LOCUS       sequence-246746-        6086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid that promotes integration of the NanoLuc coding 
            sequence at the apobb.1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6086)
  AUTHORS   Thierer JH, Ekker SC, Farber SA
  TITLE     The LipoGlo reporter system for sensitive and specific monitoring of
            atherogenic lipoproteins.
  JOURNAL   Nat Commun. 2019 Jul 31;10(1):3426. doi: 10.1038/s41467-019-11259-w.
  PUBMED    31366908
REFERENCE   2  (bases 1 to 6086)
  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: "Nat 
            Commun."; date: "2019-07-31"; volume: "10(1):3426. doi"; pages: " 
            10.1038/s41467-019-11259-w"
COMMENT     Sequence Label: pDestTol2pA2-LA-nanoLuc-RA
FEATURES             Location/Qualifiers
     source          1..6086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(514..525,526..688)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1657..1681
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1684..2199
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     protein_bind    complement(2200..2224)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2983..3003)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(3011..3027)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3043..3177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3067..3086)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3121..3140
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(3616..3803,3804..3815)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3849..3865)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3849..3865)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3902..3922)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3902..3920)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3941..3957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3954..3976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3965..3981
                     /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(3989..3995,3996..4013,4014..4019))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4034..4055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4172..4189)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4343..4931)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4423..4442)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5102..5893,5894..5962))
                     /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     5725..5744
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5963..6067)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 aggctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcataca aggttttatc ctgctttaga actgtttgta
      781 tttaattatc aaactataag acagacaatc taatgccagt acacgctact caaagttgta
      841 aaacctcaga tttaacttca gtagaagctg attctcaaaa ttgttagtgt caagcctagc
      901 tcttttgggg ctgaaaagca atcctgcagt gctgaaaagc ctctcacagg cagccgatgc
      961 gggaagaggt gtattagtct tgatagagag gctgcaaata gcaggaaacg tgagcagaga
     1021 ctccctggtg tctgaaacac aggccagatg ggccctcgag caggaaacag ctatgaccat
     1081 gattacgcca agctatcaac tttgtataga aaagttgcgc tgcctggaat gaatgaagct
     1141 acactggctg aaatttgcaa gaaaatccaa ttagttgttg tagaaatgct gaggaatttt
     1201 ggtaacaact tagaggtata cttctctccc atcatggata actttaatac aattcaattg
     1261 acatttccca atggaaaggt catgacagta gctgagttcc aacaaaatgt gcaaagtatc
     1321 ctgagaagca acctagttat gatagcagat gcaatgaagc aaatcgaaag tcctgatgta
     1381 gttcttaaga aacttggaca aactttacaa gaggttgttg aaaaaggaca agaatttgtg
     1441 gacaaaatga atttaagttt attagaagat attgccgcgg ccattaacac gttctaccag
     1501 gagctgatga agatcataga agacattagt gaggcagtca tctttggttt ttcaatccct
     1561 gcaattaaaa tatatgaaat ttctcaaaac atagggaaag tgttgaatgc aaataatgga
     1621 atacatcaat ttgagcttcc tctcccattt ttccagacaa gtttgtacaa aaaagcaggc
     1681 ttgatggtct tcacactcga agatttcgtt ggggactggc gacagacagc cggctacaac
     1741 ctggaccaag tccttgaaca gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc
     1801 gtaactccga tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat
     1861 gtcatcatcc cgtatgaagg tctgagcggc gaccaaatgg gccagatcga aaaaattttt
     1921 aaggtggtgt accctgtgga tgatcatcac tttaaggtga tcctgcacta tggcacactg
     1981 gtaatcgacg gggttacgcc gaacatgatc gactatttcg gacggccgta tgaaggcatc
     2041 gccgtgttcg acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt
     2101 atcgacgagc gcctgatcaa ccccgacggc tccctgctgt tccgagtaac catcaacgga
     2161 gtgaccggct ggcggctgtg cgaacgcatt ctggcgtaaa cccagctttc ttgtacaaag
     2221 tggaatatgg tcattctgaa cagaaagtaa attaagtaaa cactaccatc ttttgaaatg
     2281 aacattcttg gacatcgaag ccaatggaca cttcactgaa gtatagtaca aacatgtata
     2341 taaagcaatt actgactcaa aacatgtctc attgttatgc tgctctatta ttctatttgt
     2401 ataaatgagg attgaaatac taagctagtt atcaaaaatg tcaactgcaa cctattgtca
     2461 tccttgttct gtatccaaca ttaatggcat gatggatggg gatcaataaa gaacttttat
     2521 tatgcaacac gacccaatct tgtcatttct atattctaga agtgttttaa actagtttgg
     2581 accattcaat aatataatat aattgaatat acaggtgtaa taacatgcaa caaaatgaaa
     2641 aataagcagt ggtacatggg tacattttag ttttttatgc aaactactgg tgactttaaa
     2701 aaaaaaatta caaataaaat attttattct atttttttca ttaggaagaa aaaagtttat
     2761 cagaactttt gtagaggtaa aaaaaactac taacatttta ctcaatccca tacctgaaaa
     2821 atctagtaaa tcaattgctt tgattagtgg ttattgaacc ttattttaac cagcttcagg
     2881 ttaacacatg ttaactactc ctttttgaca tataaactga aaatcattta atcttcatta
     2941 aaaatatttt ggatttgctt acaaactgat gtctgcctta cccaacttta ttatacatag
     3001 ttgataattc actggccgtc gttttacggt accatcgatg atgatccaga catgataaga
     3061 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3121 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3181 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3241 agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga tcagatctgc
     3301 gaagatacgg ccacgggtgc tcttgatcct gtggctgatt ttggactgtg ctgctcgcag
     3361 ctgctgatga atcacatact tcctccattt tcttccactg attgactgtt ataatttccc
     3421 taatttccag gtcaaggtgc tgtgcattgt ggtaatagat gtgacatgac gtcacttcca
     3481 aaggaccaat gaacatgtct gaccaatttc atataatgtg aaaacgattt tcataggcag
     3541 aataaataac atttaaatta aactgggcat cagcgcaatt caattggttt ggtaatagca
     3601 agggaaaata gaatgaagtg atctccaaaa aataagtact ttttgactgt aaataaaatt
     3661 gtaaggagta aaaagtactt ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat
     3721 tgatttttaa ttgtactcaa gtaaagtaaa aatccccaaa aataatactt aagtacagta
     3781 atcaagtaaa attactcaag tactttacac ctctggttct tgacccccta ccttcagcaa
     3841 gcccagcaga tccactagtt ctagagcggc cgccaccgcg gtggagctcc agcttttgtt
     3901 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3961 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4021 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4081 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4141 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4201 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4261 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4321 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4381 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4441 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4501 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4561 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4621 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4681 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4741 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4801 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4861 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4921 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4981 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5041 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5101 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5161 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5221 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5281 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5341 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5401 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5461 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5521 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5581 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5641 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5701 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5761 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5821 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5881 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5941 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6001 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6061 ttccgcgcac atttccccga aaagtg
//
