPIQRIV
                     LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDNHHFKVILHYGTLVIDGVTP
                     NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL
                     HERILA"
     primer_bind     complement(4181..4200)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4246..4301
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4300..4319)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4972..5006
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5029..5045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5029..5045)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5042..5064)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5053..5069
                     /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(5077..5083,5084..5101,5102..5107))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5122..5143
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5202..5397
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5248..5383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5403..5537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5440..5459)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5494..5513
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5605..5622)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5776..6364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5856..5875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6535..7326,7327..7395))
                     /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     7158..7177
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7396..7470,1..30))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc
       61 gacttaaccc tagaaagata atcatattgt gacgtacgtt aaagataatc atgcgtaaaa
      121 ttgacgcatg tgttttatcg atctgtatat cgaggtttat ttattaattt gaatagatat
      181 taagttttat tatatttaca cttacatact aataataaat tcaacaaaca atttatttat
      241 gtttatttat ttattaaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac
      301 ttttaaacat tctctctttt acaaaaataa acttattttg tactttaaaa acagtcatgt
      361 tgtattataa aataagtaat tagcttaact tatacataat agaaacaaat tatacttatt
      421 aatcgcataa cttcgtataa tgtatgctat acgaagttat gcgattaagg gatctgtagg
      481 gcgcagtagt ccaactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      541 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      601 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      661 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      721 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      781 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      841 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      901 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      961 gggggcgggg gggggggggg gggggcgggg cggggcgrgg cggagrggkg cggcggcagc
     1021 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
     1081 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
     1141 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
     1201 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
     1261 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
     1321 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     1381 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     1441 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
     1501 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1561 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     1621 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     1681 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggrgg gctcgggggr
     1741 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1801 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     1861 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     1921 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     1981 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     2041 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     2101 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
     2161 tcattttggc aaagaattcc actataggga gacccaagct tgccgccacc atggtgagca
     2221 agggcgagga gctgatcaag gagaacatga gaagcaagct gtacctggaa ggcagcgtga
     2281 acggccacca gttcaagtgc acccacgaag gggagggcaa gccctacgag ggcaagcaga
     2341 ccaacaggat caaggtggtg gagggaggcc ccctgccgtt cgcattcgac atcctggcca
     2401 cccactttat gtacgggagc aaggtgttca tcaagtaccc cgccgacctc cccgattatt
     2461 ttaagcagtc cttccctgag ggcttcacat gggagagagt catggtgttc gaagacgggg
     2521 gcgtgctgac cgccacccag gacaccagcc tccaggacgg cgagctcatc tacaacgtca
     2581 aggtcagagg ggtgaacttc ccagccaacg gccccgtgat gcagaagaaa acactgggct
     2641 gggagcccag caccgagacc atgtaccccg ctgacggcgg cctggaaggc agatgcgaca
     2701 aggccctgaa gctcgtgggc gggggccacc tgcacgtcaa cttcaagacc acatacaagt
     2761 ccaagaaacc cgtgaagatg cccggcgtcc actacgtgga ccgcagactg gaaagaatca
     2821 aggaggccga caacgagacc tacgtcgagc agtacgagca cgctgtggcc agatactcca
     2881 acctgggcgg aggcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct
     2941 acaacttgag tcaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg
     3001 tgtccgtaac tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca
     3061 tccatgtcat catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa
     3121 tttttaaggt ggtgtaccct gtggataatc atcactttaa ggtgatcctg cactatggca
     3181 cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag
     3241 gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca
     3301 aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca
     3361 acggagtgac cggctggcgt ctgcatgaac gcattctggc gaggcacgag ctgatcaagg
     3421 agaacatgag aagcaagctg tacctggaag gcagcgtgaa cggccaccag ttcaagtgca
     3481 cccacgaagg ggagggcaag ccctacgagg gcaagcagac caacaggatc aaggtggtgg
     3541 agggaggccc cctgccgttc gcattcgaca tcctggccac ccactttatg tacgggagca
     3601 aggtgttcat caagtacccc gccgacctcc ccgattattt taagcagtcc ttccctgagg
     3661 gcttcacatg ggagagagtc atggtgttcg aagacggggg cgtgctgacc gccacccagg
     3721 acaccagcct ccaggacggc gagctcatct acaacgtcaa ggtcagaggg gtgaacttcc
     3781 cagccaacgg ccccgtgatg cagaagaaaa cactgggctg ggagcccagc accgagacca
     3841 tgtaccccgc tgacggcggc ctggaaggca gatgcgacaa ggccctgaag ctcgtgggcg
     3901 ggggccacct gcacgtcaac ttcaagacca catacaagtc caagaaaccc gtgaagatgc
     3961 ccggcgtcca ctacgtggac cgcagactgg aaagaatcaa ggaggccgac aacgagacct
     4021 acgtcgagca gtacgagcac gctgtggcca gatactccaa cctgggcgga ggcatggacg
     4081 agctgtacaa gtaaagcggc cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg
     4141 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     4201 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     4261 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     4321 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     4381 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     4441 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     4501 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     4561 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     4621 ggagatccct cgacctgcag agctccagcc tacacaatcg ctcaagacgt gtaatgcttt
     4681 tattatatat tagtcacgat atctataaca agaaaatata tatataataa gttatcacgt
     4741 aagtagaaca tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa
     4801 aagataatca tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt
     4861 accgcattga caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc
     4921 acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca
     4981 attttacgca gactatcttt ctagggttaa aagcttggcg taatcatggt catagctgtt
     5041 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5101 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5161 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa taagtcagca
     5221 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     5281 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     5341 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     5401 ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5461 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5521 cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5581 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5641 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5701 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5761 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5821 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5881 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5941 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6001 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6061 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6121 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6181 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6241 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6301 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6361 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6421 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6481 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6541 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6601 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6661 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6721 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6781 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6841 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6901 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6961 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7021 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7081 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7141 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7201 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7261 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7321 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7381 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7441 tctcatgagc ggatacatat ttgaatgtat
//
