LOCUS       sequence-466924-        7490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac transposon plasmid for live animal optical imaging of 
            Red-enhanced nanolantern.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7490)
  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 7490)
  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-ReNL
FEATURES             Location/Qualifiers
     source          1..7490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   59..121
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    418..451
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        547..850
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        852..1127
                     /label=chicken beta-actin promoter
     intron          1128..2144
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2068..2090
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2152..2171
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4192..4211)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4257..4312
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4311..4330)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   4983..5017
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5040..5056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5040..5056)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5053..5075)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5064..5080
                     /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(5088..5094,5095..5112,5113..5118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5133..5154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5213..5408
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5259..5394
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5414..5548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5451..5470)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5505..5524
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5616..5633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5787..6375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5867..5886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6546..7337,7338..7406))
                     /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     7169..7188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7407..7490,1..21))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacttaacc
       61 ctagaaagat aatcatattg tgacgtacgt taaagataat catgcgtaaa attgacgcat
      121 gtgttttatc gatctgtata tcgaggttta tttattaatt tgaatagata ttaagtttta
      181 ttatatttac acttacatac taataataaa ttcaacaaac aatttattta tgtttattta
      241 tttattaaaa aaaaacaaaa actcaaaatt tcttctataa agtaacaaaa cttttaaaca
      301 ttctctcttt tacaaaaata aacttatttt gtactttaaa aacagtcatg ttgtattata
      361 aaataagtaa ttagcttaac ttatacataa tagaaacaaa ttatacttat taatcgcata
      421 acttcgtata atgtatgcta tacgaagtta tgcgattaag ggatctgtag ggcgcagtag
      481 tccaactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      541 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      601 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      661 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      721 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      781 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      841 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
      901 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
      961 gggggggggg gsgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
     1021 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1081 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1141 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     1201 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     1261 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     1321 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     1381 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     1441 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     1501 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     1561 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     1621 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     1681 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     1741 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     1801 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     1861 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     1921 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     1981 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     2041 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     2101 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     2161 tggttattgt gctgtctcat cattttggca aagaattcat ggtgagcaag ggcgaggagg
     2221 tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt
     2281 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     2341 aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt
     2401 acggctccaa ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct
     2461 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg
     2521 tgacccagga ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca
     2581 ccaacttccc ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca
     2641 ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc
     2701 tgaaggacgg cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg
     2761 tgcaactgcc cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg
     2821 actacaccat cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctggggc
     2881 atggcaccgg cagcaccggc agcggcagct ccggcaccgc ctcctccgag gacaacaaca
     2941 tggccgtcat caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc
     3001 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     3061 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt
     3121 tcatgtacgg ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc
     3181 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg
     3241 tgaccgtgac ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc
     3301 gcggcaccaa cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     3361 cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc
     3421 tgaagctgaa ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga
     3481 agcccgtgca actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca
     3541 acgaggacta caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc
     3601 ggctggaaga tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc
     3661 ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc
     3721 aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt
     3781 atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag gtggtgtacc
     3841 ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta atcgacgggg
     3901 ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg
     3961 gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc
     4021 tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg accggctggc
     4081 ggctgtgcga acgcattctg gcgtaagcgg ccgcactcct caggtgcagg ctgcctatca
     4141 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
     4201 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     4261 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     4321 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     4381 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     4441 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     4501 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     4561 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     4621 tcttctctta tggagatccc tcgacctgca gagctccagc ctacacaatc gctcaagacg
     4681 tgtaatgctt ttattatata ttagtcacga tatctataac aagaaaatat atatataata
     4741 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
     4801 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
     4861 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
     4921 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     4981 tgcatgcgtc aattttacgc agactatctt tctagggtta aaagcttggc gtaatcatgg
     5041 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5101 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5161 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     5221 ataagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5281 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5341 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5401 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5461 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5521 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     5581 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5641 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5701 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5761 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5821 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5881 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5941 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     6001 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6061 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6121 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6181 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     6241 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     6301 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6361 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6421 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6481 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6541 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6601 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6661 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6721 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6781 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6841 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6901 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6961 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7021 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7081 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7141 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7201 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7261 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7321 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7381 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7441 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
//
