LOCUS       sequence-234378-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      631..640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1321..1827
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="FTLEDFVGDWRQTAGYNLSQVLEQGGVSSLFQNLGVSVTPIQRIV
                     LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDNHHFKVILHYGTLVIDGVTP
                     NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL
                     HERILA"
     polyA_signal    2643..2764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2680..2699)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2734..2753
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2771..3226)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2908..2929)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3120..3139
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3253..3357
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3359..3716
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3359..3550
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3382..3402)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3567..3702
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3751..4545
                     /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             3751..4545
                     /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(3805..3824)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4415..4434
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4733..4752)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4777..4824
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5153..5741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5642..5661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttgcc gccaccatgg tgagcaaggg cgaggagctg
      661 atcaaggaga acatgagaag caagctgtac ctggaaggca gcgtgaacgg ccaccagttc
      721 aagtgcaccc acgaagggga gggcaagccc tacgagggca agcagaccaa caggatcaag
      781 gtggtggagg gaggccccct gccgttcgca ttcgacatcc tggccaccca ctttatgtac
      841 gggagcaagg tgttcatcaa gtaccccgcc gacctccccg attattttaa gcagtccttc
      901 cctgagggct tcacatggga gagagtcatg gtgttcgaag acgggggcgt gctgaccgcc
      961 acccaggaca ccagcctcca ggacggcgag ctcatctaca acgtcaaggt cagaggggtg
     1021 aacttcccag ccaacggccc cgtgatgcag aagaaaacac tgggctggga gcccagcacc
     1081 gagaccatgt accccgctga cggcggcctg gaaggcagat gcgacaaggc cctgaagctc
     1141 gtgggcgggg gccacctgca cgtcaacttc aagaccacat acaagtccaa gaaacccgtg
     1201 aagatgcccg gcgtccacta cgtggaccgc agactggaaa gaatcaagga ggccgacaac
     1261 gagacctacg tcgagcagta cgagcacgct gtggccagat actccaacct gggcggaggc
     1321 ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa cttgagtcaa
     1381 gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc cgtaactccg
     1441 atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc
     1501 ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg
     1561 taccctgtgg ataatcatca ctttaaggtg atcctgcact atggcacact ggtaatcgac
     1621 ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat cgccgtgttc
     1681 gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat tatcgacgag
     1741 cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc
     1801 tggcgtctgc atgaacgcat tctggcgagg cacgagctga tcaaggagaa catgagaagc
     1861 aagctgtacc tggaaggcag cgtgaacggc caccagttca agtgcaccca cgaaggggag
     1921 ggcaagccct acgagggcaa gcagaccaac aggatcaagg tggtggaggg aggccccctg
     1981 ccgttcgcat tcgacatcct ggccacccac tttatgtacg ggagcaaggt gttcatcaag
     2041 taccccgccg acctccccga ttattttaag cagtccttcc ctgagggctt cacatgggag
     2101 agagtcatgg tgttcgaaga cgggggcgtg ctgaccgcca cccaggacac cagcctccag
     2161 gacggcgagc tcatctacaa cgtcaaggtc agaggggtga acttcccagc caacggcccc
     2221 gtgatgcaga agaaaacact gggctgggag cccagcaccg agaccatgta ccccgctgac
     2281 ggcggcctgg aaggcagatg cgacaaggcc ctgaagctcg tgggcggggg ccacctgcac
     2341 gtcaacttca agaccacata caagtccaag aaacccgtga agatgcccgg cgtccactac
     2401 gtggaccgca gactggaaag aatcaaggag gccgacaacg agacctacgt cgagcagtac
     2461 gagcacgctg tggccagata ctccaacctg ggcggaggca tggacgagct gtacaagtaa
     2521 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2581 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2641 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2701 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2761 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     2821 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     2881 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     2941 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3001 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     3061 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     3121 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     3181 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     3241 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3301 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     3361 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3421 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3481 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3541 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3601 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3661 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     3721 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3781 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3841 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3901 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     3961 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     4021 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     4081 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4141 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4201 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4261 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4321 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4381 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4441 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4501 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4561 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4621 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4681 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     4741 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4801 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     4861 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     4921 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     4981 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     5041 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5101 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5161 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5221 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5281 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5341 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5401 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5461 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5521 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5581 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5641 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5701 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5761 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5821 ctgattctgt ggataaccgt attaccgcca tgcat
//
