LOCUS       sequence-241061-        5855 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 5855)
  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 5855)
  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: Antares2-N1
FEATURES             Location/Qualifiers
     source          1..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      571..580
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1261..1767
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="FTLEDFVGDWRQTAGYNLSQVLEQGGVSSLFQNLGVSVTPIQRIV
                     LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDNHHFKVILHYGTLVIDGVTP
                     NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL
                     HERILA"
     polyA_signal    2583..2704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2620..2639)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2674..2693
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2711..3166)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2848..2869)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3060..3079
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3193..3297
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3299..3656
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3299..3490
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3322..3342)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3507..3642
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3691..4485
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3691..4485
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3745..3764)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4355..4374
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4673..4692)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4717..4764
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5093..5681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5582..5601
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttgcc gccaccatgg tgagcaaggg cgaggagctg
      601 atcaaggaga acatgagaag caagctgtac ctggaaggca gcgtgaacgg ccaccagttc
      661 aagtgcaccc acgaagggga gggcaagccc tacgagggca agcagaccaa caggatcaag
      721 gtggtggagg gaggccccct gccgttcgca ttcgacatcc tggccaccca ctttatgtac
      781 gggagcaagg tgttcatcaa gtaccccgcc gacctccccg attattttaa gcagtccttc
      841 cctgagggct tcacatggga gagagtcatg gtgttcgaag acgggggcgt gctgaccgcc
      901 acccaggaca ccagcctcca ggacggcgag ctcatctaca acgtcaaggt cagaggggtg
      961 aacttcccag ccaacggccc cgtgatgcag aagaaaacac tgggctggga gcccagcacc
     1021 gagaccatgt accccgctga cggcggcctg gaaggcagat gcgacaaggc cctgaagctc
     1081 gtgggcgggg gccacctgca cgtcaacttc aagaccacat acaagtccaa gaaacccgtg
     1141 aagatgcccg gcgtccacta cgtggaccgc agactggaaa gaatcaagga ggccgacaac
     1201 gagacctacg tcgagcagta cgagcacgct gtggccagat actccaacct gggcggaggc
     1261 ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa cttgagtcaa
     1321 gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg
     1381 atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc
     1441 ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg
     1501 taccctgtgg ataatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac
     1561 ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc
     1621 gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag
     1681 cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc
     1741 tggcgtctgc atgaacgcat tctggcgagg cacgagctga tcaaggagaa catgagaagc
     1801 aagctgtacc tggaaggcag cgtgaacggc caccagttca agtgcaccca cgaaggggag
     1861 ggcaagccct acgagggcaa gcagaccaac aggatcaagg tggtggaggg aggccccctg
     1921 ccgttcgcat tcgacatcct ggccacccac tttatgtacg ggagcaaggt gttcatcaag
     1981 taccccgccg acctccccga ttattttaag cagtccttcc ctgagggctt cacatgggag
     2041 agagtcatgg tgttcgaaga cgggggcgtg ctgaccgcca cccaggacac cagcctccag
     2101 gacggcgagc tcatctacaa cgtcaaggtc agaggggtga acttcccagc caacggcccc
     2161 gtgatgcaga agaaaacact gggctgggag cccagcaccg agaccatgta ccccgctgac
     2221 ggcggcctgg aaggcagatg cgacaaggcc ctgaagctcg tgggcggggg ccacctgcac
     2281 gtcaacttca agaccacata caagtccaag aaacccgtga agatgcccgg cgtccactac
     2341 gtggaccgca gactggaaag aatcaaggag gccgacaacg agacctacgt cgagcagtac
     2401 gagcacgctg tggccagata ctccaacctg ggcggaggca tggacgagct gtacaagtaa
     2461 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2521 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2581 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2641 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2701 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2761 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2821 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2881 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     2941 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3001 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3061 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3121 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3181 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3241 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     3301 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3361 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3421 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3481 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3541 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3601 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     3661 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3721 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3781 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3841 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3901 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3961 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4021 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4081 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4141 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4201 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4261 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4321 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4381 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4441 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4501 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4561 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4621 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     4681 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4741 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4801 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4861 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4921 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     4981 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5041 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5101 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5161 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5221 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5281 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5341 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5401 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5461 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5521 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5581 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5641 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5701 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5761 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     5821 gggtcattag ttcatagccc atatatggag ttccg
//
