LOCUS       sequence-251298-        6086 bp    DNA     circular SYN 15-APR-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 Apr 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     613..629
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    649..669
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1209..1233
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1236..1751
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     protein_bind    complement(1752..1776)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2535..2555)
                     /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(2563..2579)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2595..2729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2619..2638)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2673..2692
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3401..3417)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3401..3417)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3454..3474)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3454..3472)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3493..3509)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3506..3528)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3517..3533
                     /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(3541..3547,3548..3565,3566..3571))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3586..3607
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3724..3741)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3895..4483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3975..3994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4654..5445,5446..5514))
                     /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     5277..5296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5515..5619)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5782..5803)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atctgttcag acacccatat ccactctgtt ccacacaggt cagaggtttg tccaggagtt
       61 cttgacagag gtgtaaaaag tactcaaaaa ttttactcaa gtgaaagtac aagtacttag
      121 ggaaaatttt actcaattaa aagtaaaagt atctggctag aatcttactt gagtaaaagt
      181 aaaaaagtac tccattaaaa ttgtacttga gtattaagga agtaaaagta aaagcaagaa
      241 agaaaactag agattcttgt ttaagctttt aatctcaaaa aacattaaat gaaatgcata
      301 caaggtttta tcctgcttta gaactgtttg tatttaatta tcaaactata agacagacaa
      361 tctaatgcca gtacacgcta ctcaaagttg taaaacctca gatttaactt cagtagaagc
      421 tgattctcaa aattgttagt gtcaagccta gctcttttgg ggctgaaaag caatcctgca
      481 gtgctgaaaa gcctctcaca ggcagccgat gcgggaagag gtgtattagt cttgatagag
      541 aggctgcaaa tagcaggaaa cgtgagcaga gactccctgg tgtctgaaac acaggccaga
      601 tgggccctcg agcaggaaac agctatgacc atgattacgc caagctatca actttgtata
      661 gaaaagttgc gctgcctgga atgaatgaag ctacactggc tgaaatttgc aagaaaatcc
      721 aattagttgt tgtagaaatg ctgaggaatt ttggtaacaa cttggaggta tacttctctc
      781 ccatcatgga taactttaat acaattcaat tgacatttcc caatggaaag gtcatgacag
      841 tagctgagtt ccaacaaaat gtgcagagta tcctgagaag caacctagtt atgatagcag
      901 atgcaatgaa gcaaatcgaa agtcctgatg tagttcttaa gaaacttgga caaactttac
      961 aagaggttgt tgaaaaagga caagagtttg tggacaaaat gaatttaagt ttattagaag
     1021 atattgccgc ggccattaac acgttctacc tggagctcat gaagatcata gaagacatta
     1081 gtgaggcagt catgtttggt ttttcaatcc ctgcaattaa aatatatgaa atgtctcaaa
     1141 acatagagaa agtgttgaac gcaaataatg gaatacatca atttgagctt cctctcccat
     1201 ttttccagac aagtttgtac aaaaaagcag gcttgatggt cttcacactc gaagatttcg
     1261 ttggggactg gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg
     1321 tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga
     1381 gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg
     1441 gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc
     1501 actttaaggt gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga
     1561 tcgactattt cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg
     1621 taacagggac cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg
     1681 gctccctgct gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca
     1741 ttctggcgta aacccagctt tcttgtacaa agtggaatat ggtcattctg aacagaaagt
     1801 aaactaagta aacactacca tattttgaaa tgaacattct tggacatcga agccaatgga
     1861 cacttcactg aagtatagta caaacatgta tataaagcaa ttactgactc aaaacatgtc
     1921 tcattgttat gctgctctat tattctattt gtataaatga ggattgaaat actaagctag
     1981 ttatcaaaaa tgccacctgc aacctattgt catccttgtt ctgtatccaa cattaatggc
     2041 atgatggatg gggatcaata aagaactttt attatgcaac acgacccaat cttgtcattt
     2101 ctatattcta gaagtgtttt acactagttt ggaccattca ataatataat ataattgaat
     2161 atacaggtgt aataacatgc aacaaaatgt aaaataagca gtggtacatg ggtacatttt
     2221 agttttttat gcaaactact ggtgacttta aaaaaaaaat tacaaataaa atattttatt
     2281 ctattttttt cattaggaag aaaaaagttt atcagaactt ttgtagaggt aaaaaaaact
     2341 actaacattt tactcaatcc catacctgaa aaatctagta aatcaattgc tttgattagt
     2401 ggttattgaa ccttatttta accagcttca ggttaacaca tgttaactac tcctttttga
     2461 catataaact gaaaatcatt taatcttcat taaaaatatt ttggatttgc ttacaaactg
     2521 atgtctgcct tacccaactt tattatacat agttgataat tcactggccg tcgttttacg
     2581 gtaccatcga tgatgatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     2641 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2701 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2761 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2821 tggctgatta tgatcctcta gatcagatct gcgaagatac ggccacgggt gctcttgatc
     2881 ctgtggctga ttttggactg tgctgctcgc agctgctgat gaatcacata cttcctccat
     2941 tttcttccac tgattgactg ttataatttc cctaatttcc aggtcaaggt gctgtgcatt
     3001 gtggtaatag atgtgacatg acgtcacttc caaaggacca atgaacatgt ctgaccaatt
     3061 tcatataatg tgaaaacgat tttcataggc agaataaata acatttaaat taaactgggc
     3121 atcagcgcaa ttcaattggt ttggtaatag caagggaaaa tagaatgaag tgatctccaa
     3181 aaaataagta ctttttgact gtaaataaaa ttgtaaggag taaaaagtac ttttttttct
     3241 aaaaaaatgt aattaagtaa aagtaaaagt attgattttt aattgtactc aagtaaagta
     3301 aaaatcccca aaaataatac ttaagtacag taatcaagta aaattactca agtactttac
     3361 acctctggtt cttgaccccc taccttcagc aagcccagca gatccactag ttctagagcg
     3421 gccgccaccg cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     3481 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3541 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3601 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3661 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3721 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3781 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3841 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3901 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3961 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4021 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4081 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4141 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4201 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4261 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4321 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4381 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4441 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4501 ttctacgggg tctgacgctc agtggaacga aaactcacgt taaggcattt tggtcatgag
     4561 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4621 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaataa gtgaggcacc
     4681 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4741 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4801 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4861 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4921 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4981 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5041 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5101 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5161 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5221 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5281 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5341 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5401 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5461 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5521 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5581 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5641 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5701 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5761 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5821 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccacgctg gctaagaact
     5881 catcagcctc cccggtccat ctacccacgt accaatgcac caattggcca caatgacggc
     5941 tactacatgg tgccattcct tcctctttat aggaatggag actacctcct gtccaacaat
     6001 gctcttggat acgagtacgc ctacctgttg gacccaggtc attgcacaac accagaaatg
     6061 ccctctgatc tgcaaaagac gtgaat
//
