LOCUS       sequence-238586-        8073 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 8073)
  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 8073)
  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-Aka-P2A-GFP
FEATURES             Location/Qualifiers
     source          1..8073
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   28..90
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    387..420
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        516..819
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1098..2114
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2038..2060
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2122..2141
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2169..2188
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2169..2187
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3854..3910
                     /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"
     regulatory      3932..3941
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3938..3940,3941..3943,3944..4657)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3938..3940,3941..3943,3944..4654)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3983..4004)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4244..4263)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4591..4612
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4744..4763)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4809..4864
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4863..4882)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   5535..5569
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5592..5608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5592..5608)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5605..5627)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5616..5632
                     /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(5640..5646,5647..5664,5665..5670))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5685..5706
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5765..5960
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5811..5946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5966..6100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6003..6022)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6057..6076
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6168..6185)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6339..6927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6419..6438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7098..7889,7890..7958))
                     /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     7721..7740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7959..8063)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgaaaagtgc cacctgggtc gacttaaccc tagaaagata atcatattgt gacgtacgtt
       61 aaagataatc atgcgtaaaa ttgacgcatg tgttttatcg atctgtatat cgaggtttat
      121 ttattaattt gaatagatat taagttttat tatatttaca cttacatact aataataaat
      181 tcaacaaaca atttatttat gtttatttat ttattaaaaa aaaacaaaaa ctcaaaattt
      241 cttctataaa gtaacaaaac ttttaaacat tctctctttt acaaaaataa acttattttg
      301 tactttaaaa acagtcatgt tgtattataa aataagtaat tagcttaact tatacataat
      361 agaaacaaat tatacttatt aatcgcataa cttcgtataa tgtatgctat acgaagttat
      421 gcgattaagg gatctgtagg gcgcagtagt ccaactagtt attaatagta atcaattacg
      481 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      541 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      601 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      661 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      721 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      781 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
      841 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
      901 taattatttt gtgcagcgat gggggcgggg gggggggggg nsgcgcgccr ggsggggsgg
      961 ggsggggsgr ggggsggggs ggggsgrggc ggrgaggtgc ggcggcagcc aatcagagcg
     1021 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
     1081 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
     1141 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
     1201 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
     1261 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
     1321 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
     1381 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
     1441 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
     1501 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
     1561 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
     1621 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
     1681 gggtgccggg cggggcgggg ccgcctcggg ccggggrggg ctcgggggag gggcgcggcg
     1741 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     1801 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     1861 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     1921 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     1981 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     2041 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     2101 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     2161 aagaattcta atacgactca ctataggggc tagcatggag gacgctaaga acatcaagaa
     2221 gggcccagct ccattttatc ctctggaaga cggaacagca ggagaacagc tgcataaggc
     2281 tatgaagaga tacgctctgg tgccaggagc tattgctttt acagacgctc acattcaggt
     2341 ggacgtgaca tacgccgaat acttcgagat gagcgtgaga ctggcagaag ccatgagaag
     2401 atacggcctg aataccaacc acagaattgt ggtgtgcagc gagaacagct ctcagttctt
     2461 catgccagtg ctgggagctc tgtttattgg agtggccgtg gctccagcta acgatatcta
     2521 taacgagaga gagctgctga actctatggg aatctctcag cctacagtgg tgtttgtgag
     2581 caagaaggga ctgagaaaag tgctgaacgt gcagaagaag ctgcccatca tccggaagat
     2641 catcatcatg gacagcaaga ccgattacca gggattccag agcatgtaca ccttcgtgac
     2701 aagccatctg cctccttctt tcaacgagta cgacttcgtg ccagagtctt tcgacagaga
     2761 caagacaatc gccctgatca tgaactctag cggatctaca ggactgccta aaggagtggc
     2821 tctgcctcat agaacagctt gcgtgagatt cagccacgct agagatccta tcttcggata
     2881 ccagaacatc ccagatacag ccattctgag cgtggtgcct tttcaccacg gatttggcat
     2941 gttcacaaca ctgggctacc tgatttgcgg attcagagtg gtgctgatgt acaggttcga
     3001 ggaagaactg tttctgagaa gcctccagga ctacaagatt cagtcagctc tgctggtgcc
     3061 tacactgttt tcttgtctgg ccaagagcac actgatcgat aagtacgacc tgagcagcct
     3121 gagagaaatt gcttcaggag gagctcctct gtctaaggaa gtgggagaag cagtggctaa
     3181 gagattcaga ctgccaggaa tcagacaggg atacggactg acagaaacca caaacgcagt
     3241 gatgatcacc ccagaaggag atagaaagcc aggaagcgtg ggaaaagtgg tgcctttctt
     3301 tgaggctaag gtggtggatc tggtgacagg aaaaaccctg ggagtgaatc agagaggaga
     3361 actgtgcgtg agaggaccta tgatcatgtc aggatacgtg aacaatccag aggccacaaa
     3421 cgctctcatc gataaagacg gttggctgca tagcggagat atcgcttatt gggacgagga
     3481 cgaacacttc ttcatcgtgg acagactgaa gagcctgatc aagtacaagg gctatcaggt
     3541 ggctccagca gaactggaag gcatcctgct ccagcaccct tatatttttg acgccggagt
     3601 ggcaggactg ccagacgacg acgcaggaga actgccagca gcagtcgtgg tgctggaaca
     3661 cggaaaaacc atgaccgaga aggagatcgt ggactacgtg gcttctcagg tgacaacagc
     3721 taagaaactg agaggaggag tggtgtttgt ggacgaagtg cctagaggat ctacaggaaa
     3781 gctggacgct agaaagatca gagagatcct gaccaaggcc aagaaggacg gaaaaattgc
     3841 cgtgggaagc ggagctacta acttcagcct gctgaagcag gctggagacg tggaggagaa
     3901 ccctggacct ataacccggg atccaccggt cgccaccatg gtgagcaagg gcgaggagct
     3961 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     4021 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     4081 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     4141 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     4201 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     4261 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     4321 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     4381 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     4441 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     4501 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     4561 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     4621 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcactc ctcaggtgca
     4681 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     4741 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     4801 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     4861 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     4921 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     4981 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     5041 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     5101 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     5161 catagctgtc cctcttctct tatggagatc cctcgacctg cagagctcca gcctacacaa
     5221 tcgctcaaga cgtgtaatgc ttttattata tattagtcac gatatctata acaagaaaat
     5281 atatatataa taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga
     5341 gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt
     5401 atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc
     5461 ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca
     5521 atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt taaaagcttg
     5581 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5641 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5701 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg
     5761 atccgcatct caataagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5821 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5881 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5941 gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata
     6001 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6061 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc
     6121 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6181 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6241 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6301 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6361 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6421 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6481 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6541 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6601 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6661 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6721 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6781 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6841 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6901 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6961 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     7021 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     7081 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7141 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7201 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7261 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7321 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7381 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7441 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7501 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7561 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7621 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7681 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7741 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7801 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7861 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7921 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7981 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     8041 aataaacaaa taggggttcc gcgcacattt ccc
//
